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1.
Article in Chinese | WPRIM | ID: wpr-1011462

ABSTRACT

As the pace of society increases and lifestyles change, the incidence and mortality rates of breast cancer continue to rise. Targeted therapies are now promising in the treatment of breast cancer, and a variety of protein targets have been identified to play an important role in the development of breast cancer. Among them, signal transducer and activator of transcription (STAT) proteins constitute a crucial group that serves as important targets for transducing cellular transcriptional information, which can regulate downstream cell proliferation, apoptosis, cell migration, invasion, angiogenic factors, etc. and then affect the progression of breast cancer. The STAT family is closely associated with the inflammatory response to tumors and plays a landmark role in tumor development as well as in diagnosis and prognosis. The "inflammation-cancer" transformation refers to the process in which the inflammatory microenvironment caused by uncontrolled inflammation promotes normal cells to become cancerous. According to the theory of Chinese medicine, "heat toxicity" in "cancer toxicity" corresponds to inflammation, which is closely related to tumor development. As a major link associated with the inflammatory response, the STAT family has a promising role in the development and treatment of a variety of tumors, but its relevance to breast cancer remains inadequately explored. Chinese medicine has been shown to have good efficacy in the prevention and treatment of breast cancer, and some current studies have shown that the active ingredients and compounds of Chinese medicine have certain intervention effects on breast cancer-related STAT proteins, but there has not been a systematic review. In order to better sort out and summarize the studies on the effects of Chinese herbal medicines based on the STAT family interventions in breast cancer, this paper reviewed the studies on Chinese herbal medicines acting on the STAT family in recent years, aiming to provide new ideas for clinical applications in breast cancer and to provide thoughts for the development of STAT protein-based drugs.

2.
China Pharmacy ; (12): 296-303, 2024.
Article in Chinese | WPRIM | ID: wpr-1006613

ABSTRACT

OBJECTIVE To investigate the effect of berberine on ferroptosis in MG63 osteosarcoma cells and its mechanism. METHODS Using cells without drug treatment as control, the cell viability, proliferation, the related indexes of ferroptosis [nuclear proliferation associated-antigen (Ki67), mitochondrial ultrastructure, ferric ion (Fe2+), reactive oxygen species (ROS), malondialdehyde (MDA), and glutathione (GSH)], the protein expression of signal transducer and activator of transcription 3 (STAT3), tumor protein 53 (p53), and solute carrier family 7 member 11 (SLC7A11) were detected after being treated with different concentrations of berberine. Cells were transfected with p53 siRNA and then assigned to the control group, p53 siRNA group, berberine group, and p53 siRNA+berberine group to explore the role of p53 in berberine-induced ferroptosis. After 24 h incubation with 10.0 μmol/L berberine, the protein expressions of p53 and SLC7A11, the levels of mitochondrial membrane potential, GSH, and MDA content were determined. Cells were transfected with STAT3 overexpressed plasmid and then assigned to the control group, berberine group, STAT3 group, and STAT3+berberine group to explore the effect of STAT3 on the regulation of the p53/SLC7A11 pathway. After 24 h incubation with 10 μmol/L berberine, the protein expressions of p-STAT3, STAT3, p53, and SLC7A11 were detected. RESULTS Compared with the control cell, the concentrations of 2.5, 5.0 and 10.0 μmol/L berberine could reduce the cell viability and expression of Ki67, and induce the morphological changes in ferroptosis-related mitochondria, increase the levels of Fe2+, ROS and MDA, and the protein expression of p53, reduce the level of GSH, the binding activity of STAT3 with DNA, and the protein expressions of p-STAT3 and SLC7A11; the above differences were statistically significant (P< 0.05 or P<0.01). Compared with the berberine group,significantly down-regulated p53 protein expression and MDA level, up-regulated SLC7A11 protein expression, and increased mitochondrial membrane potential and GSH level were observed in the p53 siRNA+berberine group (P<0.01). Compared with the berberine group, the protein expressions of p-STAT3, STAT3, and SLC7A11 in the STAT3+berberine group were significantly increased (P<0.01), while the protein expression of p53 was significantly decreased (P<0.01). CONCLUSIONS Berberine can induce the ferroptosis of MG63 cells by mediating STAT3/p53/SLC7A11 signaling pathway.

3.
Article in Chinese | WPRIM | ID: wpr-1013342

ABSTRACT

ObjectiveTo investigate the effect of curcumin on the cycle arrest of human colon cancer HCT116 cells and decipher the possible molecular mechanism. MethodThe methyl thiazolyl tetrazolium (MTT) method was employed to examine the effects of curcumin (0, 12.5, 25, 50, 75, 100 μmol·L-1) and 5-fluorouracil (5-FU, 600 μmol·L-1) on the proliferation of HCT116 cells at different time points (24, 48, 72 h). Flow cytometry was employed to examine the cycle of HCT116 cells treated with curcumin (0, 25, 50, 75 μmol·L-1) and 5-FU. Western blot was employed to determine the expression of proteins in the Janus kinase 1 (JAK1)/signal transducer and activator of transcription 1 (STAT1) /cyclin-dependent kinase inhibitor 1A (p21) pathway in HCT116 cells. The binding of STAT1 to p21 promoter region was detected by chromatin immunoprecipitation (ChIP). Small interfering RNA (siRNA) was employed to measure the role of STAT1 in regulating the expression of p21 and that of JAK1 in regulating the activation of STAT1 by Western blot and cellular immunofluorescence, respectively. ResultCompared with the blank group, the HCT-116 cells treated with curcumin and 5-FU showed decreased viability (P<0.05), increased proportions of cells in the G0/G1 phase (P<0.05), decreased proportions of cells in the S phase and G2/M phase (P<0.05), down-regulated protein level of phosphorylated p21 (P<0.05), and up-regulated protein level of p21 (P<0.05). Compared with the curcumin group, the p21 siRNA+ curcumin group presented decreased proportion of cells in G0/G1 phase (P<0.05). Compared with the blank group, curcumin elevated the level of phosphorylated STAT1 (p-STAT1) (P<0.05). Compared with the curcumin group, the curcumin + STAT1 siRNA group showcased up-regulated protein level of p21 in HCT116 cells (P<0.05). The mechanism study showed that curcumin treatment enhanced the enrichment of STAT1 in the p21 promoter region (P<0.05) compared with the blank group. Compared with the blank group, curcumin up-regulated the level of phosphorylated JAK1 (p-JAK1) (P <0.05). Compared with the curcumin group, the curcumin + STAT1 siRNA group demonstrated up-regulated protein levels of p-STAT1 and p21 in HCT116 cells (P<0.05). ConclusionCurcumin may induce the cycle arrest of human colon cancer HCT116 cells by activating the JAK1/STAT1/p21 signaling pathway.

4.
Article in Chinese | WPRIM | ID: wpr-1012700

ABSTRACT

ObjectiveTo investigate the clinical efficacy of Qihuang Jianpi Zishen Granules in the treatment of systemic lupus erythematosus (SLE) and its effect on the signal transducer and activator of tranSCription 3/mammalian target of rapamycin (STAT3/mTOR) signaling pathway, and to decipher the possible mechanism. MethodSixty female SLE patients who met the criteria in the First Affiliated Hospital of Anhui University of Chinese Medicine from May 2022 to May 2023 were selected and randomized into a control group and an observation group (30 cases in each group). The control group was treated with prednisone acetate + hydroxychloroquine sulfate orally, and the observation group was additionally treated with Qihuang Jianpi Zishen granules. The treatment lasted for 8 weeks. The SLE disease activity (SLEDAI), TCM syndrome score, erythrocyte sedimentation rate (ESR), hypersensitive C-reactive protein (hs-CRP), immune indexes [immunoglobulin G (IgG), C3, C4, CD4+, and CD8+], interleukin (IL)-17, IL-23, interferon (IFN)-γ, 24 h urinary protein (24 h PRO), serum creatinine (SCr), and expression of proteins [STAT3, phosphorylated (p)-STAT3, mTOR protein and STAT3,mTOR mRNA] in the STAT3/mTOR signaling pathway were determined before and after treatment. In addition, the adverse reactions were recorded. ResultAfter 8 weeks of treatment, the total response rate in the observation group was 93.33% (28/30), which was higher than that (70.00%, 21/30) in the control group (χ2=4.007, P<0.05). After treatment, both groups showed declined SLEDAI, TCM syndrome score, ESR, hs-CRP, IgG, CD8+, IL-17, IL-23, IFN-γ, 24 h PRO, SCr, and expression of proteins in the STAT3/mTOR pathway (P<0.01) and elevated levels of C3, C4, and CD4+ (P<0.01). Moreover, the observation group had lower SLEDAI, TCM syndrome score, ESR, hs-CRP, IgG, CD8+, IL-17, IL-23, IFN-γ, 24 h PRO, SCr, and expression of proteins in the STAT3/mTOR pathway (P<0.05, P<0.01) and higher levels of C3, C4, and CD4+ (P<0.05, P<0.01) than the control group after treatment. Neither group showed serious adverse reactions during the treatment period. ConclusionQihuang Jianpi Zishen Granules can ameliorate the inflammatory response, reduce the disease activity, and mitigate the kidney injury in SLE by inhibiting the STAT3/mTOR signaling pathway to regulate the immune function.

5.
Chinese Critical Care Medicine ; (12): 813-817, 2023.
Article in Chinese | WPRIM | ID: wpr-992031

ABSTRACT

Objective:To explore the effect of signal transducer and activator of transcription 6 (STAT6) on ferroptosis in skeletal muscle cells in sepsis model and its potential mechanism.Methods:Twenty-four 8-week-old male specific pathogen free Kunming mice were divided into normal control group, sham group, sepsis model group and STAT6 inhibitor pretreatment group according to random number table method with 6 mice in each group. A mouse sepsis model was reproduced by cecal ligation and perforation (CLP). In the sham group, the skin of mice was sutured after exposing the cecum tissue. In the STAT6 inhibitor pretreatment group, 10 mg/kg AS1517499 was injected intraperitoneally 1 hour before model reproduction. The sham group and the model group were intraperitoneally injected with the same volume of normal saline. Mice in the normal control group did not receive any operation or drug intervention. The mice were sacrificed 24 hours after model reproduction, and the muscle tissue of hind limb was obtained under sterile condition. Hematoxylin-eosin (HE) staining was used to observe the histopathology with optical microscope, and mitochondrial morphological changes were observed by transmission electron microscopy after double staining with uranium acetate lead citrate. The ferroptosis marker proteins expressions of chitinase-3-like protein 1 (CHI3L1), cyclooxygenase-2 (COX-2), acyl-CoA synthetase long-chain family member 4 (ACSL4), ferritin heavy chain 1 (FTH1), and glutathione peroxidase 4 (GPx4) were detected by Western blotting.Results:Under the optical microscope, the morphology and structure of skeletal muscle tissues in the normal control and sham groups were normal. In the model group, the structure of skeletal muscle tissues was loose, the muscle fiber became smaller and atrophic, inflammatory cell infiltration and even muscle fiber loss were found. Compared with the model group, the structure of skeletal muscle tissues was tight and skeletal muscle atrophy was improved in the STAT6 inhibitor pretreatment group. The ultrastructure of skeletal muscle cell in the normal control and sham groups was normal under transmission electron microscope. The ultrastructure characteristics of skeletal muscle in the model group showed that cell membrane was broken and blister, mitochondria became smaller and membrane density increased, the mitochondrial crista decreased or disappeared, the mitochondrial outer membrane was broken, and the nucleus was normal in size but lacked chromatin condensation. Compared with the model group, the STAT6 inhibitor pretreatment group had a significant improvement in the ultrastructure of muscle cells. Compared with the normal control and sham groups, the protein expressions of CHI3L1, COX-2, ACSL4 and FTH1 in the muscle of the model group were significantly increased, while the protein expression of GPx4 was decreased significantly, indicating that the skeletal muscle cells in the mouse sepsis model showed characteristic mitochondrial injury and abnormal expression of ferroptosis markers. Compared with the model group, the protein expressions of CHI3LI, COX-2, ACSL4 and FTH1 in the STAT6 inhibitor pretreatment group were significantly decreased [CHI3L1 protein (CHI3L1/GAPDH): 0.70±0.08 vs. 0.97±0.09, COX-2 protein (COX-2/GAPDH): 0.61±0.03 vs. 0.83±0.03, ACSL4 protein (ACSL4/GAPDH): 0.75±0.04 vs. 1.02±0.16, FTH1 protein (FTH1/GAPDH): 0.49±0.06 vs. 0.76±0.13, all P < 0.05], while the protein expression of GPx4 was significantly increased (GPx4/GAPDH: 1.14±0.29 vs. 0.53±0.03, P < 0.05). Conclusions:Sepsis can induce ferroptosis in skeletal muscle cells of mice. STAT6 may mediate ferroptosis in mouse skeletal muscle cells by regulating the expressions of COX-2, ACSL4, FTH1 and GPx4, thereby inducing skeletal muscle cell injury in sepsis.

6.
Article in Chinese | WPRIM | ID: wpr-991069

ABSTRACT

Objective:To analyze the expression levels of urinary interleukin-6 (IL-6), signal transducer and activator of transcription 3 (STAT3) and heparin-binding protein (HBP) in urinary tract infection and its correlation with infection prognosis.Methods:The clinical data of 100 patients with urinary tract infection (urinary tract infection group) from January 2021 to December 2022 in the Affiliated Hospital of Jining Medical College were retrospectively analyzed. Among them, simple urinary tract infection was in 62 cases, and complex urinary tract infection was in 38 cases; after treatment, 25 cases were not cured, and 75 cases were cured. Another 50 healthy examinees were selected as the health control group. The level of urine IL-6 was detected by luminescence assay method, the level of urine STAT3 was detected by enzyme-linked immunosorbent assay method, and the level of urine HBP was detected by fluorescence immunochromatography method. The blood routine was detected by fully automated blood cell analyzer, and the blood cell count was recorded. The levels of serum C-reactive protein (CRP) and procalcitonin (PCT) were detected by radioimmunoassay. The correlation between urine IL-6, STAT3, HBP and blood routine inflammatory response markers was analyzed by Pearson method. The receiver operating characteristic (ROC) curve was used to evaluate the predictive effectiveness of urine IL-6, STAT3, HBP and blood routine inflammatory response markers in infection prognosis.Results:The urine IL-6, STAT3, HBP, and blood CRP, PCT, white blood cell count in urinary tract infection group were significantly higher than those in healthy control group: (33.19 ± 11.02) μg/L vs. (16.84 ± 3.57) μg/L, (66.77 ± 19.58) μg/L vs. (38.69 ± 11.04) μg/L, (151.98 ± 42.00) μg/L vs. (28.55 ± 9.16) μg/L, (12.57 ± 4.19) mg/L vs. (5.23 ± 1.80) mg/L, (0.58 ± 0.19) μg/L vs. (0.22 ± 0.07) μg/L and (9.86 ± 3.20) × 10 9/L vs. (6.44 ± 2.13) ×10 9/L, and there were statistical differences ( P<0.01). The urine IL-6, STAT3, HBP, and blood CRP, PCT, white blood cell count in patients with complex urinary tract infection were significantly higher than those in patients with simple urinary tract infection: (40.25 ± 10.34) μg/L vs. (28.87 ± 8.55) μg/L, (79.50 ± 17.92) μg/L vs. (58.96 ± 13.43) μg/L, (186.51 ± 35.92) μg/L vs. (130.82 ± 39.74) μg/L, (14.09 ± 4.18) mg/L vs. (11.64 ± 3.55) mg/L, (0.64 ± 0.20) μg/L vs. (0.55 ± 0.13) μg/L and (11.27 ± 3.08) × 10 9/L vs. (8.99 ± 2.36) × 10 9/L, and there were statistical differences ( P<0.01). The urine IL-6, STAT3, HBP, and blood CRP, PCT, white blood cell count in patients with untreated urinary tract infection were significantly higher than those in patients with cured urinary tract infection: (42.97 ± 11.51) μg/L vs. (29.93 ± 8.66) μg/L, (86.81 ± 20.35) μg/L vs. (60.09 ± 17.43) μg/L, (264.27 ± 28.76) μg/L vs. (114.55 ± 21.38) μg/L, (19.11 ± 3.28) mg/L vs. (10.39 ± 2.40) mg/L, (0.85 ± 0.14) μg/L vs. (0.49 ± 0.11) μg/L and (12.26 ± 2.77) × 10 9/L vs. (9.06 ± 2.34) ×10 9/L, and there were statistical differences ( P<0.01). Pearson correlation analysis result showed that urine IL-6, STAT3, HBP were positively correlated with blood CRP, PCT, white blood cell count ( P<0.01). The ROC curve analysis result showed that the area under curve (AUC) of urine IL-6, STAT3 and HBP in predicting the infection prognosis in patients with urinary tract infection was greater than that of blood CRP, PCT and white blood cell count; moreover, the AUC and sensitivity of the combined of urine IL-6, STAT3 and HBP in predicting the infection prognosis in patients with urinary tract infection were significantly higher than the combined of blood CRP, PCT and white blood cell count (0.937 vs. 0.898 and 96.00% vs. 76.00%), but with lower specificity (81.33% vs. 98.67%). Conclusions:Urinary tract infections can cause elevated urine IL-6, STAT3 and HBP, and the degree of elevation is related to the types of simple or complicated infection and the infection prognosis. The combined detection of the urine IL-6, STAT3 and HBP is expected to be a method to predict the infection prognosis, and it provides reference information for clinical diagnosis and treatment.

7.
China Tropical Medicine ; (12): 754-2023.
Article in Chinese | WPRIM | ID: wpr-979834

ABSTRACT

@#Abstract: Objective To investigate the influences of notoginsenoside R1 (NGR1) on cell injury and Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway of alveolar epithelial cells infected by Klebsiella pneumoniae (Kp). Methods A549 cells were grouped into five groups: control group (C group), infection group (Infect group), infection + low NGR1 group (Infect + L-NGR1 group), infection + high NGR1 group (Infect + H-NGR1 group), and infection+high NGR1+JAK2/STAT3 pathway inhibitor group (Infect+H-NGR1+SD-1029 group). Cell proliferation was measured using CCK8; ELISA kits were applied to detect the contents of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and interferon γ (IFN-γ) in the culture medium; flow cytometry was applied to detect apoptosis; RT-qPCR was applied to detect the expressions of JAK2/STAT3; Western blot was applied to detect JAK2/STAT3 pathway, autophagy protein microtubule-associated protein 1 light chain 3 (LC3), autophagy-relatedgene5 (Atg5), autophagy-related gene (Atg) 6 (Beclin-1), apoptosis protein B-cell lymphoma 2 (Bcl-2), Bcl-2-accociated protein (Bax), cysteinyl aspartate specific proteinase (cleaved-caspase-3) proteins expression. Results Compared with the C group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the mRNA relative expressions and protein phosphorylation levels of JAK2, STAT3 in the Infect group were obviously decreased (P<0.05); the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-caspase-3 were obviously increased (P<0.05). Compared with Infect group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the mRNA relative expressions and protein phosphorylation levels of JAK2, STAT3 in the Infect+L-NGR1 group and Infect+H-NGR1 group were obviously increased (P<0.05); the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-Caspase-3 were obviously decreased (P<0.05). Compared with Infect+H-NGR1 group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the protein phosphorylation levels of JAK2, STAT3 in the Infect+H-NGR1+SD-1029 group were obviously decreased (P<0.05), and the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-caspase-3 were obviously increased (P<0.05). Conclusions NGR1 can activate the JAK2/STAT3 signaling pathway, promote autophagy of alveolar epithelial cells, and inhibit Kp-induced inflammatory injury and apoptosis of alveolar epithelial cells.

8.
Article in Chinese | WPRIM | ID: wpr-1005808

ABSTRACT

【Objective】 To investigate the influence of matrine (MT) on the balance of T helper cell 17 (Th17)/regulatory T cells (Treg) in rats with inflammatory bowel disease by regulating interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (STAT3)/nuclear transcription factor-κB (NF-κB) pathway. 【Methods】 SD rats were grouped into control check group (CK group), model group, low-dose MT group (MT-L group, 50 mg/kg), medium-dose MT group (MT-M group, 100 mg/kg), high-dose MT group (MT-H group, 200 mg/kg), mesalazine group (MSLM group, 0.42 g/kg), and MT-H+rIL-6 (IL-6 activator) group (200 mg/kg+0.05 mg/kg) according to the random number table method, with 18 in each group. Except for the CK group, the rats in other groups all received with 5% trinitrobenzenesulfonic acid (20 mg/kg) buffer solution mixed with 50% ethanol at a ratio of 1∶1 and then enema to construct a rat model of inflammatory bowel disease. After the successful modeling, they were treated with drug administration once a day for 7 weeks. The body weight of rats was measured at 1, 3, 5, and 7 weeks of administration; the changes of colon length of rats in each group were compared; HE staining was used to detect the pathological damage of rat colon tissue; the levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-17 and IL-10 in serum of rats were detected by ELISA; the proportions of Th17 and Treg cells in peripheral blood of rats were detected by flow cytometry; Western blottingt was performed to detect the protein expression of retinoic acid-related orphan receptor γt (RORγt), forkhead box protein P3 (Foxp3), IL-6, p-STAT3, and p-NF-κB p65 in rat colon tissue. 【Results】 Compared with the CK group, the colon tissue of the model group was severely damaged by pathology, and the body weight (at 3, 5, and 7 weeks), the level of IL-10, the proportion of Treg cell, and the expression of Foxp3 protein were decreased, the colon length shortened, the levels of TNF-α, IL-17, the proportions of Th17 cell, Th17/Treg ratio, and the protein expression of RORγt, IL-6, p-STAT3, and p-NF-κB p65 increased (P<0.05). Compared with the model group, the corresponding indicators of the MT-L group, MT-M group, MT-H group, and MSLM group had the opposite trends (P<0.05); rIL-6 attenuated the promoting effect of high-dose MT on Th17/Treg balance in inflammatory bowel disease rats. 【Conclusion】 MT may promote Th17/Treg balance in inflammatory bowel disease rats by inhibiting IL-6/STAT3/NF-κB signaling pathway.

9.
Article in Chinese | WPRIM | ID: wpr-1005770

ABSTRACT

【Objective】 To investigate the role of LIF/LIFR/STAT3 pathway in endometrial receptivity in rats with polycystic ovary syndrome (PCOS). 【Methods】 Forty 21-day-old SD female rats were divided into normal (control) group, model group, sham-operation group, and LIF group with 10 rats in each. The rat model of PCOS was constructed by subcutaneous injection of prasterone sodium sulfate at the back of the neck. The serum levels of testosterone (T), glucose and insulin in each group were detected. The morphological changes of the uterus in each group were observed by HE staining, and the morphological changes of endometrium were measured. Immunohistochemistry, Western blotting, and Real-time fluorescence quantitative PCR(qRT-PCR) were used to determine the protein expression and mRNA expression of LIF and STAT3 in rat endometrium. 【Results】 Compared with control group, the levels of integrin avb3, serum T, insulin and glucose in PCOS rats were significantly increased (P=0.000, P=0.000, P=0.001). Supplementation of exogenous LIF could significantly reduce the levels of integrin avb3, serum T, glucose and insulin in PCOS rats (P=0.000, P=0.002, P=0.003, P=0.007). HE results showed that exogenous LIF could reduce uterine cavity and glandular morphology in PCOS rats and increase the equivalent diameter (P=0.000, P=0.000) and area (P=0.000, P=0.000) of uterine glands and glandular cavity, the ratio of glandular interstitial area (P=0.000), and the average endometrial thickness (P=0.006), with statistically significant differences. Immunohistochemistry, Western blotting, and qRT-PCR results showed that the expression levels of LIF and p-STAT3 protein and mRNA in model group were significantly decreased compared with control group. Compared with model group, the protein and mRNA expressions of LIF and p-STAT3 in LIF group were significantly increased (P<0.05). 【Conclusion】 Exogenous LIF supplementation can improve endometrial receptivity in PCOS rats, and its mechanism is related to the LIF/LIFR/STAT3 pathway.

10.
Article in Chinese | WPRIM | ID: wpr-998748

ABSTRACT

Background Aluminum activates signal transducer and activator of transcription 3 (STAT3), causing microglial nucleotide-binding and oligomerization domain-like receptors protein 3 (NLRP3) inflammasome activation and inflammatory responses and producing neurotoxicity. Objective To explore the role of STAT3 regulated NLRP3 inflammasomes in the inflammatory response of mouse microglia cell line (BV2) cells induced by maltol aluminum [Al(mal)3]. Methods BV2 cells were assigned to five groups: one control group, three Al(mal)3 exposure groups (low, medium, and high doses at 40, 80, and 160 μmol·L−1 Al(mal)3 respectively), and one C188-9 (STAT3 antagonist) intervention group [10 μmol·L−1 C188-9 +160 μmol·L−1 Al(mal)3]. Cell viability was detected by CCK8. The expression of M1/M2 type markers, i.e. CD68/CD206, STAT3, p-STAT3, NLRP3, cleaved-casepase-1, and apoptosis-associated speck-like protein (ASC) in BV2 cells were detected by Western blotting, and proinflammatory cytokines interleukin (IL)-1β and IL-18, and anti-inflammatory cytokine IL-10 were determined by ELISA. Results The results of cell viability assay showed that cell viability gradually decreased with the increase of Al(mal)3 dose. Compared with the control group, the cell viability of the Al(mal)3 high-dose group was decreased by 18% (P<0.05); compared with the Al(mal)3 high-dose group, the cell viability of the C188-9 intervention group was significantly elevated by 14% (P<0.05). Compared with the control group, the expression levels of CD68 in the Al(mal)3 low-, medium-, and high-dose groups were elevated by 19%, 20%, and 21%, respectively (P<0.05); the expression level of CD206 in the Al(mal)3 high-dose group was decreased by 25% (P<0.05). Compared with the Al(mal)3 high-dose group, the expression level of CD68 in the C188-9 intervention group was reduced by 9% (P<0.05), whereas the expression level of CD206 was elevated by 22% (P<0.05). Compared with the control group, the p-STAT3 protein expression and the p-STAT3/STAT3 ratio in the Al(mal)3 high-dose group increased by 129% and 127%, respectively (P<0.05). Compared with the Al(mal)3 high-dose group, the p-STAT3 protein expression and the p-STAT3/STAT3 ratio in the C188-9 intervention group were decreased by 55% and 54%, respectively (P>0.05). Compared with the control group, the expression level of NLRP3 protein increased by 75% in the Al(mal)3 high-dose group (P<0.05), the expression levels of cleaved-casepase-1 protein increased by 28% and 35% in the Al(mal)3 medium- and high-dose groups (P<0.05), and the expression levels of ASC increased by 22%, 25%, and 53% in the Al(mal)3 low-, medium- and high-dose groups (P<0.05), respectively. Compared with the Al(mal)3 high-dose group, the expression levels of NLRP3, cleaved-casepase-1, and ASC proteins in the C188-9 intervention group decreased by 30%, 19%, and 32%, respectively (P<0.05). Compared with the control group, the levels of IL-1β in the Al(mal)3 medium- and high-dose groups increased by 18% and 21%, respectively (P<0.05), and the level of IL-18 in the Al(mal)3 high-dose group increased by 10% (P<0.05). Compared with the Al(mal)3 high-dose group, the IL-18 levels were reduced by 23% in the C188-9 intervention group (P<0.05). The content of anti-inflammatory factor IL-10 did not differ significantly between groups (P>0.05). Conclusion Aluminum can induce inflammatory responses in BV2 microglia and is predominantly pro-inflammatory, and the mechanism may involve STAT3 regulation of NLRP3 inflammasome secretion of inflammatory factors.

11.
Article in Chinese | WPRIM | ID: wpr-969607

ABSTRACT

ObjectiveTo study the correlation between Vimentin and hepatocellular carcinoma (HCC) and the mechanism of Jianpi Yiqi prescription against HCC through Vimentin. MethodCorrelation between Vimentin and HCC was analyzed based on the cancer genome atlas(TCGA), clinical proteomic tumor analysis consortium(CPTAC), STRING, and Cytoscape. SD rats were randomized into normal group (normal saline, ig, once/day, 4 weeks), model group (normal saline, ig, once/day, 4 weeks), low-dose, medium-dose, and high-dose (5.25, 10.5, 21 g·kg-1, ig, once/day, 4 weeks) JianpiYiqi prescription groups, signal transducer and activator of transcription 3 (STAT3) inhibition group (C188-9, 4.5 mg·kg-1, ip, once/day, 4 weeks), and glycoprotein 130 (gp130) inhibition group (SC144, 4.5 mg·kg-1, ip, once/day, 4 weeks), 10 rats in each group. Diethylnitrosamine (DEN, 70 mg·kg-1 body weight, ip) was injected in rats except the normal group to induce HCC. After the modeling, administration began. After last administration, Real-time polymerase chain reaction(Real-time PCR) was performed to determine Vimentin mRNA level in rat liver tissue. Caspase-3 activity in liver tissue was detected by colorimetry, and expression of Rho kinase (ROCK)1, ROCK2, aurora kinase B (AURKB), Zinc-finger protein 148 (ZNF148)/zinc-binding protein-89 (ZBP-89), STAT3, p-STAT3, total Vimentin, and phosphorylated (p)-Vimentin in liver tissue and Vimentin in liver tissue nucleus detected by Western blot. Serum Vimentin concentration was measured by enzyme-linked immunosorbent assay (ELISA). ResultVimentin mRNA level was high in tissues from HCC patients with different cancer stages (stage Ⅰ-Ⅳ), different pathological grades (G1-G3), no regional lymph node metastasis (N0), and different subtypes (P<0.01). Vimentin mRNA expression was higher in tissues from patients with lymph node metastasis than in patients without lymph node metastasis and normal samples. Vimentin protein level was decreased in HCC tissues (P<0.01). Vimentin gene has 4 mutations which can induce change in the primary structure of Vimentin protein and patients with Vimentin gene mutation had short disease free survival time (P<0.01). The mRNA expression of Vimentin was negatively associated with HCC cell purity (P<0.01) but was positively associated with the infiltration levels of cancer-associated fibroblasts, M2 macrophages, myeloid dendritic cell and other immune cells in tumor microenvironment (P<0.01). Association analysis results showed that the expression of Vimentin was correlated with the STAT3 expression in HCC tissues (P<0.01). As for the animal experiment, Vimentin mRNA level and protein levels of total Vimentin and p-Vimentin in liver tssue, Vimentin protein level in liver tissue nucleus, Vimentin in rat serum, ROCK2, AURKB, STAT3 and p-STAT3 in liver tissues were up-regulated (P<0.01) and protein level of negative regulator ZBP-89 was reduced in the model group (P<0.01) compared with those in the normal group. Activity of Caspase-3 in liver tissue increased and the ROCK1 protein level was increased in the model group compared with those in the normal group. STAT3 inhibitor, gp130 inhibitor, and medium-dose and high-dose Jianpi Yiqi prescription all can reduce the secretory Vimentin protein in serum, protein levels of total Vimentin and p-Vimentin in liver tissues, and Vimentin in liver tissue nucleus, and the protein levels of STAT3/Vimentin signaling pathway-related molecules, such as STAT3, p-STAT3, ROCK2, and AURKB and up-regulate the protein level of negative regulator ZBP-89 and activity of Caspase-3 (P<0.05, P<0.01). Effect of medium-dose or high-dose Jianpi Yiqi prescription on Vimentin mRNA expression, STAT3 protein expression, ZBP-89 protein expression, ROCK2 protein expression, AURKB protein expression and Caspase-3 activity was not significantly different from that of STAT3 inhibitor. ConclusionVimentin, an important inflammatory molecule, is closely related to the occurrence and development of HCC and its expression, subcellular location and function may be affected by cancer-associated fibroblasts, M2 macrophages, myeloid dendritic cell, and IL-6/STAT3 signaling pathway, particularly by STAT3 molecule. Jianpi Yiqi prescription may exert therapeutic effect on HCC via regulating Vimentin through the STAT3/Vimentin signaling pathway.

12.
China Pharmacy ; (12): 819-824, 2023.
Article in Chinese | WPRIM | ID: wpr-969578

ABSTRACT

OBJECTIVE To study the improvement effects and mechanism of rhein on immunoglobulin A nephropathy (IgAN) model rat based on signal transducer and activator of transcription 3 (STAT3) signaling pathway. METHODS Rats were randomly divided into normal control group, IgAN model group and rhein treatment group, with 10 rats in each group. IgAN model group and rhein treatment group were given combination of bovine serum albumin+lipopolysaccharide+carbon tetrachloride to induce IgAN model. Since the 7th week, rhein treatment group rats were intragastrically given relevant medicine, and normal control group and model group rats were given equal amount of normal saline intragastrically, for consecutive 4 weeks. After the last administration, the count of urine sediment erythrocyte, 24 h-urine total protein (UTP), the levels of immunoglobulin A (IgA) in serum and secretory immunoglobulin A (sIgA) in intestinal mucosa were detected. The pathological changes of Peyer’s patch in renal cortex and intestinal mucosa and IgA deposition in renal cortex were observed. The expressions of interleukin-17 (IL-17), IL- 6 and transforming growth factor β (TGF-β) in Peyer’s patch of intestinal mucosa in rats were detected. The expressions of STAT3 and related orphan receptor γt (RORγt) mRNA in Peyer’s patch were detected. The expressions of p-STAT3 and RORγt proteins in Peyer’s patch were detected. RESULTS Compared with normal control group, the count of urine sediment erythrocyte, 24 h-UTP, the levels of IgA in serum and sIgA in intestinal mucosa were increased significantly in IgAN model group (P<0.01); enlarged renal corpuscles, dilated renal sacs, obvious intratubular mesangial hyperplasia and fibrosis were observed in renal cortex; the volume and germinal center of Peyer’s patch in intestinal mucosa increased; IgA deposition of renal cortex zxyylxk20220103) was obvious; the expressions of IL-17, IL-6 and TGF-β in Peyer’s patch, mRNA expressions of STAT3 and RORγt, protein expressions of p-STAT3 and RORγt were increased significantly (P<0.01). Compared with IgAN model group,above indexes were decreased significantly in rhein treatment group (P<0.01), pathological damage of renal cortex was improved, the volume of Peyer’s patch and germinal center of intestinal mucosa were reduced, and IgA deposition in renal cortex was weakened. CONCLUSIONS Rhein can improve IgAN model rats, the mechanism of which may be associated with inhibiting STAT3 signaling pathway and regulating immune function of Peyer’s patch in intestinal mucosa.

13.
Article in Chinese | WPRIM | ID: wpr-965647

ABSTRACT

ObjectiveTo explore the role of interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (STAT3) pathway in the balance of T helper 17 (Th17)/regulatory T (Treg) cells in ulcerative colitis (UC) with internal dampness-heat accumulation syndrome and the intervention mechanism of Shaoyaotang. MethodA total of 60 SD rats were randomized into blank group (equivalent volume of normal saline), model group (equivalent volume of normal saline), western medicine control group (0.42 g·kg-1 mesalazine), and low-dose (11.1 g·kg-1), medium-dose (22.2 g·kg-1), and high-dose (44.4 g·kg-1) Shaoyaotang groups. UC with internal dampness-heat accumulation syndrome was induced in rats with the compound method except for the blank group. The administration lasted 14 days for each group. At 24 h after the last administration, rats were killed and the spleen and colon tissues were separated. The histopathological changes of colon were observed based on hematoxylin and eosin (HE) staining and the levels of interleukin-17 (IL-17) and transforming growth factor-β1 (TGF-β1) in colon tissue were detected by immunohistochemistry (IHC). Flow cytometry was employed to determine the levels of Th17/Treg cells in the spleen, and Western blot to measure the levels of IL-6 and STAT3 proteins in colon tissue. ResultCompared with the blank group, the model group had lesions such as congestion and erosion, low percentage of spleen Treg cells (P<0.01), high percentage of Th17 cells (P<0.01), and high levels of IL-6 and STAT3 proteins in colon tissue (P<0.01). Compared with the model group, the administration groups showed alleviation of colon injury, high percentage of spleen Treg cells (P<0.05, P<0.01), low percentage of Th17 cells (P<0.01), and low levels of IL-6 and STAT3 proteins in colon tissue (P<0.01). ConclusionShaoyaotang regulates the balance of Th17/Treg by inhibiting the IL-6/STAT3 pathway, thereby relieving the pathological damage of UC rats with internal dampness-heat accumulation syndrome and affecting their immune function.

14.
Article in Chinese | WPRIM | ID: wpr-976534

ABSTRACT

To investigate the therapeutic effect and mechanism of Qingwen Baiduyin on acute lung injury (ALI) in mice induced by lipopolysaccharide (LPS). MethodA total of 144 C57BL/6 mice were randomly divided into the following groups: a normal group, a model group (LPS, 5 mg·kg-1), a dexamethasone group (5 mg·kg-1), and low-, medium-, and high-dose Qingwen Baiduyin groups (14.105, 28.21, 56.42 g·kg-1). The mice were treated once daily for 5 days. One hour after the final administration, the ALI model was established by intratracheal instillation of LPS, and samples were collected at 6 h and 24 h after modeling. The arterial blood gas index of mice was analyzed. The total protein content, total cell count, Evans blue dye (EBD) content, and lung tissue wet-to-dry weight ratio (W/D) of bronchoalveolar lavage fluid (BALF) were measured. Hematoxylin-eosin (HE) staining was performed to assess the pathological changes in mouse lung tissue. Western blot was used to detect the expression levels of key proteins in the Janus kinase 1/signal transducer and activator of transcription 1/interferon regulatory factor 1 (JAK1/STAT1/IRF1) signaling pathway in lung tissue. ResultCompared with the normal group, the model group showed reduced arterial oxygen pressure (pO2), oxygen saturation (SO2), and lung tissue W/D (P<0.05, P<0.01), increased carbon dioxide pressure (pCO2), total protein content, total cell count, EBD content, interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), chemokine CXC ligand 1 (CXCL1), chemokine CXC ligand 2 (CXCL2), chemokine CXC ligand 9 (CXCL9), and chemokine CXC ligand 10 (CXCL10) content (P<0.05, P<0.01), thickening of the alveolar walls, fusion of alveolar cavities, and infiltration of inflammatory cells in lung tissue, increased proportion of M1 macrophage polarization and lung cell apoptosis (P<0.05), and increased protein expression levels of JAK1, phosphorylated JAK1 (p-JAK1), inducible nitric oxide synthase (iNOS), STAT1, phosphorylated STAT1 (p-STAT1), IRF1, gasdermin D (GSDMD), and mixed lineage kinase domain-like protein (MLKL) (P<0.05, P<0.01). Compared with the model group, Qingwen Baiduyin significantly increased pO2, SO2, and lung tissue W/D (P<0.05, P<0.01), improved the pathological changes in lung tissue, and reduced pCO2, total protein content, total cell count, EBD content, IFN-γ, TNF-α, IL-1β, CXCL1, CXCL2, CXCL9, and CXCL10 content, proportion of M1 macrophage polarization, and protein expression levels of JAK1, p-JAK1, iNOS, STAT1, p-STAT1, IRF1, GSDMD, and MLKL (P<0.05, P<0.01). ConclusionQingwen Baiduyin can improve the lung inflammatory response and reduce lung cell apoptosis in mice with ALI by inhibiting the JAK1/STAT1/IRF1 signaling pathway, thereby exerting a lung-protective effect.

15.
Article in Chinese | WPRIM | ID: wpr-973746

ABSTRACT

ObjectiveTo observe the effect of Hedysari Radix polysaccharide (HRP) on the Janus kinase 2 (JAK2)/signal transducer and activator of transcription protein 3 (STAT3) signaling pathway in diabetic nephropathy db/db mice. MethodFifty db/db mice were randomly divided into model group, irbesartan group (irbesartan suspension, 22.75 mg·kg-1), and high-, medium-, and low-dose HRP groups (HRP suspension, 200, 100, 50 mg·kg-1) according to the body weight, with 10 mice in each group. Another 10 C57BL/6 mice were assigned to the normal group. The mice were treated with corresponding drugs by gavage, while those in the normal group and the model group received distilled water at 5 mL·kg-1. The mice in the six groups were administered once a day by gavage for 12 consecutive weeks. The uric acid (UA), triglycerides (TG), and total cholesterol (TC) were detected. Periodic acid-Schiff (PAS) staining and Masson staining were used to observe the pathological changes in kidney tissues. Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect the protein and mRNA expression levels of JAK2, STAT3, suppressor of cytokine signaling 3 (SOCS3), and tumor necrosis factor-α (TNF-α) in the kidney. ResultAfter 12 weeks of treatment, compared with the normal group, the model group showed significant pathological ultrastructural changes in kidney tissues and increased UA, TG, and TC levels (P<0.01). Compared with the model group, the high- and medium-dose HRP groups and the irbesartan group showed improvement in pathological ultrastructure of kidney tissues and reduced UA, TG, and TC levels (P<0.05, P<0.01). Compared with the normal group, the model group showed a decrease in SOCS3 protein and mRNA expression levels and an increase in JAK2, STAT3, and TNF-α protein and mRNA expression levels (P<0.01). Compared with the model group, the high- and medium-dose HRP groups and the irbesartan group showed an increase in SOCS3 protein and mRNA expression levels and a decrease in JAK2, STAT3, and TNF-α protein and mRNA expression levels (P<0.05, P<0.01). ConclusionHRP can alleviate renal damage in diabetic nephropathy to a certain extent, and its mechanism may be related to the inhibition of the activation of the JAK2/STAT3 signaling pathway.

16.
Article in Chinese | WPRIM | ID: wpr-973740

ABSTRACT

ObjectiveTo investigate the effect of Qiling Baitouweng Tang (QLBTWT) on proliferation and apoptosis, Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway and interleukin-10 (IL-10) in diffuse large B-cell lymphoma (DLBCL). MethodWith human DLBCL cells OCI-LY10 and U2932 as research objects, cell proliferation was detected by cell counting kit-8 (CCK-8) assay. After treatment with 0, 4.6, 9.3, 18.7, 37.5, 75, 150 mg·L-1 QLBTWT for 24 h, the half-inhibitory concentration (IC50) of OCL-LY10 and U2932 cells was calculated to be 9.33, 16.13 mg·L-1, respectively, based on which, 9.5, 19, 38 mg·L-1 QLBTWT were selected for subsequent experiments. After 0, 9.5, 19, 38 mg·L-1 QLBTWT treatment for 24 h, the zymogen activities of Caspase-3, Caspase-8 and Caspase-9 in OCI-LY10 and U2932 cells were detected using corresponding activity assay kits (colorimetric), and the IL-10 expression was detected by enzyme-linked immuno sorbent assay (ELISA). The apoptosis rate and cell cycle of OCI-LY10 and U2932 cells treated with different concentrations of QLBTWT for 24 h were detected by flow cytometry. The expressions of apoptosis-related proteins [B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), cleaved poly adenosine diphosphate ribose polymerase (cleaved PARP), cleaved Caspase-3], JAK2, STAT3, phospho-JAK2 (p-JAK2), phospho-STAT3 (p-STAT3) pathway proteins, and c-Myc protein in OCL-LY10 and U2932 cells after 24 h treatment with 0, 9.5, 19, 38 mg·L-1 QLBTWT were all tested by Western blot. ResultAfter QLBTWT treatment on OCI-LY10 and U2932 cells for 24 h, cell proliferation was inhibited in each QLBTWT group compared with that in the control group (P<0.05, P<0.01). The zymogens of Caspase-3, Caspase-8 and Caspase-9 were activated (P<0.01), and there was an increase in cell apoptosis (P<0.05, P<0.01) and cell cycle arrest at Gap phase1 (G1) phase in 9.5, 19 and 38 mg·L-1 QLBTWT group (P<0.05, P<0.01). After 9.5, 19 and 38 mg·L-1 QLBTWT treatment on OCI-LY10 and U2932 cells for 24 h, the expressions of Bcl-2, p-JAK2 and p-STAT3 proteins were decreased (P<0.01), and the expressions of Bax, cleaved PARP and cleaved Caspase-3 proteins were increased (P<0.01), but no significant change was observed in the expressions of JAK2 and STAT3 proteins. Compared with the conditions in the control group, the expressions of c-Myc, p-JAK2, and p-STAT3 proteins were down-regulated in 19 mg·L-1 QLBTWT group and 19 mg·L-1 QLBTWT+10 μg·L-1 IL-10 group (P<0.05, P<0.01), and up-regulated in 10 μg·L-1 IL-10 group (P<0.05, P<0.01), while there was no difference in JAK2/STAT3 proteins. ConclusionQLBTWT can inhibit proliferation and induce apoptosis of human DLBCL cells OCI-LY10 and U2932, and the potential mechanism may be related to the regulation of JAK2/STAT3 signaling pathway.

17.
Article in Chinese | WPRIM | ID: wpr-1015605

ABSTRACT

Interleukin-6 (IL-6) is a spreading pleiotropic cytokine, with both anti-inflammatory and proinflammatory effects. It not only participates in the body immune responses but also is involved in the biological regulative processes among different organs, tissues, and cells. IL-6 has both anti-inflammatory and pro-inflammatory effects. In the early stage of pathogen infection, IL-6 plays an anti-inflammatory role in the body, and its level is moderately increased in the body to resist inflammation and maintain internal homeostasis. However, a large amount of IL-6 release can cause excessive inflammation and trigger other pathological changes in the body. Il-6 also has the dual effect of stimulating the synthesis and degradation of skeletal muscle protein in regulating skeletal muscle mass. As an important locomotive organ, skeletal muscle is also one of the key targets of IL-6. IL-6 takes part in the biological control of skeletal muscle hypertrophy through regulating muscle satellite cell proliferation and differentiation under specific stresses. In addition IL-6 is also associated with skeletal muscle atrophy induced by aging and other pathological stresses. In addition, during exercise stress, skeletal muscle can also serve as an endocrine organ to secrete and release IL-6 that facilitates the "crosstalk" between skeletal muscle and other organs or tissues. As IL-6 plays as a versatile role in our body, this paper reviews the research progress of the mechanism of IL-6 in the regulation of skeletal muscle mass, which may provide theoretical support for revealing the molecular mechanism of skeletal muscle stresses and adaptations.

18.
Article in English | WPRIM | ID: wpr-1010315

ABSTRACT

OBJECTIVE@#To explore the anti-inflammatory effects of ethyl lithospermate in lipopolysaccharide (LPS)-stimulated RAW 264.7 murine-derived macrophages and zebrafish, and its underlying mechanisms.@*METHODS@#3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazoliumbromide (MTT) assays were performed to investigate the toxicity of ethyl lithospermate at different concentrations (12.5-100 µ mol/L) in RAW 264.7 cells. The cells were stimulated with LPS (100 ng/mL) for 12 h to establish an inflammation model in vitro, the production of pro-inflammatory cytokines interleukin (IL)-6 and tumor necrosis factor α (TNF-α) were assessed by enzyme linked immunosorbent assay (ELISA). Western blot was used to ascertain the protein expressions of signal transducer and activator of transcription 3 (STAT3), nuclear factor kappa B (NF-κB) p65, phospho-STAT3 (p-STAT3, Tyr705), inhibitor of NF-κB (IκB) α, and phospho-I κB α (p-IκB α, Ser32), and confocal imaging was used to identify the nuclear translocation of NF-κB p65 and p-STAT3 (Tyr705). Additionally, the yolk sacs of zebrafish (3 days post fertilization) were injected with 2 nL LPS (0.5 mg/mL) to induce an inflammation model in vivo. Survival analysis, hematoxylin-eosin (HE) staining, observation of neutrophil migration, and quantitative real-time polymerase chain reaction (qRT-PCR) were used to further study the anti-inflammatory effects of ethyl lithospermate and its probable mechanisms in vivo.@*RESULTS@#The non-toxic concentrations of ethyl lithospermate have been found to range from 12.5 to 100 µ mol/L. Ethyl lithospermate inhibited the release of IL-6 and TNF-α(P<0.05 or P<0.01), decreased IκBα degradation and phosphorylation (P<0.05) as well as the nuclear translocation of NF-κB p65 and p-STAT3 (Tyr705) in LPS-induced RAW 264.7 cells (P<0.01). Ethyl lithospermate also decreased inflammatory cells infiltration and neutrophil migration while increasing the survival rate of LPS-stimulated zebrafish (P<0.05 or P<0.01). In addition, ethyl lithospermate also inhibited the mRNA expression levels of of IL-6, TNF-α, IκBα, STAT3, and NF-κB in LPS-stimulated zebrafish (P<0.01).@*CONCLUSION@#Ethyl lithospermate exerts anti-Inflammatory effected by inhibiting the NF-κB and STAT3 signal pathways in RAW 264.7 macrophages and zebrafish.


Subject(s)
Animals , Mice , NF-kappa B/metabolism , Lipopolysaccharides , Zebrafish , NF-KappaB Inhibitor alpha/metabolism , Interleukin-6/metabolism , Tumor Necrosis Factor-alpha/metabolism , STAT3 Transcription Factor/metabolism , Inflammation/metabolism , Anti-Inflammatory Agents/therapeutic use
19.
Article in Chinese | WPRIM | ID: wpr-982061

ABSTRACT

OBJECTIVE@#To investigate the effect of kaempferol on proliferation of acute myeloid leukemia (AML) KG1a cells and its mechanism.@*METHODS@#Human AML KG1a cells in logarithmic growth stage were taken and set at 25, 50, 75 and 100 μg/ml kaempferol group, another normal control group (complete medium without drug) and solvent control group (add dimethyl sulfoxide) were also set. After 24 and 48 hours of intervention, the cell proliferation rate was detected by CCK-8 assay. In addition, interleukin-6 (IL-6) combined with kaempferol group (Plus 20 μg/l IL-6 and 75 μg/ml kaempferol) was set up, 48 hours after culture, the cell cycle and apoptosis of KG1a cells were detected by flow cytometry, the mitochondrial membrane potential (MMP) of KG1a cells was detected by MMP detection kit (JC-1 method), and the expression of Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) pathway related proteins in KG1a cells were detected by Western blot.@*RESULTS@#The cell proliferation rate of 25, 50, 75 and 100 μg/ml kaempferol group decreased significantly (P<0.05), and with the increase of kaempferol dose (r24 h=-0.990, r48 h= -0.999), the cell proliferation rate decreased gradually (P<0.05). The inhibitory effect of 75 μg/ml kaempferol on cell proliferation reached half of effective dose after 48 hours of intervention. Compared with normal control group, the G0/G1 phase cell proportion and apoptosis rate of cells in 25, 50 and 75 μg/ml kaempferol group increased, while the S phase cell proportion, MMP, phosphorylated JAK2 (p-JAK2)/JAK2 and phosphorylated STAT3 (p-STAT3)/STAT3 protein expression decreased in a dose-dependent manner (r=0.998, 0.994, -0.996, -0.981, -0.997, -0.930). Compared with 75 μg/ml kaempferol group, the G0/G1 phase cell proportion and apoptosis rate of cells in IL-6 combined with kaempferol group decreased, while the S phase cell proportion, MMP, p-JAK2/JAK2 and p-STAT3/STAT3 protein expression increased significantly (P<0.05).@*CONCLUSION@#Kaempferol can inhibit KG1a cell proliferation and induce KG1a cell apoptosis, its mechanism may be related to the inhibition of JAK2/STAT3 signal pathway.


Subject(s)
Humans , STAT3 Transcription Factor/metabolism , Interleukin-6/metabolism , Kaempferols/pharmacology , Signal Transduction , Apoptosis , Janus Kinase 2 , Cell Proliferation , Leukemia, Myeloid, Acute
20.
Article in Chinese | WPRIM | ID: wpr-981456

ABSTRACT

Based on the Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3) signaling pathway, this study investigated the effect of medicated serum of Sparganii Rhizoma(SR) and Curcumae Rhizoma(CR) on the proliferation, apoptosis, migration, and secretion of inflammatory factors of ectopic endometrial stromal cells(ESCs). Specifically, human ESCs were primary-cultured. The effect of different concentration(5%, 10%, 20%) of SR-, CR-, and SR-CR combination-medicated serum, and AG490 solution(50 μmol·L~(-1)) on the proliferation of ESCs was detected by methyl thiazolyl tetrazolium(MTT) assay, and the optimal dose was selected accordingly for further experiment. The cells were classified into normal serum(NS) group, SR group(10%), CR group(10%), combination(CM) group(10%), and AG490 group. The apoptosis level of ESCs was detected by flow cytometry, and the migration ability was examined by wound healing assay. The secretion of interleukin(IL)-1β, IL-6, and tumor necrosis factor(TNF)-α was determined by enzyme-linked immunosorbent assay(ELISA). The protein levels of cysteinyl aspartate specific protei-nase-3(caspase-3), B-cell lymphoma(Bcl-2), and Bcl-2-associated X protein(Bax) and the levels of phosphorylated(p)-JAK2 and p-STAT3 were detected by Western blot. The results showed that the viability of ESCs cells was lowered in the administration groups compared with the blank serum group(P<0.01), especially the 10% drug-medicated serum, which was selected for further experiment. The 10% SR-medicated serum, 10% CR-medicated serum, and 10% CM-medicated serum could increase the apoptosis rate(P<0.01), up-regulate the protein expression of caspase-3 and Bax in cells(P<0.05 or P<0.01), down-regulate the expression of Bcl-2(P<0.01), decrease the cell migration rate(P<0.05 or P<0.01), and reduce the secretion levels of IL-1β, IL-6, and TNF-α(P<0.05 or P<0.01), and levels of p-JAK2 and p-STAT3(P<0.05 or P<0.01). Compared with the SR and CR groups, CM group showed low cell viability(P<0.01), high protein expression of caspase-3 and Bax(P<0.05 or P<0.01), and low protein expression of Bcl-2 and p-JAK2(P<0.05). After incubation with CM, the apoptosis rate was higher(P<0.05) and the migration rate was lower(P<0.01) than that of the CR group. The p-STAT3 protein level of CM group was lower than that of the RS group(P<0.05). The mechanism of SR, CR, and the combination underlying the improvement of endometriosis may be that they blocked JAK2/STAT3 signaling pathway, inhibited ESC proliferation, promoted apoptosis, weakened cell migration, and reduced the secretion of inflammatory factors. The effect of the combination was better than that of RS alone and CR alone.


Subject(s)
Female , Humans , Janus Kinase 2 , Caspase 3 , bcl-2-Associated X Protein , Interleukin-6/genetics , Apoptosis , Signal Transduction , Cell Proliferation , STAT3 Transcription Factor/genetics
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