Lycium barbarum polysaccharide's protective effects against PM2.5-induced cellular senescence in HUVECs.

Fine particulate matter (PM2.5) exposure is strongly associated with vascular endothelial senescence, a process implicated in cardiovascular diseases. While there is existing knowledge on the impact of Lycium barbarum polysaccharide (LBP) on vascular endothelial damage, the protective mechanism of LBP against PM2.5-induced vascular endothelial senescence remains unclear. In this study, we investigated the impact of PM2.5 exposure on vascular endothelial senescence and explored the intervention effects of LBP in human umbilical vein endothelial cells (HUVECs). We found that PM2.5 exposure dose-dependently reduced cell viability and proliferation in HUVECs while increasing the production of reactive oxygen species (ROS), malondialdehyde (MDA), and hydrogen peroxide (H2O2). Additionally, PM2.5 exposure inhibited the activity of superoxide dismutase (SOD). Notably, PM2.5 exposure induced autophagy impairments and cellular senescence. However, LBP mitigated PM2.5-induced cell damage. Further studies demonstrated that correcting autophagy impairment in HUVECs reduced the expression of the senescence markers P16 and P21 induced by PM2.5. This suggests the regulatory role of autophagy in cellular senescence and the potential of LBP in improving HUVECs senescence. These findings provide novel insights into the mechanisms underlying PM2.5-induced cardiovascular toxicity and highlight the potential of LBP as a therapeutic agent for improving vascular endothelial health.

Effects of gene polymorphisms on delayed MTX clearance, toxicity, and metabolomic changes after HD-MTX treatment in children with acute lymphoblastic leukemia.

This study aims to assess the role of methotrexate-related gene polymorphisms in children with acute lymphoblastic leukemia (ALL) during high-dose methotrexate (HD-MTX) therapy and to explore their effects on serum metabolites before and after HD-MTX treatment. The MTHFR 677C>T, MTHFR 1298A>C, ABCB1 3435C>T, and GSTP1 313A>G genotypes of 189 children with ALL who received chemotherapy with the CCCG-ALL-2020 regimen from January 2020 to April 2023 were analyzed, and toxic effects were reported according to the Common Terminology Criteria for Adverse Events (CTCAE, version 5.0). Fasting peripheral blood serum samples were collected from 27 children before and after HD-MTX treatment, and plasma metabolites were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS). The results of univariate and multivariate analyses showed that MTHFR 677C>T and ABCB1 3435 C>T gene polymorphisms were associated with the delayed MTX clearance (P < 0.05) and lower platelet count after treatment in children with MTHFR 677 mutation compared with wild-type ones (P < 0.05), and pure mutations in ABCB1 3435 were associated with higher serum creatinine levels (P < 0.05). No significant association was identified between MTHFR 677C>T, MTHFR 1298A>C, ABCB1 3435 C>T, and GSTP1 313A>G genes and hepatotoxicity or nephrotoxicity (P > 0.05). However, the serum metabolomic analysis indicated that the presence of the MTHFR 677C > T gene polymorphism could potentially contribute to delayed MTX clearance by influencing L-phenylalanine metabolism, leading to the occurrence of related toxic side effects. CONCLUSION: MTHFR 677C>T and ABCB1 3435 C>T predicted the risk of delayed MTX clearance during HD-MTX treatment in children with ALL. Serum L-phenylalanine levels were significantly elevated after HD-MTX treatment in children with the MTHFR 677C>T mutation gene. TRIAL REGISTRATION: This study was registered at the Chinese Clinical Trial Registry (registration number: ChiCTR2000035264; registration: 2020/08/05; https://www.chictr.org.cn/ ). WHAT IS KNOWN: • MTX-related genes play an important role in MTX pharmacokinetics and toxicity, but results from different studies are inconsistent and the mechanisms involved are not clear. WHAT IS NEW: • Characteristics, prognosis, polymorphisms of MTX-related genes, and metabolite changes were comprehensively evaluated in children treated with HD-MTX chemotherapy. • Analysis revealed that both heterozygous and pure mutations in MTHFR 677C>T resulted in a significantly increased risk of delayed MTX clearance, and that L-phenylalanine has the potential to serve as a predictive marker for the metabolic effects of the MTHFR 677C>T polymorphism.

Hypermethylation of the CTRP9 promoter region promotes Hcy induced VSMC lipid deposition and foam cell formation via negatively regulating ER stress.

To provide a theoretical basis for the prevention and treatment of atherosclerosis (As), the current study aimed to investigate the mechanism underlying the effect of homocysteine (Hcy) on inducing the lipid deposition and foam cell formation of the vascular smooth muscle cell (VSMC) via C1q/Tumor necrosis factor-related protein9 (CTRP9) promoter region Hypermethylation negative regulating endoplasmic reticulum stress (ERs). Therefore, apolipoprotein E deficient (ApoE-/-) mice were randomly divided into the control [ApoE-/- + normal diet (NC)] and high methionine [ApoE-/- + (normal diet supplemented with 1.7% methionine (HMD)] groups (n = 6 mice/group). Following feeding for 15 weeks, the serum levels of Homocysteine (Hcy), total cholesterol (TC), and triglyceride (TG) were measured using an automatic biochemical analyzer. HE and oil red O staining were performed on the aorta roots to observe the pathological changes. Additionally, immunofluorescence staining was performed to detect the protein expression levels of CTRP9, glucose-regulated protein 78 kD (GRP78), phosphorylated protein kinase RNA-like ER kinase (p-PERK), activating transcription factor 6a (ATF6a), phosphorylated inositol-requiring enzyme-1α (p-IRE1α), sterol regulatory element binding proteins-1c (SREBP1c) and sterol regulatory element binding proteins-2 (SREBP2) in VSMC derived from murine aortic roots. In vitro, VSMC was stimulated with 100 µmol/l Hcy. After transfection of plasmids with overexpression and interference of CTRP9, ERs agonist (TM) and inhibitor (4-PBA) were given to stimulate VSMC cells. HE staining and oil red O staining were used to observe the effect of Hcy stimulation on lipid deposition in VSMC. Additionally, The mRNA and protein expression levels of CTRP9, GRP78, PERK, ATF6a, IRE1α, SREBP1c, and SREBP2 in VSMC were detected by RT-qPCR and western blot analysis, respectively. Finally, The methylation modification of the CTRP9 promoter region has been studied. The NCBI database was used to search the promoter region of the CTRP9 gene, and CpG Island was used to predict the methylation site. After Hcy stimulation of VSMC, overexpression of DNMT1, and intervention with 5-Azc, assess the methylation level of the CTRP9 promoter through bisulfite sequencing PCR (BSP). The results showed that the serum levels of Hcy, TC, and TG in the ApoE-/- + HMD group were significantly increased compared with the ApoE-/- + NC group. In addition, HE staining and oil red O staining showed obvious AS plaque formation in the vessel wall, and a large amount of fat deposition in VSMC, thus indicating that the hyperhomocysteinemia As an animal model was successfully established. Furthermore, CTRP9 were downregulated, while GRP78, p-PERK, ATF6a, p-IRE1α, SREBP1c, SREBP2 was upregulated in aortic VSMC in the ApoE-/- + HMD group. Consistent with the in vivo results, Hcy can inhibit the expression of CTRP9 in VSMC and induce ERs and lipid deposition in VSMC. Meanwhile, the increased expression of CTRP9 can reduce ERs and protect the lipid deposition in Hcy induced VSMC. Furthermore, ERs can promote Hcy induced VSMC lipid deposition, inhibition of ERs can reduce Hcy induced VSMC lipid deposition, and CTRP9 may play a protective role in Hcy induced VSMC lipid deposition and foam cell transformation through negative regulation of ERs. In addition, The CTRP9 promoter in the Hcy group showed hypermethylation. At the same time as Hcy intervention, overexpression of DNMT1 increases the methylation level of the CTRP9 promoter, while 5-Azc can reduce the methylation level of the CTRP9 promoter. Finally, Hcy can up-regulate the expression of DNMT1 and down-regulate the expression of CTRP9. After overexpression of DNMT1, the expression of CTRP9 is further decreased. After 5-Azc inhibition of DNMT1, the expression of DNMT1 decreases, while the expression of CTRP9 increases. It is suggested that the molecular mechanism of Hcy inhibiting the expression of CTRP9 is related to the hypermethylation of the CTRP9 promoter induced by Hcy and regulated by DNMT1. 5-Azc can inhibit the expression of DNMT1 and reverse the regulatory effect of DNMT1 on CTRP9. Overall, the results of the present study suggested that Hcy induces DNA hypermethylation in the CTRP9 promoter region by up-regulating DNMT1 expression, and negatively regulates ERs mediated VSMC lipid deposition and foam cell formation. CTRP9 may potentially be a therapeutic target in the treatment of hyperhomocysteinemia and As.

Wnt/ß-catenin signalling pathway in breast cancer cells and its effect on reversing tumour drug resistance by alkaloids extracted from traditional Chinese medicine.

Breast cancer is a high-risk disease with a high mortality rate among women. Chemotherapy plays an important role in the treatment of breast cancer. However, chemotherapy eventually results in tumours that are resistant to drugs. In recent years, many studies have revealed that the activation of Wnt/ß-catenin signalling is crucial for the emergence and growth of breast tumours as well as the development of drug resistance. Additionally, drugs that target this pathway can reverse drug resistance in breast cancer therapy. Traditional Chinese medicine has the properties of multi-target and tenderness. Therefore, integrating traditional Chinese medicine and modern medicine into chemotherapy provides a new strategy for reversing the drug resistance of breast tumours. This paper mainly reviews the possible mechanism of Wnt/ß-catenin in promoting the process of breast tumour drug resistance, and the progress of alkaloids extracted from traditional Chinese medicine in the targeting of this pathway in order to reverse the drug resistance of breast cancer.

Occurrence and fate characteristics of isoproturon from garlic cultivation to household processing: Implication for human exposure.

The fate characteristics of isoproturon (IPU) from garlic cultivation to household processing was elucidated by a tracing UHPLC-MS/MS based on the favorable storage stability. The occurrence, pharmacokinetics dissipation and terminal magnitude of IPU were reflected by parameters including original deposition of 31-170 µg kg-1, half-lives of 11.5-19.4 d, and final concentrations of <1.0-250.6 µg kg-1. The processing factors of IPU were further clarified in terms of washing, stir-frying and pickling, with processing factors of 0.008-0.828. The chronic dietary risks (%ADI) were assessed as 1.516-5.242 %, whereas the short-term exposures from green garlic should be continuously emphasized over 99th percentile with unacceptable %ARfD of 147.144-5074.018 %. The acute and chronic risk magnitude significantly decreased by a factor 2.0-125.0 and 2.2-3.3 from raw garlic crops to processed products, respectively. What was noteworthy was the unacceptable acute risks of IPU from green garlic at 99.9th percentile even after a series of processing procedures.

Antioxidant properties of polyphenols from snow chrysanthemum (Coreopsis tinctoria) and the modulation on intestinal microflora in vitro.

CONTEXT: Coreopsis tinctoria Nutt (Asteraceae), named snow chrysanthemum, is known to have a high level of polyphenols. However, the potential prebiotic effect on modulating intestinal microflora is still unclear. OBJECTIVE: The chemical composition, antioxidant properties of snow chrysanthemum polyphenols (SCPs) and their effects on human intestinal microbiota were investigated. MATERIALS AND METHODS: SCPs were extracted using ultrasonic-assisted extraction, and further determined using UPLC-QE Orbitrap/MS. Five assays were used to investigate the antioxidant activities of SCPs. Subsequently, the effects of SCPs on intestinal microbiota in vitro were determined by high throughput sequencing and bioinformatics analysis. RESULTS: Marein, isookanin and cymaroside were the major phenolic compounds, which accounted for 42.17%, 19.53% and 12.25%, respectively. Marein exhibited higher scavenging capacities in DPPH (EC50 = 8.84 µg/mL) and super anion radical assay (EC50 = 282.1 µg/mL) compared to cymaroside and isookanin. The antioxidant capacity of cymaroside was weakest among the three phenolic compounds due to the highest EC50 values, especially for superoxide anion radical assay, EC50 > 800 µg/mL. The result of in vitro fermentation showed that the three phenolic compounds increased the relative abundances of Escherichia/Shigella, Enterococcus, Klebsiella, etc., and isookanin notably increased the relative abundance of Bifidobacterium and Lactobacillus. DISCUSSION AND CONCLUSIONS: SCPs exhibited antioxidant properties and potential prebiotic effects on modulating the gut microbiota composition. The findings indicated that SCPs consumption could exert prebiotic activity that is beneficial for human health.

Xiaoyao Pill Improves the Affective Dysregulation of Sleep-deprived Female Mice by Inhibiting Brain Injury and Regulating the Content of Monoamine Neurotransmitter.

BACKGROUND: Sleep curtailment is a serious problem in many societies. Clinical evidence has shown that sleep deprivation is associated with mood dysregulation, formation of false memory, cardio-metabolic risk factors and outcomes, inflammatory disease risk, and all-cause mortality. The affective disorder dysregulation caused by insufficient sleep has become an increasingly serious health problem. However, to date, not much attention has been paid to the mild affective dysregulation caused by insufficient sleep, and there is no clear and standard therapeutic method to treat it. The Xiaoyao Pill is a classic Chinese medicinal formula, with the effect of dispersing stagnated hepatoqi, invigorating the spleen, and nourishing the blood. Therefore, it is most commonly used to treat gynecological diseases in China. In the present study, the effects of the Xiaoyao Pill on affective dysregulation of sleep-deprived mice and its underlying molecular mechanisms were investigated. METHODS: Forty adult female mice were used in the present study. The sleep deprivation model was established by improving the multi-platform water environment method. After 7 consecutive days of sleep deprivation, the mice were administrated low (LXYP, 0.32mg/kg) and high (HXYP, 0.64 mg/kg) doses of the Xiaoyao Pill for two weeks. Then, the body weight, behavioral deficits, and histopathology were evaluated. Meanwhile, the expression of c-fos protein and the concentrations of monoamine neurotransmitters in the hippocampus and prefrontal cortex were determined after two weeks of treatment. RESULTS: Xiaoyao Pill treatment significantly increased body weight and sucrose consumption and decreased the irritability scores of the sleep-deprived mice. Meanwhile, Xiaoyao Pill treatment prevented brain injury and inhibited the expression of c-fos protein in the hippocampus and prefrontal cortex. In addition, HXYP treatment significantly upregulated the levels of NE in the hippocampus and prefrontal cortex (p < 0.01). LXYP treatment significantly up-regulated the levels of 5-HT in the prefrontal cortex. Meanwhile, both HXYP and LXYP treatment significantly upregulated the levels of DA in the prefrontal cortex (p < 0.05 or p < 0.01) of sleep-deprived mice. CONCLUSION: The present study demonstrates that Xiaoyao Pill treatment prevented the behavioral deficits of mice induced by sleep deprivation by promoting the recovery of brain tissue injury and up-regulating the levels of NE, 5-HT, and DA in the brain tissue.

3D printed double-network alginate hydrogels containing polyphosphate for bioenergetics and bone regeneration.

Low mechanical strength, poor processability, and low bioactivity of hydrogels limit their application in bone tissue engineering severely. Herein, a new 3D-printable, osteoinductive, and bioenergetic-active double-network (DN) hydrogel containing sodium alginate (SA), poly (ethylene glycol) diacrylate (PEGDA), and sodium polyphosphate (PolyP) was developed via a two-step method. The synergy of the covalent cross-linking network and the ionic cross-linking network improves the mechanical properties of the hydrogel. And the pre-gel with Ca2+ has better 3D printing performance to print complex tissue engineering scaffolds than common hydrogels. In addition, the incorporation of PolyP into DN hydrogel matrix significantly improves the bioactivity of hydrogels. The bioenergetic effect of PolyP improves adenosine triphosphate content of cells significantly to promote cell activities such as migration. The in vitro osseointegration investigation suggests that the orthophosphate monomer units, which are degradation fragments of PolyP, provide enough phosphoric acid units for the formation of calcium phosphate and accelerate the osteogenic differentiation of cells greatly. Therefore, the proposed printable, bioenergetic-active, osteoinductive DN hydrogel is potential to solve the problems of complex tissue engineering scaffolds and be applied in energy-crucial bone tissue regeneration.

[Responses of photosynthesis and P/Fe traits to P application in soybean under stress of low Fe.] / é“èƒè¿«ä¸‹å¤§è±†å ‰åˆå’Œç£·/é“æ€§çŠ¶å¯¹ç£·ç´ çš„å“åº”.

We examined the effects of phosphorus (P) levels on photosynthetic and P/Fe traits of soybean under the stress of low Fe and their genotypic differences, to provide a theoretical basis for rational application of P and Fe fertilizer. Six P-efficient and six P-inefficient soybean varieties screened in the early stage were used as experimental materials. Four treatments of P:Fe ratio were set, including 0:30, 30:30, 150:30 and 300:30 (µmol·L-1). We measured chlorophyll fluorescence traits and P-Fe utilization efficiency in soybean. A stepwise regression equation was established with seed weight per plant. Pathway analysis was performed, with the response of P-efficient and P-inefficient soybean genotypes to different P:Fe treatments being comprehensively evaluated by factor scores. The results showed significant main and interactive effects of genotype and P:Fe on the relative electron transfer rate of photosystem Ⅱ (ETR) at beginning of flowering stage (R1), the proportion of the energy absorbed by photosystem Ⅱ dissipated into heat (NPQ) at R1 stage, and proportion of energy absorbed by photosystem Ⅱ devoted to the photochemical reaction (qL) at R1 stage. Results of canonical correlation analysis showed a negative correlation between P utilization efficiency of seed at full maturity stage (R8) and photosynthetic rate at R1 stage of P-efficient genotypes. Seed Fe utilization efficiency of P-inefficient genotypes at R8 stage was positively correlated with NPQ at R1 stage, but negatively correlated with qL at R1 stage. The actual photochemical efficiency of PSⅡ (ΦPSⅡ) at R1 stage was negatively correlated with P-efficient genotypes, but positively correlated with P-inefficient genotypes, which indicated that ΦPSⅡ at R1 stage was an important indicator for identifying soybean genotypes with different P efficiency under stress of low Fe. The comprehensive performance of P-efficient soybean genotypes decreased first and then increased with P level, while P-inefficient soybean genotypes increased first and then decreased. The inflection point of both genotypes appeared in P:Fe of 30:30. Thus, P:Fe ratio of 30:30 could be used as a threshold to identify soybean genotypes with different P efficiency under stress of low Fe. In conclusion, P fertilizer application should be equal to or greater than 1:1 (P:Fe) when planting P-efficient soybean genotypes in low Fe area, while P fertilizer application should not exceed 1:1 (P:Fe) when planting P-inefficient soybean genotypes.

Lycium barbarum polysaccharide protects against Homocysteine-induced Vascular smooth muscle cell proliferation and phenotypic transformation via PI3K/Akt pathway.

Lycium barbarum polysaccharide (LBP) is an alkaloid extracted from lycium barbarum. LBP is the active component of lycium barbarum used to treat hypertension, atherosclerosis and other cardiovascular diseases in Chinese traditional medicine. However, the underlying cellular and molecular mechanisms of LBP- mediated activity in vascular disease remain poorly understood. In the present study, we showed the protective effect of LBP in vascular smooth muscle cells. Our results indicate that LBP significantly reduces the proliferation of VSMCs caused by Homocysteine (Hcy) and inhibits the phenotypic transformation of VSMCs caused by Hcy, from contractile to synthetic. LBP inhibited the protein expression of PI3K and Akt caused by Hcy, and increased the expression of miR-145. The results indicate that LBP exhibits substantial therapeutic potential for the treatment of Hcy-induced VSMCs proliferation and phenotypic transformation through inhibition of PI3K/Akt signaling pathway.

Exploration of natural phosphatidylcholine sources from six beans by UHPLC-Q-HRMS.

Beans are a rich source of phosphatidylcholine (PC). This study aims to explore natural PC sources rich in polyunsaturated fatty acid (PUFA) with nutritional interest. PCs from six beans were purified (purity > 98.2%) by thin layer chromatography (TLC), and subsequently identified by ultra-high performance liquid chromatography-Quadrupole (Q)-high-resolution mass spectrometry (UHPLC-Q-HRMS). Results showed that the PC content of chickpea (Cicer arietinum) and soybean (Glycine max) was 50.0 and 34.0 mg/g, respectively, which was significantly higher than that of other beans (P < 0.05). Gas chromatographic analysis showed that soybean contained high proportion of PUFA (58.78%), and chickpea contained high proportion of docosahexaenoic acid (DHA) (2.73%). A total of 49 molecular species were identified by UHPLC-Q-HRMS. (18:2-18:2)PC was predominant in soybean, adzuki bean, runner bean, and common bean. (16:0-18:1)PC was the major species of chickpea PC, and many ether PC species and DHA-PC were identified. Discriminatory analysis by principal component analysis (PCA) indicated that the molecular profiles of chickpea PC were significantly different from other beans studied. The findings suggest that chickpea appears to be an interesting plant source of DHA and ether lipids for dietary supplement. PRACTICAL APPLICATION: In this study, we reported an UHPLC-Q-HRMS technique to identify PC molecular species of six beans. The diversity of PC molecular species in the different beans was classified using chemometrics. This analytical method not only provides comprehensive information to nutritionists about the PC distribution in different beans, but also can identify biomarkers for bean flour fraud identification in food supplementation. Furthermore, the approach gives fragmentation patterns of several PC species and could be further applied to determine the chemical structure of PC molecular species from many natural resources.

Folate Protects Hepatocytes of Hyperhomocysteinemia Mice From Apoptosis via Cystic Fibrosis Transmembrane Conductance Regulator (CFTR)-Activated Endoplasmic Reticulum Stress.

Folate deficiency is a known risk factor for liver injury; however, the underlying mechanism remains unclear. In this study, we employed a high homocysteine-induced liver injury model of Apolipoprotein E-deficient (ApoE-/- ) mice fed high-methionine diet and found that high homocysteine induced endoplasmic reticulum (ER) stress and liver cell apoptosis by downregulation of cystic fibrosis transmembrane conductance regulator (CFTR) expression; observations that were attenuated with supplementation of dietary folate. The regulation on CFTR expression was mediated by CFTR promoter methylation and trimethylation of lysine 27 on histone H3 (H3K27me3). Mechanistically, folate inhibited homocysteine-induced CFTR promoter methylation and H3K27me3, which resulted in upregulation of CFTR expression, and reduced ER stress and liver cell apoptosis. Further study showed that folate inhibited the expression of DNA methyltransferase 1 and enhancer of zeste homolog 2, downregulated the cellular concentrations of S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) and upregulated the SAM/SAH ratio, leading to the inhibition of Hcy-induced DNA hypermethylation and H3K27me3 in CFTR promoter. In conclusion, our results provide insight into the protective role of folate in homocysteine-induced ER stress and liver cell apoptosis through the regulation of CFTR expression. J. Cell. Biochem. 118: 2921-2932, 2017. © 2017 Wiley Periodicals, Inc. HIGHLIGHTS: Folate protects hepatocytes of hyperhomocysteinemia mice from apoptosis. Folate alleviates Hcy-induced hepatocyte apoptosis. Folate inhibits Hcy-induced ER stress via upregulation of CFTR expression in hepatocytes. Folate inhibits Hcy-induced methylation of CFTR promotor and H3K27me3.

Antiandrogen Therapy with Hydroxyflutamide or Androgen Receptor Degradation Enhancer ASC-J9 Enhances BCG Efficacy to Better Suppress Bladder Cancer Progression.

Recent studies suggest that the androgen receptor (AR) might play important roles in influencing bladder cancer progression, yet its clinical application remains unclear. Here, we developed a new combined therapy with Bacillus Calmette-Guérin (BCG) and the AR degradation enhancer ASC-J9 or antiandrogen hydroxyflutamide (HF) to better suppress bladder cancer progression. Mechanism dissection revealed that ASC-J9 treatment enhanced BCG efficacy to suppress bladder cancer cell proliferation via increasing the recruitment of monocytes/macrophages that involved the promotion of BCG attachment/internalization to the bladder cancer cells through increased integrin-α5ß1 expression and IL6 release. Such consequences might then enhance BCG-induced bladder cancer cell death via increased TNFα release. Interestingly, we also found that ASC-J9 treatment could directly promote BCG-induced HMGB1 release to enhance the BCG cytotoxic effects for suppression of bladder cancer cell growth. In vivo approaches also concluded that ASC-J9 could enhance the efficacy of BCG to better suppress bladder cancer progression in BBN-induced bladder cancer mouse models. Together, these results suggest that the newly developed therapy combining BCG plus ASC-J9 may become a novel therapy to better suppress bladder cancer progress.

[Effect of oxymatrine on JAK2/STAT3 signaling in renal tissues of rats with septic shock].

OBJECTIVE: To explore the effect of oxymatrine (OMT) on JAK2/STAT3 signaling in renal tissues of rats with septic shock. METHOD: The cecal ligation and puncture (CLP) was adopted to establish the rat septic shock model. Fifty-six male SD rats were randomly divided into 7 groups: the sham operation group, the model (CLP) group, CLP + OMT high, middle, low-dose (52, 26, 13 mg x kg(-1), vena caudalis bolus) groups and the positive control (CLP + dexamethasone, 10 mg x kg(-1)) group. The pathological changes in renal tissues were examined with lightmicroscope. BUN content was determined by urine enzymatic method. Expressions of tumournecrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) mRNA in renal tissues were determined by RT-PCR. Expression of JAK2 and STAT3 in renal tissues determined by Western blot. Changes in tumournecrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) contents in renal tissue were determined by radioimmunoassay. RESULT: OMT of different doses could inhibit the JAK2 and STAT3 activation in renal tissues (P<0.05), and decrease the protein expression of JAK2, STAT3, TNF-alpha and IL-1beta mRNA (P<0.05). Besides, it could reduce TNF-alpha and IL-1beta contents in renal tissue homogenate (P<0.05), serum BUN content (P<0.05), and improve such lesions as tissue hyperemia, edema and inflammatory cell infiltration, with identical results in medium and high-dose OMT groups, and the positive control group. CONCLUSION: OMT can inhibit JAK2/STAT3 signaling activity to reduce the expression of proin-flammatory factors (TNF-alpha, IL-1beta) and treat the renal injury in rats with septic shock.

Oxymatrine protects against myocardial injury via inhibition of JAK2/STAT3 signaling in rat septic shock.

Oxymatrine (OMT), an alkaloid extracted from Sophora japonica (kushen), is used to treat inflammatory diseases and various types of cancer in traditional Chinese medicine. However, the cellular and molecular mechanisms underlying the anti­inflammatory activity of OMT remain poorly understood. The present study explored the protective effect of OMT on myocardial injury in rats with septic shock by inhibiting the activation of the janus kinase­signal transducer and activator of transcription (JAK/STAT) signaling pathway. OMT treatment was found to significantly inhibit the activation of JAK2 and STAT3 in myocardial tissue. It also attenuated the expression of pro­inflammatory cytokines, including interleukin­1ß and tumor necrosis factor­α. In addition, OMT exhibited anti­inflammatory properties as heart function and myocardial contractility was improved and pathological and ultrastructural injury was prevented in myocardial tissue induced by septic shock. The results indicate that OMT exhibits substantial therapeutic potential for the treatment of septic shock­induced myocardial injury through inhibition of the JAK2/STAT3 signaling pathway.

[Effect of oxymatrine on vascular calcification of humans umbilical vein smooth muscle cells and its underlying mechanism].

OBJECTIVE: To observe the effect of oxymatrine (OMT) on calcification of humans umbilical vein smooth muscle cells and its underlying mechanism. METHOD: Human umbilical vein smooth muscle cells (HUSMCs) were calcified by beta-giycerophos-phosphate (beta-GP) and then divided into 6 groups: the control group, the calcification group, the pure OMT group, and lower, middle and higher-dosage OMT groups. Cell calcification were observed by Von Kossa staining, calcium content in HUSMCs were determined by the colorimetric method, the alkaline phosphatase (ALP) activity in HUSMCs were determined by phenyl diphosphate-2-sodium, the osteocalcin (OC) level in HUSMCs were determined by radioimmunossay, the transforming growth factor-beta1 (TGF-beta1) level in HUSMC culture medium and the content changes in psmad2/3 and smad2/3 were determined by the ELISA method, and the expression of Core binding factor alpha1 (Cbfalpha1) protein in HUSMCs were determined by western blot method. RESULT: Compared with the control group, the calcification group showed a great number of black granules among the smooth muscle cells and significant increase in the content of calcium and OC and the activity of ALP; OMT intervention can decrease the content of calcium, OC, TGF-beta1, psmad2/3 and Cbfalpha1 and the activity of ALP. And high-dosage OMT group had better effect than middle and low-dosage groups. CONCLUSION: OMT can effectively inhibit beta-GP-induced HUSMC calcification and its effect on reducing TGF-beta1, psmad2/3 and Cbfalpha1 may be one of its mechanisms in inhibiting HVSMC calcification.

[Effect of oxymatrine on JAK/STAT iteral in rat lung tissue with sepsis].

OBJECTIVE: To explore the effects of oxymatrine (OMT) on JAK/STAT iteral in rat lung tissue with sepsis. METHOD: Fifty-six male SD rats were randomly divided into 6 groups: sham operation group, model (CLP) group, CLP + OMT high, middle, low-dose groups (52, 26, 13 mg x kg(-1), vena caudalis bolus), and positive control group (dexamethasone, 10 mg x kg(-1), vena caudalis bolus) to observe the effects of oxymatrine on the ratio between wet weight of the lung and dry weight of the lung (W/D) and pulmonary coefficient, gross changes and pathological changes examined with lightmicroscope in the pulmonary tissue. Changes in JAK2 and STAT3 activity in the pulmonary tissue were determined by immunohistochemical method. Tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) levels in pulmonary tissue were determined by radioimmunoassay. RESULT: OMT could decrease significantly the JAK2 and STAT3 positive reaction and activity in the pulmonary tissue (P < 0.05). TNF-alpha and IL-6 levels in pulmonary tissue homogenate decreased markedly (TNF-alpha decreased 36%, 26%, 16% and IL-6 decreased 46%, 39%, 24% on CLP + OMT 52, 26 mg x kg(-1) and 13 mg x kg(-1) groups. P < 0.05 or P < 0.01). OMT could decrease the ratio between wet weight of the lung and dry weight of the lung and the pulmonary coefficient, improve the condition of pulmonary hyperemia, edema, infiltrate of heterophil granulocyte and emerge of asphyxial membrane, and alleviate the inflammatory reaction. And the results were equal to those of the positive control (CLP + dexamethasone) group. CONCLUSION: OMT can inhibit JAK/STAT iteral activity and reduce the expression of proinflammatory factor (TNF-alpha, IL-6) and antagonize the lung injury in a rat model of sepsis.

[Effect of oxymatrine on NF-kappaB and other cell factors in rats lung tissue with septic shock].

OBJECTIVE: To explore the effects of oxymatrine (OMT) on NF-kappaB and other cell factors in rat lung tissue with septic shock. METHOD: Fifty-six male SD rats were randomly divided into 7 groups: sham operation group, OMT control group, model (CLP) group, CLP + OMT high, middle, low-dose group, positive control group. Changes in NF-kappaB (p65) and IkB-alpha activity in the pulmonary tissue were determined by immunohistochemical method, tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) levels in pulmonary tissue were determined by radioimmunoassay. RESULT: OMT could decrease significantly the NF-kappaB (p65) and IkB-alpha activity in the pulmonary tissue (P < 0.05), TNF-alpha and IL-6 levels in pulmonary tissue homogenate decreased markedly (P < 0.05). OMT could elevate the content of PaO2, SaO2, decrease the content of PaCO2, HCO3- and decrease the ratio between wet weight of the lung and dry weight of the lung and the PWI. CONCLUSION: OMT can inhibit NF-kappaB-inducing kinase (NIK), NF-kappaB activity and reduce the expression of proinflammatory factor (TNF-alpha, IL-6) and antagonize the lung injury in a rat model of septic shock.