RÉSUMÉ
BACKGROUND: Pulmonary fibrosis (PF) is a progressive and severe respiratory disease for which there is still a lack of satisfactory treatment methods other than lung transplantation. Evening primrose (EP) is widely used in Chinese folk medicinal herbs, especially for the treatment of lung-related diseases. However, the protective effect of evening primrose against PF has yet to be reported. PURPOSE: This study explores the pharmacological effect of EP and its possible active components against PF from the perspectives of lung function, histopathological staining, and molecular biology assays. METHODS: Establishing a rat pulmonary fibrosis model using bleomycin to detect lung function, pathological changes, and collagen deposition. TGF-ß1 was used to establish an in vitro model of PF in BEAS-2B cells, and the active ingredients in evening primrose were screened. Then, the therapeutic effects of 1-Oxohederagenin (C1) and remangilone C (C2) derived from EP were observed in an in vivo model of bleomycin-induced PF, and the differentially expressed genes between the C1 and C2 treatment groups and the model group were screened with transcriptome sequencing. Finally, TGF-ß1-induced damage to HFL1 cell was used to explore the specific mechanisms by which C1 and C2 alleviate PF and the involvement of ß-catenin signaling. RESULTS: Evening primrose extract showed some ameliorative effects on bleomycin-induced PF in rats, manifested as reduced pathological damage and reduced collagen deposition. The chemical components of C1 and C2 potently ameliorated BLM-induced PF in animals and effectively inhibited fibroblast activation by interfering with ß-catenin signaling. CONCLUSION: Evening primrose extract has certain ameliorative effects on PF. In addation, C1 and C2 might be related with the suppression of fibroblast activation by inhibiting ß-catenin signaling.
RÉSUMÉ
BACKGROUND: Pulmonary fibrosis (PF) is an end-stage change in many interstitial lung diseases, whereas no proven effective anti-pulmonary fibrotic treatments. Forsythoside A (FA) derived from Forsythia suspensa (Thunb.) Vahl, has been found to possess lung-protective effect. However, studies on its anti-pulmonary fibrosis effect are limited and its mechanism of action remains unknown. PURPOSE: This study aimed to explore the underlying mechanism of FA on PF. METHODS: Male C57BL/6 mice were randomized into normal (CON), model (BLM), pirfenidone (PFD), low- and high-dose FA (FA-L, FA-H, respectively). Except for the CON group, which was injected with the same dose of saline, the model of PF was established by intratracheal instillation of BLM, during which the survival rate and body weight changes of the mice were measured. The lung histopathology was evaluated by Hematoxylin-eosin, Sirius red, and Masson staining. Transcriptome analysis was performed to screen for the differential genes associated with the role of FA in PF. Differential genes in normal and pulmonary fibrosis patients with the GSE2052 dataset were analyzed in the GEO database. The levels of CTGF, α-SMA, MMP-8 in lung and TNF-α in bronchoalveolar lavage fluid (BALF) were detected by ELISA. The levels of HYP in lungs were detected by digestion. The mRNA and protein levels of MMP-7, E-cadherin, CD31, α-SMA, TGF-ß1, IL-6, ß-catenin, ZO-1, PTPRB, E-cadherin, and vimentin in lungs were detected by RT-qPCR and Western blot. The expression of CD31, α-SMA, TGF-ß1 and ZO-1 were detected by immunofluorescence. TGF-ß1-stimulated HFL1 cells and human umbilical vein endothelial cells (HUVECs) were used in an attempt to explore the possible role of protein tyrosine phosphatase receptor type B (PTPRB) involved in FA-induced improvement of PF. RESULTS: The results showed that FA could improve the survival rate and body weight of PF mice. FA could alleviate the symptoms of alveolar wall thickening, inflammatory cell infiltration, blue collagen fiber deposition, collagen fiber type â and type â ¢ in mice with PF. In addition, FA could reduce the levels of HYP, CTGF, α-SMA, TGF-ß1, TNF-α, ß-catenin and MMP8, and regulate the expression levels of CD31, ZO-1, PTPRB and E-cadherin in lung of mice with PF, inhibiting endothelial-to-mesenchymal transition (EndMT) and fibroblasts proliferation. In the GSE2052 dataset, the expression level of PTPRB is reduced in lung tissue from PF patients, and results from transcriptome sequencing indicate that PTPRB expression is also reduced in PF mice. In addition, the effect of FA on TGF-ß1-induced HFL1 or HUVECs cells could be attenuated by the inhibitor of PTPRB, suggesting that the effect of FA on PF is related to PTPRB. CONCLUSION: This study demonstrated that FA could ameliorate PF by inhibiting lung fibroblast proliferation and EndMT, and that PTPRB might be a target of FA to ameliorate PF, which provided evidence to support FA as a candidate phytochemical for PF.
Sujet(s)
Forsythia , Hétérosides , Poumon , Souris de lignée C57BL , Fibrose pulmonaire , Transduction du signal , Animaux , Mâle , Fibrose pulmonaire/traitement médicamenteux , Transduction du signal/effets des médicaments et des substances chimiques , Poumon/effets des médicaments et des substances chimiques , Poumon/anatomopathologie , Hétérosides/pharmacologie , Forsythia/composition chimique , Transition épithélio-mésenchymateuse/effets des médicaments et des substances chimiques , Souris , Prolifération cellulaire/effets des médicaments et des substances chimiques , Fibroblastes/effets des médicaments et des substances chimiques , Receptor-Like Protein Tyrosine Phosphatases, Class 3/métabolisme , Humains , Cellules endothéliales de la veine ombilicale humaine/effets des médicaments et des substances chimiques , Modèles animaux de maladie humaine , Actines/métabolisme , BléomycineRÉSUMÉ
BACKGROUND: Modern pharmacological studies show that Epimedium sagittatum Maxim (EPI) has antioxidant, antiapoptotic, anti-inflammatory effects. However, the effects of EPI on adriamycin-induced nephropathy are unclear. AIM: The main purpose of this study is to investigate the effects of EPI on adriamycin-induced nephropathy in rats. METHODS: The chemical composition of EPI was detected by high performance liquid chromatography. Network pharmacology was used to collect the effects of EPI on adriamycin nephropathy; renal histological changes, podocyte injury, inflammatory factors, oxidative stress levels, apoptosis levels, and the PI3K/AKT signaling pathway were examined. Moreover, analyze the effects of icariin (the representative component of EPI) on adriamycin-induced apoptosis and PI3K/AKT signaling pathway of NRK-52e cells. RESULTS: Network pharmacological results suggested that EPI may ameliorate adriamycin-induced nephropathy by inhibiting inflammatory response and regulating the PI3K/AKT signaling pathway. The experimental results showed that EPI could improve pathological injury, renal function, podocyte injury, and inhibit inflammation, oxidative stress, apoptosis in adriamycin-induced nephropathy rats through the PI3K/AKT signaling pathway. Furthermore, icariin inhibited adriamycin-induced mitochondrial apoptosis in NRK-52e cells. CONCLUSION: This study suggested that EPI ameliorates adriamycin-induced nephropathy by reducing inflammation and apoptosis through the PI3K/AKT signaling pathway, icariin may be the pharmacodynamic substance basis for this effect.
Sujet(s)
Epimedium , Animaux , Rats , Phosphatidylinositol 3-kinases , Protéines proto-oncogènes c-akt , Apoptose , Doxorubicine/toxicité , Inflammation/induit chimiquement , Inflammation/traitement médicamenteux , Transduction du signalRÉSUMÉ
BACKGROUND: Severe inflammation of the lungs results from acute lung injury (ALI), a common life-threatening lung disease with a high mortality rate. The ligand-activated transcription factor peroxisome proliferator-activated receptor (PPAR) γ plays essential roles in diverse biological processes including inflammation, metabolism, development, and immune response. Salvianolactone acid A (SA) is a terpenoid derived from the herb Salvia miltiorrhiza. However, there is a scarcity of experimental evidence indicating whether the effect of SA on ALI occurs via PPAR-γ. METHODS: SA (20 or 40 mg/kg, i.g., 1 time/day) was administered to mice for 3 d, followed by the induction of ALI by intranasal lipopolysaccharide (LPS, 10 mg/kg). The lung function and levels of inflammation, reactive oxygen species (ROS), immune cells, apoptosis, and PPAR-γ were examined. The antagonistic activity of GW9662 (GW, 1 µM, specific PPAR-γ blocker) and PPAR-γ transfection silencing against SA (10 µM) in BEAS-2B cells induced by LPS (10 µg/ml, 24 h) was also investigated to assess whether the observed effects caused by SA were mediated by PPAR-γ. RESULTS: The results showed that lung histopathological injury, the B-line, the fluorescence intensity of live small animal, and the biomarkers in BALF or lung in the treatment of SA could regulate significantly. In addition, SA obviously decreased the levels of ROS and apoptosis in the primary lung cells, and MDA, increased the levels of GSH-Px and SOD. SA reduced levels of macrophages and neutrophils. Furthermore, SA reduced the protein levels of Keap-1, Cleaved-caspase-3, Cleaved-caspase-9, p-p65/p65, NLRP3, IL-1ß, and upregulated the levels of p-Nrf2/Nrf2, HO-1, Bcl-2/Bax, PPAR-γ, p-AMPK/AMPK in lung tissue. In addition, silencing and inhibition of PPAR-γ effectively decreased the protective effects of SA in BEAS-2B cells induced by LPS, which might indicate that the active molecules of SA regulate ALI via mediation by PPAR-γ, which exhibited that the effect of SA related to PPAR-γ. CONCLUSIONS: The anti-ALI effects of SA were partially mediated through PPAR-γ signaling. These data provide the molecular justification for the usage of SA in treating ALI and can assist in increasing the comprehensive utilization rate of Salvia miltiorrhiza.
Sujet(s)
Lésion pulmonaire aigüe , Salvia miltiorrhiza , AMP-Activated Protein Kinases , Animaux , Inflammation , Lipopolysaccharides , Poumon , Souris , Facteur-2 apparenté à NF-E2 , Facteur de transcription NF-kappa B , Récepteur PPAR gamma , Espèces réactives de l'oxygèneRÉSUMÉ
PURPOSE: To investigate the effect and molecular mechanism of miR-490 in prostate cancer. METHODS: qRT-PCR was used to detect the expression of miR-490 in human prostate cancer PC-3 cell line (the control group B) and normal human prostate cells (the control group A), the overexpression vector (the positive group) and human prostate cancer PC-3 cells transfected by negative control (the negative group) were built, and the results of the transfection were detected by qRT-PCR. RESULTS: The relative expression levels of miR-490 in the cells in the control group A, the control group B and the negative group were lower than that in the positive group (p<0.05). The OD values of the cells in the negative and positive groups both increased continuously over time (p<0.05). The OD values of the cells in the positive group at each time point were lower than those in the negative group (p<0.05). CONCLUSION: The low expression and overexpression of miR-490 in prostate cancer may inhibit the proliferation, migration and invasion of prostate cancer cells and promote apoptosis by inhibiting the phosphorylation of AKT signal pathway.
Sujet(s)
microARN/métabolisme , Tumeurs de la prostate/génétique , Lignée cellulaire tumorale , Humains , Mâle , Cellules PC-3 , TransfectionRÉSUMÉ
Prostate cancer (PC) is the most common cancer in men; however, limited effect is obtained due to the therapy resistance. CASC2 acts as a tumor suppressor in human malignancies serving as a ceRNA for miRNAs; Sprouty2 (SPRY2), a key antagonist of RTK signaling, also serves as a tumor suppressor. Herein, CASC2 and SPRY2 expression was down-regulated in PC tissues and cell lines; the overexpression of CASC2 and SPRY2 could suppress PC cell proliferation, promote PC cell apoptosis, and enhance the sensitivity of PC cells to docetaxel. CASC2 positively regulated SPRY2 expression and inhibited downstream extracellular regulated protein kinases (ERK) signaling activation through SPRY2. By using online tools, miR-183 might be a direct target of CASC2, and might simultaneously bind to the 3'UTR of SPRY2. The direct binding between CASC2, miR-183 and SPRY2 was then validated; miR-183 inhibition enhanced the cytotoxicity of docetaxel on PC cells, which could be partially attenuated by SPRY2 knockdown. In summary, CASC2 competes with SPRY2 for miR-183 binding to rescue the expression of SPRY2 in PC cells, thus enhancing the sensitivity of PC cells to docetaxel through SPRY2 downstream ERK signaling pathway; CASC2 and SPRY2 might be novel adjuvants for docetaxel-based chemotherapy for PC.
Sujet(s)
Antinéoplasiques/pharmacologie , Docetaxel/pharmacologie , Protéines et peptides de signalisation intracellulaire/génétique , Protéines membranaires/génétique , microARN/génétique , Tumeurs de la prostate/anatomopathologie , ARN long non codant/génétique , ARN/génétique , Protéines suppresseurs de tumeurs/génétique , Lignée cellulaire tumorale , Résistance aux médicaments antinéoplasiques/génétique , Techniques de knock-down de gènes , Humains , Mâle , Tumeurs de la prostate/génétiqueRÉSUMÉ
Bladder cancer is the most common malignant tumor of the urinary system, and it is also an important cause of death by cancer globally. Increasing number of studies have shown that miRNAs can be used as prognostic markers for cancers. This study made use of the data available in the Cancer Genome Atlas in order to statistically analyze reported expression levels of miRNAs in samples from bladder urothelial carcinoma patients. Clinical features from a total of 399 patients and the expression data for 1,581 kinds of miRNA were included in the study. Single factor Cox regression analysis was used to identify miRNAs related to survival times for the patients. Then, through multifactors Cox regression we sorted out the independent prognostic miRNAs for the carcinoma. According to our results, 19 miRNAs were closely related to the survival times of patients with bladder urothelial carcinoma, and 3 miRNAs including hsa-mir-518b (p=0.02), hsa-mir-192 (p=0.04) and hsa-mir-7705 (p=0.04) should be useful as independent prognostic factors in patients. In addition, the survival time of those expressing high levels of hsa-mir-7705 and hsa-mir-192 was less than the survival time of those with low expression levels. However, there were no obvious differences in the survival times between high and low expressors of hsa-mir-518b. According to our results, hsa-mir-7705, hsa-mir-192 and hsa-mir-518b can be applied as independent prognostic markers for bladder urothelial carcinoma.