| Backgrounds:Currently,about 800 million people worldwide are affected by chronic liver disease.Chronic liver disease has become one of the major public health concerns worldwide.It is worth noting that the various causes of the pathology of chronic liver disease related to chronic parenchyma injury,inflammation,fibrosis and wound healing response activation and liver fibrosis is the most important,and liver fibrosis has also become a variety of important pathological features of chronic liver disease,the mechanism involving multiple signaling pathways,the main pathological changes of intrahepatic diffuse excessive extracellular matrix deposition,extracellular matrix degradation caused by imbalance,the mechanism of hepatic stellate cell activation and proliferation of first,produce a large amount of extracellular matrix.A large number of studies have confirmed that a variety of cytokines can activate hepatic stellate cells.Platelet-derived growth factor(PDGF)is one of the effective mitotic proliferating cytokines in hepatic stellate cells,which plays a key role in the proliferation,migration,chemotaxis and other factors of hepatic stellate cells.Melatonin is a natural antioxidant that has been shown to protect against a variety of liver fibrosis,but the mechanism of action is unclear.The aim of this paper is to discuss the effect of melatonin on the activation of hepatic stellate cells and the mechanism.Methods:In this study,melatonin with concentration gradients of 1nmol/L,1?mol/L and 0.1mmol/L was used to investigate the effect of melatonin on the activation of rat HSC-T6 cells induced by PDGF(10ng/ml).After 24 h of cell culture,the cells were divided into 6 groups according to the following methods:(1)control group: PBS(2)model group: PDGF of 10ng/ml.(3)low melatonin concentration group: 10ng/ml of PDGF and 1nmol/L of melatonin(4)melatonin concentration group :10ng/ml of PDGF and 1umol/L of melatonin(5)melatonin concentration group :10ng/ml of PDGF and 0.1mmol/L of melatonin(6)inhibitor group: PDGF of 10ng/ml and AG490 of equal volume.After incubation for 48 h,the proliferation of cells in each group was detected by MTT,and P-JAK2 and P-STAT3 proteins in each group were detected by immunohistochemistry and Western Blot.Results:1.MTT experiment results showed that OD value in the model group was significantly higher than that in the control group(P<0.01).OD values of the low,medium and high melatonin concentration group and the inhibitor group showed a downward trend compared with the model group(P<0.01),and the downward trend was more obvious with the increase of melatonin concentration.2.The results of cellular immunohistochemical experiments showed that P-JAK2 protein was mainly expressed in the cytoplasm,while P-STAT3 protein was mainly expressed in the nucleus.Compared with the control group,the expression levels of P-JAK2 and P-STAT3 in the model group were significantly increased(P<0.01).Compared with the model group,the protein expressions of P-JAK2 and P-STAT3 in each melatonin dose group and inhibitor group were significantly decreased(P<0.05),and the higher the melatonin concentration was,the lower the expression levels of P-JAK2 and P-STAT3 in cells were.3.Western Blot results showed that,compared with the control group,the expression levels of P-JAK2 and P-STAT3 in HSC-T6 cells in the model group were enhanced(P<0.01).Compared with the model group,the protein expression levels of P-JAK2 and P-STAT3 in the low,medium and high dose melatonin group and the inhibitor group were decreased(P<0.05),and the protein expression level gradually decreased with the increase of melatonin concentration.Conclusion:Melatonin inhibits pdgf-induced activation of hepatic stellate cells,which is associated with the inhibition of the JAK2/STAT3 signaling pathway?... |
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