| Objective:Previous studies have found that curcumolol has an effect on the ultrastructure and function of sinusoidal endothelial cells,which can cause the basement membrane formation and the loss of fenestration,leading to liver microcirculatory disorders.However,the changes which in these cytokines and cell products to sinusoidal endothelial cells in the extent of structural and functional influences and the mode of action are not clear.In this study,we selected the related proteins of the signaling pathway of hepatic sinusoidal endothelial cells as a detection index to observe the secretion behavior during cell culture incubation,and inder to reveal the relationship between the changes of liver fibrosis lesions and the number of proteins related to sinusoidal endothelial cell signaling pathway.To investigate the pathogenesis of hepatic fibrosis.Methods:Construct a liver fibrosis cell model and group the cell experiments:The sinusoidal endothelial cells cultured to logarithmic growth phase were divided into groups:blank control group;Leptin activation model control group(Leptin was added to the culture medium at a final concentration of 100 μg/L for activation of hepatic sinusoidal endothelial cells);The curcumol treatment group(first activated with Leptin,the final concentration of curcumol was 50mg/L).The control group(Leptin activated cells,the final concentration of 6.25μg/mL)was treated with salvianolic acid B.(Leptin activates the cells at a final concentration of 10-6 mmol/L).The effect of curcumol on hepatic sinusoidal endothelial cells and the effect on the core expression factor of TLR4 pathway was studied from the whole cell model.To observe and compare the ultrastructural structure of hepatic sinusoidal cells,to judge and compare the degree and difference of liver fibrosis and the dynamic effects of drug action;Real-time fluorescent PCR was used to detect the mRNA expression levels of TLR4 signaling pathways(TLR4,MyD88,NF-KBp65).Western blot analysis was used to detect the expression of TLR4 signaling pathway(TLR4,MyD88,NF-KBp65).And the expression levels of p-p65,p-ERK,p-JNK,and p-P38.The experimental data was analyzed by SPSS22.0 statistical softwareResults:1.The in vitro experimental study on the improvement of LSEC ultrastructural lesions by zedoary alcohol showed that the HSEC induced by leptin was significantly reduced or even disappeared by scanning electron microscopy and transmission electron microscopy.The diameter of the window pores became smaller and the window was disappeared.After treatment with curcumol,compared with the leptin-activated model group,the number of window pores increased significantly and the window pores became larger2.The results of real-time quantitative PCR showed that ①the expression of TLR4,MyD88 and NF-κBp65 in the leptin model group was significantly higher than that in the blank control group,and the difference was statistically significant(P<0.05).②The expression levels of TLR4,MyD88 and NF-KBp65 in the colchicine group were significantly higher than those in the blank control group,and the difference was statistically significant(P<0.05).③The expression levels of TLR4,MyD88 and NF-κBp65 in salvianolic acid B group were significantly lower than those in the model group,and the difference was statistically significant(P<0.05).④The expression levels of TLR4,MyD88 and NF-KBp65 in the curcumol group were significantly lower than those in the model group,and the difference was statistically significant(P<0.05)3.The results of immunoblot assay showed that ①the expression of TLR4,MyD88 and NF-κBp65 in the leptin model group were significantly higher than that in the blank control group,and the difference was statistically significant(P<0.05).②The expression levels of TLR4,MyD88 and NF-κBp65 in the colchicine group were significantly higher than those in the blank control group,and the difference was statistically significant(P<0.05).③The expression levels of TLR4,MyD88 and NF-κBp65 in salvianolic acid B group were significantly lower than those in the model group,and the difference was statistically significant(P<0.05).④The expression levels of TLR4,MyD88 and NF-κBp65 in the curcumol group were significantly lower than those in the model group,and the difference was statistically significant(P<0.05).Conclusion:1.Curcumol has obviously effects on the ultrastructural and secretory functions of HSEC in liver fibrosis,and has a certain protective effect on liver pathological damage.2.Curcumol inhibits the expression level of the TLR4 pathway core expression factor secreted by HSEC and regulates the TLR4 signaling pathway,thereby improving liver fibrosis.3.Curcumol also has a regulatory effect on targets such asp-P38,p-ERK,and p-JNK secreted by HSEC.It can inhibit the expression level of the core factor of this pathway,and then from the new perspective of HSEC structure and secretion function changes and their relationship,the molecular mechanism of curcumol in the treatment of liver fibrosis was elucidated.Provide new ideas for the study of curcumol against liver fibrosis. |
PDF Full Text Request