AIM:To reveal the functions of micro RNAs(mi RNAs) with respect to hepatic stellate cells(HSCs) in response to portal hypertension.METHODS:Primary rat HSCs were exposed to static water pressure(10 mm Hg,1 h) and the p...AIM:To reveal the functions of micro RNAs(mi RNAs) with respect to hepatic stellate cells(HSCs) in response to portal hypertension.METHODS:Primary rat HSCs were exposed to static water pressure(10 mm Hg,1 h) and the pressureinduced mi RNA expression profile was detected by next-generation sequencing. Quantitative real-time polymerase chain reaction was used to verify the expression of mi RNAs. A potential target of Mi R-9a-5p was measured by a luciferase reporter assay and Western blot. CCK-8 assay and Transwell assay were used to detect the proliferation and migration of HSCs under pressure.RESULTS:According to the profile,the expression of mi R-9a-5p was further confirmed to be significantly increased after pressure overload in HSCs(3.70 ± 0.61 vs 0.97 ± 0.15,P = 0.0226),which resulted in the proliferation,migration and activation of HSCs. In vivo,the up-regulation of mi R-9a-5p(2.09 ± 0.91 vs 4.27 ± 1.74,P = 0.0025) and the down-regulation of Sirt1(2.41 ± 0.51 vs 1.13 ± 0.11,P = 0.0006) were observed in rat fibrotic liver with portal hypertension. Sirt1 was a potential target gene of mi R-9a-5p. Through restoringthe expression of Sirt1 in mi R-9a-5p transfected HSCs on pressure overload,we found that overexpression of Sirt1 could partially abrogate the mi R-9a-5p mediated suppression of the proliferation,migration and activation of HSCs. CONCLUSION:Our results suggest that during liver fibrosis,portal hypertension may induce the proliferation,migration and activation of HSCs through the up-regulation of mi R-9a-5p,which targets Sirt1.展开更多
AIM To evaluate the levels of mi R-192-5 p in non-alcoholic fatty liver disease(NAFLD) models and demonstrate the role of mi R-192-5 p in lipid accumulation. METHODS Thirty Sprague Dawley rats were randomly divided in...AIM To evaluate the levels of mi R-192-5 p in non-alcoholic fatty liver disease(NAFLD) models and demonstrate the role of mi R-192-5 p in lipid accumulation. METHODS Thirty Sprague Dawley rats were randomly divided into three groups, which were given a standard diet, a high-fat diet(HFD), and an HFD with injection of liraglutide. At the end of 16 weeks, hepatic mi R-192-5 p and stearoyl-Co A desaturase 1(SCD-1) levels were measured. Mi R-192-5 p mimic and inhibitor and SCD-1 si RNA were transfected into Huh7 cells exposed to palmitic acid(PA). Lipid accumulation was evaluated by oil red O staining and triglyceride assays. Direct interaction was validated by dual-luciferase reporter gene assays.RESULTS The HFD rats showed a 0.46-fold decrease and a 3.5-fold increase in hepatic mi R-192-5 p and SCD-1 protein levels compared with controls, respectively, which could be reversed after disease remission by liraglutide injection(P < 0.01). The Huh7 cells exposed to PA also showed down-regulation and up-regulation of mi R-192-5 p and SCD-1 protein levels, respectively(P < 0.01). Transfection with mi R-192-5 p mimic and inhibitor in Huh7 cells induced dramatic repression and promotion of SCD-1 protein levels, respectively(P < 0.01). Luciferase activity was suppressed and enhanced by mi R-192-5 p mimic and inhibitor, respectively, in wild-type SCD-1(P < 0.01) but not in mutant SCD-1. Mi R-192-5 p overexpression reduced lipid accumulation significantly in PA-treated Huh7 cells, and SCD-1 si RNA transfection abrogated the lipid deposition aggravated by mi R-192-5 p inhibitor(P < 0.01).CONCLUSION This study demonstrates that mi R-192-5 p has a negative regulatory role in lipid synthesis, which is mediated through its direct regulation of SCD-1.展开更多
基金Supported by National Natural Science Foundation of China,No.11272342/A0205
文摘AIM:To reveal the functions of micro RNAs(mi RNAs) with respect to hepatic stellate cells(HSCs) in response to portal hypertension.METHODS:Primary rat HSCs were exposed to static water pressure(10 mm Hg,1 h) and the pressureinduced mi RNA expression profile was detected by next-generation sequencing. Quantitative real-time polymerase chain reaction was used to verify the expression of mi RNAs. A potential target of Mi R-9a-5p was measured by a luciferase reporter assay and Western blot. CCK-8 assay and Transwell assay were used to detect the proliferation and migration of HSCs under pressure.RESULTS:According to the profile,the expression of mi R-9a-5p was further confirmed to be significantly increased after pressure overload in HSCs(3.70 ± 0.61 vs 0.97 ± 0.15,P = 0.0226),which resulted in the proliferation,migration and activation of HSCs. In vivo,the up-regulation of mi R-9a-5p(2.09 ± 0.91 vs 4.27 ± 1.74,P = 0.0025) and the down-regulation of Sirt1(2.41 ± 0.51 vs 1.13 ± 0.11,P = 0.0006) were observed in rat fibrotic liver with portal hypertension. Sirt1 was a potential target gene of mi R-9a-5p. Through restoringthe expression of Sirt1 in mi R-9a-5p transfected HSCs on pressure overload,we found that overexpression of Sirt1 could partially abrogate the mi R-9a-5p mediated suppression of the proliferation,migration and activation of HSCs. CONCLUSION:Our results suggest that during liver fibrosis,portal hypertension may induce the proliferation,migration and activation of HSCs through the up-regulation of mi R-9a-5p,which targets Sirt1.
基金Supported by National Key R&D Program of China No.2017YFC0908900National Key Basic Research Project,No.2012CB517501National Natural Science Foundation of China,No.81470840 and No.81600464
文摘AIM To evaluate the levels of mi R-192-5 p in non-alcoholic fatty liver disease(NAFLD) models and demonstrate the role of mi R-192-5 p in lipid accumulation. METHODS Thirty Sprague Dawley rats were randomly divided into three groups, which were given a standard diet, a high-fat diet(HFD), and an HFD with injection of liraglutide. At the end of 16 weeks, hepatic mi R-192-5 p and stearoyl-Co A desaturase 1(SCD-1) levels were measured. Mi R-192-5 p mimic and inhibitor and SCD-1 si RNA were transfected into Huh7 cells exposed to palmitic acid(PA). Lipid accumulation was evaluated by oil red O staining and triglyceride assays. Direct interaction was validated by dual-luciferase reporter gene assays.RESULTS The HFD rats showed a 0.46-fold decrease and a 3.5-fold increase in hepatic mi R-192-5 p and SCD-1 protein levels compared with controls, respectively, which could be reversed after disease remission by liraglutide injection(P < 0.01). The Huh7 cells exposed to PA also showed down-regulation and up-regulation of mi R-192-5 p and SCD-1 protein levels, respectively(P < 0.01). Transfection with mi R-192-5 p mimic and inhibitor in Huh7 cells induced dramatic repression and promotion of SCD-1 protein levels, respectively(P < 0.01). Luciferase activity was suppressed and enhanced by mi R-192-5 p mimic and inhibitor, respectively, in wild-type SCD-1(P < 0.01) but not in mutant SCD-1. Mi R-192-5 p overexpression reduced lipid accumulation significantly in PA-treated Huh7 cells, and SCD-1 si RNA transfection abrogated the lipid deposition aggravated by mi R-192-5 p inhibitor(P < 0.01).CONCLUSION This study demonstrates that mi R-192-5 p has a negative regulatory role in lipid synthesis, which is mediated through its direct regulation of SCD-1.
