1,25(OH)_2D_3通过调节miR-146a水平抑制大鼠肝纤维化的体内和体外观察  

1,25(OH)_2D_3 inhibits hepatic fibrosis by regulating miR-146a levels in vitro and in vivo

作　　者：周丽云 李校天[2] 李丽[3] 杨俊超[3] Zhou Liyun;Li Xiaotian;Li Li(Department of Gastroenterology,Affiliated Hospital,Hebei University of Engineering,Handan 056002,Hebei Province,China)

机构地区：[1]浙江省嘉兴市第一医院肝病研究所,314000 [2]河北工程大学附属医院消化内科 [3]河北工程大学附属医院血液内科

出　　处：《实用肝脏病杂志》2019年第3期333-336,共4页Journal of Practical Hepatology

基　　金：河北省卫生与计划生育委员会重点科技研究计划项目(编号20150490);河北省研究生创新资助项目(编号171290080015)

摘　　要：目的体内体外观察1,25(OH)_2D_3通过调节mi R-146a水平抑制大鼠肝纤维化的作用机制。方法建立CCl_4诱导的大鼠肝纤维化模型,体外转染肝星状细胞(HSCs)mi R-146a模拟剂/抑制剂,观察1,25(OH)_2D_3处理对动物肝组织变化和HSC增殖和凋亡的影响。采用qPCR法检测肝组织mi R-146a水平,采用CCK8法检测细胞增殖,使用流式细胞术检测HSC凋亡。结果在干预8 w末,1,25(OH)_2D_3干预组大鼠肝纤维化程度明显减轻;1,25(OH)_2D_3干预组大鼠肝组织mi R-146a水平为(0.70±0.03),显著高于橄榄油组【(0.33±0.17,P<0.05)】;1,25(OH)_2D_3组细胞增殖率为58.8%,较DMSO组下降了15.9%,转染mi R-146a模拟剂组大鼠HSC增殖率为46.5%,较对照组下降了53.3%,转染mi R-146a抑制剂组HSC增殖率为132.8%,较对照物组升高了32.8%(P<0.05),1,25(OH)_2D_3干预组细胞凋亡率为12.6%,较DMSO组增加了5.2%,转染mi R-146a模拟剂组细胞凋亡率为16.8%,较对照组细胞凋亡率增加了8.2%,转染mi R-146a抑制剂组细胞凋亡率为6.3%,较对照组细胞凋亡率减少了2.2%(P<0.05),提示1,25(OH)_2D_3具有抑制HSC增殖、促进凋亡作用。结论 1,25(OH)_2D_3可能通过调节mi R-146a水平抑制HSC活化和抑制大鼠肝纤维化。Objective To observe the inhibitory effect of 1,25(OH)2D3 on liver fibrosis in hepatic stellate cells (HSCs) and in rat model. Method The CCl4-induced liver fibrosis was made in rats,and the activation of HSCs were induced by 10 pmmol/L TGF-β1 incubation. The miR-146a mimic/inhibitor were transfected in HSCs and 1,25(OH)2D3 intervention were paralleled. The miR-146a levels in liver tissues were detected by qPCR,the proliferation of cells was measured by CCK8 and the apoptosis was detected by flow cytometry. Results At the end of 8 weeks,the hepatic fibrosis in 1,25(OH)2D3 intervention group was significantly milder than that in the control group;the miR-146a level in 1,25(OH)2D3 intervention group was significantly higher than in oil control group[(0.70±0.03) vs.(0.33±0.17),P<0.05];the proliferation rate in1,25(OH)2D3 group was 58.8%,15.91% of lower than in DMSO group,in miR-146a mimic transfected group was 46.5%,53.3% of lower than in control,in miR-146a inhibitor group was132.8%,32.8% of higher than in control(P<0.05);the apoptosis rate of HSCs in1,25(OH)2D3 group was 12.6%,5.2% of higher than in DMSO,in miR-146a mimic transfected group was16.8%,8.2% of higher than in control,and in miR-146a inhibitor transfected group was 6.3%,2.2% of lower than in control (P<0.05),indicating the inhibition of 1,25(OH)2D3 on proliferation and promotion on apoptosis of HSCs. Conclusion 1,25(OH)2D3 might modulate miR-146a levels to inhibit liver fibrosis.

关 键 词：肝纤维化 肝星状细胞 1 25(OH)2D3 MIR-146A 大鼠 

分 类 号：R575.2[医药卫生—消化系统]