熊果酸对瘦素诱导的肝星状细胞JAKSTAT3活化-第三军医大学学报.DOC

熊果酸对瘦素诱导的肝星状细胞JAK2-STAT3活化及活性氧产生的影响 刘戈云 何文华 李弼民 李博 张新华 陈江 张焜和 朱萱 【摘要】 目的JAK2-STAT3信号通路活化、活性氧（ROS）产生及TIMP-1、MMP-1 mRNA。方法瘦素HSC-T6细胞，干预组在瘦素作用前分别加入熊果酸、N-乙酰-L半胱氨酸 (NAC)、氯化二亚苯基碘嗡（DPI）AG490干预，正常对照组不加任何药物。各组处理不同时间后，采用流式细胞细胞内采用免疫细胞化学检测采用RT-PCR检测结果. 免疫细胞化学瘦素-T6细胞2h后p-JAK2及细胞核内p-STAT3蛋白表达与正常对照组P0.01）熊果酸p-JAK2及p-STAT3蛋白表达明显低于瘦素刺激组（85.50±3.78比96.67±3.01，26.50±3.66比86.67±10.87，均P0.01）。NAC、DPI 、AG490干预p-JAK2及 p-STAT3蛋白表达也低于瘦素刺激组P0.05或P0.01） 2. 流式细胞术检测显示，熊果酸NAC、DPI和AG490干预的瘦素刺激组±5.58、55.56±6.43、36.54±3.21、48.12±6.63比90.86±3.86，均P0.01)，而且熊果酸干预组的NAC干预组P0.01)。3.熊果酸干预12h h后与瘦素刺激组mRNA表达0.28±0.03比0.37±0.01，0.29±0.04比0.41±0.03，均P0.01），MMP-1mRNA的表达明显（0.33±0.00.26±0.02，0.37±0.04比0.22±0.04，均P0.01）结论JAK2-STAT3信号通路活化及细胞内ROS的产生，下调TIMP-1mRNA的表达上调MMP-1mRNA表达 【关键词熊果酸；肝星状细胞The effect of ursolic acid on leptin-indused JAK2-STAT3 activation and Reactive oxygen species generation in Hepatic stellate cell LIU Ge-yun, HE Wen-hua, LI Bi-min, LI Bo, ZHANG Xin-hua, Chen Jiang, ZHANG Kun-he, ZHU Xuan. (Department of Gastroenterology, First Affliated Hospital of Nanchang University, Nanchang，330006,China). Corresponding author： ZHU Xuan， Email: jyyfyzx@163.com ;0791-8692505 【Abstract】　Objective To observe the effects of Ursolic acid(UA) on JAK2-STAT3 activation, ROS production and the gene expression of TIMP-1 and MMP-1 in Hepatic stellate cell(HSC) induced leptin, and explore the molecular mechanisms of UA on treatment of liver fibrosis. Methods　HSC-T6 cells or not with leptin(100ng/ml); or pretreatment with inhibitors, such as UA（50uM, NAC（10mM）, DPI（20uM）, AG490 for 30min,then with leptin. The phosphorylation of Janus kinase2(JAK2) and the nuclear translocation of signal transducers and activators of transcription 3(STAT3) protein expression were evaluated by immunocytochemical staining assay; Intracellular ROS levels were measured by using flow cytometry to detect DCF fluorescence; The mRNA levels of TIMP-1 and MMP-1 were evaluated by RT-PCR.Results UA decrease the p-JAK2 and p-STAT3 protein expre