PI3K抑制剂对四氯化碳刺激的肝星状细胞增殖和Ⅰ型胶原表达的影响  

Effects of phosphatidyl inositol 3-kinase inhibitors on proliferation and type Ⅰ collagen expression of hepatic stellate cell stimulated by carbon tetrachloride in vitro

作　　者：李娜[1] 祝聪聪[1] 肖永红[1] 张文丽[2] 刘亦民[2] 

机构地区：[1]河北联合大学公共卫生学院,河北唐山063000 [2]河北联合大学医学实验研究中心,河北唐山063000

出　　处：《西安交通大学学报（医学版）》2013年第2期164-167,共4页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：河北省自然科学基金资助项目(No.C2010001750)~~

摘　　要：目的探讨PI3K抑制剂对四氯化碳(CCl4)刺激的肝星状细胞(HSC)增殖和Ⅰ型胶原表达的影响,为肝纤维化的治疗提供理论基础。方法将液氮保存下的HSC株,于37℃,50mL/L CO2孵育箱中复苏传代培养,同步化后将细胞分为5组:空白对照组、CCl4组、CCl4+1μmol/L PI103组、CCl4+2μmol/L PI103组、CCl4+4μmol/L PI103组,分别培养48h。用透射电子显微镜初步观察了细胞形态改变;MTT比色法测定细胞增殖情况;免疫细胞化学测定细胞的Ⅰ型胶原表达情况。结果 CCl4组与空白对照组比较,细胞生长旺盛,细胞数目增多,Ⅰ型胶原表达增多;透射电子显微镜下可见CCl4+4μmol/L PI103组比CCl4组凋亡细胞显著增多,细胞数目减少;MTT和免疫细胞化学实验结果显示,与CCl4组比较,CCl4+1μmol/L PI103组、CCl4+2μmol/L PI103组、CCl4+4μmol/L PI103组细胞的增殖和Ⅰ型胶原的表达均减少,随着PI103剂量加大,细胞的增殖和Ⅰ型胶原的表达均减少,但CCl4+4μmol/L PI103组和CCl4+2μmol/L PI103组比较,细胞的增殖无明显变化。结论 PI3K抑制剂PI103可抑制活化的HSC的增殖,促进其凋亡以及减少其Ⅰ型胶原的表达。Objective To discuss the effects of phosphatidylinositol 3-kinase （PI3K） inhibitors on proliferation and type I collagen expression of hepatic stellate cell （HSC） stimulated by carbon tetrachloride（CCl ） in vitro and to lay theoretical basis for the treatment of liver fibrosis. Methods HSC lines were incubated in 50 mL/L CO2 incubation box at 37 ℃ for passage culture. Then they were divided into five groups： the control group, CC14 group, CCI ＋1 μmol/L PI103 group, CCI ＋2mol/L PII03 group and CCl4 ＋4 mol/L PI103 group, and cultured for 48 hours separately. Morphological changes of the cells were observed under the transmission electron microscope, cell proliferation was examined by MTT, and type I collagen expression was determined by immunocytochemistry. Restllts Compared with those in the control group, the cells grew well, cell number increased, and type I collagen expression increased in CCI group. Transmission electron microscopy showed that the apoptotic cells increased while the number of cells decreased in CC14 ＋ 4 μmol/L PI103 group compared with those in CC14 group. MTT and immunocytochemistry experiments results showed that cell proliferation and the expression of type I collagen in CC14 ＋1 μmol/L PI103 group, CC14 ＋2/μmol/L PI103 group and CCh ＋4/μmol/L PI103 groups all decreased when compared with CCL group. As the dosage of PI103 increased, both cell proliferation and the expression of type I collagen decreased, but there were no significant changes in cell proliferation in CCL ＋ 4 μmol/L PI103 group compared with CC14 ＋ 2 μmol/L PI103 group. Oonclusion PI3K inhibitors could inhibit cell proliferation, promote apoptosis and reduce type I collagen expression in activated HSCs.

关 键 词：PI3K抑制剂 肝星状细胞(HSC) 增殖 Ⅰ型胶原 肝纤维化 PI3K/AKT信号通路 

分 类 号：R181.2[医药卫生—流行病学]