PI3K抑制剂PI103对四氯化碳刺激的肝星状细胞凋亡和细胞内钙离子浓度的影响  

Effect of phosphatidyl inositol 3-kinase inhibitors PI103 on apoptosis and calcium ion concentration in carbon tetrachloride stimulated hepatic stellate cell

作　　者：李娜[1] 祝聪聪[1] 肖永红[1] 

机构地区：[1]河北联合大学公共卫生学院,河北唐山063000

出　　处：《工业卫生与职业病》2013年第5期272-275,共4页Industrial Health and Occupational Diseases

基　　金：河北省自然科学基金(C2010001750)

摘　　要：目的探讨PI3K抑制剂PI103对四氯化碳(CCl4)刺激的肝星状细胞(HSC)凋亡率和细胞内游离钙离子([Ca2+]i)浓度的影响,为肝纤维化的治疗提供理论基础。方法将HSC株于37℃、5%CO2孵育箱中复苏传代培养,同步化后将细胞分为3组:空白对照组、CCl4组、CCl4+PI3K抑制剂组。用锚定蛋白V异硫氰酸荧光素/碘化丙啶(Annexin V FITC/PI)双标记染色,通过流式细胞仪观察各组细胞凋亡情况;以钙离子荧光探针(Fluo-3AM)为[Ca2+]i荧光指示剂负载细胞,用激光扫描共聚焦显微镜观察[Ca2+]i变化情况。结果 CCl4作用后,HSC凋亡率的变化不明显(P>0.05),但[Ca2+]i荧光强度明显增强(P<0.05)。PI3K抑制剂作用于经CCl4刺激的HSC后,细胞凋亡率增加(P<0.05),[Ca2+]i荧光强度减弱(P<0.05)。结论 PI3K抑制剂PI103可增加HSC的凋亡率,降低[Ca2+]i浓度。Objective To discuss the effect of phosphatidyl inositol 3-kinase（PI3K）inhibitors PI103 on apoptosis and intracellular calcium ion [Ca^2＋] 3i concentration of carbon tetrachloride （CCl4）stimulated hepatic stellate cell（ HSC） in vitro, and provide the theoretical basis for treatment of liver fibrosis. Methods HSC lines were incubated in 5% CO2 incubation box at 37℃, which were afterward divided into three groups： the control group, CCl4 group and CCl4 q- PI3K inhibitors group. Apoptosis situation in each group was observed through the flow cytometric after Annexin V FITC/PI double marking dyeing. The change of [Ca^2＋] 3i was observed by the confocal laser scanning microscopy with Fluo-3 AM for [Ca^2＋]i fluorescence indicator load cells. Results After treated with CCl4, the apoptosis of HSC was not obvious （P〉0.05）, while the fluorescence intensity of [Ca^2＋]i enhanced obviously（P〈0.05）. After PI3K inhibitors were added into CCl4 stimulated HSC, the apoptosis rate increased （P〈0.05）, and the fluorescence intensity of [Ca^2＋]i decreased （P〈 0.05）. Conclusions PI3K inhibitors PI103 may increase the apoptosis and decrease the [Ca^2＋]i concentration in HSC.

关 键 词：PI3K抑制剂 肝星状细胞(HSC) 凋亡 钙离子 

分 类 号：R181.2[医药卫生—流行病学]