摘要
目的研究油酸对非酒精性脂肪肝细胞活性氧、周期及凋亡的影响。方法处理组用60μg/ml油酸处理小鼠正常肝细胞AML-1224、48、72h,对照组不作任何处理。检测细胞培养后上清甘油三酯、丙氨酸氨基转移酶、天冬氨酸氨基转移酶生化指标,油红O染色观察非酒精性脂肪肝细胞脂滴形成。EdU染色观察肝细胞增殖,利用流式细胞术检测肝细胞ROS、周期及凋亡。结果在24、48及72h时,处理组甘油三酯含量比对照组高(P<0.05)。天冬氨酸氨基转移酶和丙氨酸氨基转移酶24、48h结果变化比较,差异无统计学意义(P>0.05),在72h时升高(P<0.05)。油红O染色显示脂滴形成明显(P<0.05)。油酸处理72h后EdU染色显示进入S期细胞增多(P<0.05)。流式结果显示ROS比对照组升高(P=0.000),G 0/G 1期细胞降低(P<0.01),S期细胞数量增多(P<0.05),G 2/M期没有变化(P>0.05),同时肝细胞凋亡比对照组高(P=0.000),坏死比对照组高(P<0.01)。结论非酒精性脂肪肝细胞油酸的代谢产生大量ROS,可能使细胞阻滞在S期,促使了肝细胞的凋亡和坏死。
Objective To study the effect of oleic acid on cycle and apoptosis of nonalcoholic fatty liver hepatocytes.Methods The treatment group was treated with 60μg/ml oleic acid for 24,48 and 72h,while the control group was not treated.The biochemical indexes of triglyceride,alanine aminotransferase and aspartate aminotransferase were detected,and the formation of lipid droplets in nonalcoholic fatty liver cells was observed by oil red O staining.The proliferation of hepatocytes was observed by EdU staining,and ROS,the cell cycle and apoptosis were detected by flow cytometry.Results At 24,48 and 72h,the content of triglyceride in the treatment group was higher than that in the control group(P<0.05).Aspartate aminotransferase and alanine aminotransferase did not change at 24 and 48h(P>0.05),but increased at 72h(P<0.05).Oil red O staining showed the lipid droplets formation was obvious(P<0.05).After oleic acid treatment for 72h,EdU staining showed that the number of cells entering S phase increased(P<0.05).Flow cytometry results showed that ROS was higher than that in control group(P=0.000).G 0/G 1 phase cells decreased(P<0.01),and S phase cells increased(P<0.05),and G 2/M phase cells did not change(P>0.05).Meanwhile,hepatocytes apoptosis was higher than that in control group(P=0.000),and necrosis was also higher than that in control group(P<0.01).Conclusion In the nonalcoholic fatty liver cell model,the product ROS of oleic acid metabolism might induce cell cycle arrest at the S phase,and accelerate apoptosis and necrosis.
作者
李建华
侯静
黄登亮
田美媛
张耀刚
刘哲
马艳艳
Li Jianhua;Hou Jing;Huang Dengliang(The Central Laboratory,Qinghai University Affiliated Hospital,Qinghai 810000,China)
出处
《医学研究杂志》
2021年第11期80-85,共6页
Journal of Medical Research
基金
青海省科技计划项目(2019-ZJ-922)。
