摘要
目的研究体外药物性肝损伤肝细胞模型miR-122干预后肝损伤指标、肝细胞增殖以及IL-6水平的变化,探索miR-122在药物性肝损伤肝细胞的可能作用及机制。方法将肝细胞分为Ctrl组、APAP组、APAP+mimic-NC组、APAP+miR-122 mimic组、APAP+inhibitor-NC组、APAP+miR-122 inhibitor组6组,分别取各组细胞上清液,通过PCR法、免疫组化法等分别检测miR-122、AST、ALT、IL-6表达水平及细胞增殖水平。结果(1)miR-122水平比较:APAP+miR-122 mimic组miR-122表达水平最高,APAP+miR-122 inhibitor组miR-122表达水平最低,提示细胞转染成功,APAP组miR-122表达水平与APAP+mimic-NC组、APAP+inhibitor-NC组比较,差异无统计学意义(P>0.05)。(2)细胞增殖水平比较:Ctrl组细胞增殖水平最高,APAP+miR-122 inhibitor组细胞增殖水平最低,APAP组与APAP+mimic-NC组、APAP+inhibitor-NC组细胞增殖水平差异无统计学意义(P>0.05),与APAP+miR-122 mimic组细胞增殖水平差异有统计学意义(P<0.05)。(3)肝损伤指标比较:Ctrl组AST、ALT水平最低;APAP+miR-122 inhibitor组AST、ALT水平最高;APAP组与Ctrl组、APAP+miR-122 mimic组、APAP+miR-122 inhibitor组AST、ALT水平之间差异有统计学意义(P<0.05);与APAP+mimic-NC组、APAP+inhibitor-NC组AST、ALT水平之间差异无统计学意义(P>0.05)。(4)IL-6表达水平比较IHC结果显示:Ctrl组IL-6表达水平最低,APAP+miR-122 inhibitor组IL-6表达水平最高;APAP组与APAP+miR-122 mimic组、APAP+miR-122 inhibitor组之间IL-6表达水平差异有统计学意义(P<0.05),与APAP+mimic-NC组、APAP+inhibitor-NC组之间IL-6表达水平差异无统计学意义(P>0.05)。结论miR-122可能通过调控IL-6实现对DILI肝细胞的保护作用。
Objective To explore the changes of liver injury indexes,hepatocyte proliferation and IL-6 level after miR-122 intervention in hepatocyte model of drug-induced liver injury in vitro,then in order to explore the possible role and mechanism of miR-122 on the hepatocytes in drug-induced liver injury.Methods The hepatocytes were divided into 6 groups:Ctrl group,APAP group,APAP+mimic-NC group,APAP+miR-122 mimic group,APAP+inhibitor-NC group and APAP+miR-122 inhibitor group.The expression levels of miR-122,AST,ALT,IL-6 and cell proliferation were detected by PCR and immunohistochemical methods.Results(1)Comparison of miR-122 level:the expression level of miR-122 was the highest in APAP+miR-122 mimic group,and the lowest in APAP+miR-122 inhibitor group,indicating successful transfection of cells.The expression level of miR-122 in APAP group was higher than that in Ctrl group(P<0.05),there was no significant difference between APAP+mimic-NC group and APAP+inhibitor-NC group(P>0.05).(2)Comparison of cell proliferation:cell proliferation in Ctrl group was the highest,and in APAP+miR-122 inhibitor group was the lowest.There was no significant difference in cell proliferation between APAP group and APAP+mimic-NC group and APAP+inhibitor-NC group.The cell proliferation level was significantly different from that of APAP+miR-122 mimic group(P<0.05).(3)Comparison of liver injury indexes:the levels of AST and ALT in the Ctrl group were the lowest.The AST and ALT levels of APAP+miR-122 inhibitor group were the highest;APAP group with Ctrl group and APAP+miR-122 mimic group.There were significant differences in AST and ALT levels between APAP+miR-122 mimic group and APAP+miR-122 inhibitor groups(P<0.05).With APAP+mimic-NC group,APAP+inhibitor-NC group.There was no significant difference between AST and ALT levels(P>0.05).(4)Immunohistochemical results:the IHC results showed that the expression of IL-6 was the lowest in Ctrl group,and the highest in APAP+miR-122 inhibitor group.The expression level of IL-6 was significant between A
作者
李娟娟
艾克鹏
杨佳
古巧燕
张海斌
LI Juanjuan;AI Kepeng;YANG Jia;GU Qiaoyan;ZHANG Haibin(Department of Gastroenterology,Yan'an University Affiliated Hospital,Yan'an 716000;Department of Gastroenterology,Shanghai East Hospital,China)
出处
《胃肠病学和肝病学杂志》
CAS
2024年第11期1482-1486,共5页
Chinese Journal of Gastroenterology and Hepatology
基金
2020年度陕西省留学人员科技活动择优资助项目(2020014)
陕西省重点研发计划一般项目—社会发展领域(2017SF-157)。
