circMTO1靶向miR-384减轻氯氮平致大鼠原代肝细胞损伤的机制研究  

Mechanism of circMTO1 in alleviating clozapine-induced rat primary hepatocytes injury by targeting miR-384

作　　者：张卫华[1] 张子雯[1] 张鹏[1] ZHANG Wei-hua;ZHANG Zi-wen;ZHANG Peng(School of Medicine,Yichun University,Yichun Jiangxi 336000,China)

机构地区：[1]江西省宜春学院医学院,江西宜春336000

出　　处：《毒理学杂志》2023年第1期31-36,共6页Journal of Toxicology

基　　金：2020年江西省教育厅科学技术研究项目(201625)。

摘　　要：目的 探讨环状RNA(circRNA)circMTO1对氯氮平致大鼠原代肝细胞损伤的影响及分子机制。方法 将大鼠原代肝细胞随机分为对照组、氯氮平组、氯氮平+pcDNA3.1组、氯氮平+pcDNA3.1-circMTO1组、氯氮平+miR-NC组、氯氮平+miR-384组和氯氮平+pcDNA3.1-circMTO1+miR-384组。采用细胞计数试剂盒-8(CCK-8)实验检测细胞存活率,实时荧光聚合酶链式反应(qRT-PCR)检测circMTO1和miR-384表达水平,流式细胞术检测细胞凋亡率,TUNEL染色检测细胞凋亡指数,酶联免疫吸附试验(ELISA)检测细胞上清液中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平,双荧光素酶报告实验验证circMTO1与miR-384的靶向关系。结果 对照组、氯氮平组、氯氮平+pcDNA3.1组、氯氮平+pcDNA3.1-circMTO1组、氯氮平+miR-NC组、氯氮平+miR-384组和氯氮平+pcDNA3.1-circMTO1+miR-384组大鼠原代肝细胞存活率依次为100.00%±5.37%、53.09%±8.71%、58.71%±9.26%、80.25%±12.71%、56.07%±8.02%、39.01%±5.64%、61.24%±8.35%,凋亡率依次为5.53%±0.93%、37.26%±6.20%、41.18%±5.60%、19.87%±3.03%、38.96%±5.02%、53.24%±6.01%和32.69%±4.17%。与对照组比较,氯氮平组大鼠原代肝细胞存活率和circMTO1表达水平明显降低,miR-384表达水平、凋亡率、凋亡指数、ALT和AST水平明显增加(P<0.05)。与氯氮平组比较,氯氮平+pcDNA3.1-circMTO1组大鼠原代肝细胞存活率和circMTO1表达水平明显增加,miR-384表达水平、凋亡率、凋亡指数、ALT和AST水平明显降低(P<0.05)。与氯氮平组比较,氯氮平+miR-384组大鼠原代肝细胞存活率明显降低,miR-384表达水平、凋亡率、凋亡指数、ALT和AST水平明显增加(P<0.05)。与氯氮平+pcDNA3.1-circMTO1组比较,氯氮平+pcDNA3.1-circMTO1+miR-384组大鼠原代肝细胞存活率明显降低,miR-384表达水平、凋亡率、凋亡指数、ALT和AST水平明显增加(P<0.05)。与氯氮平+miR-384组比较,氯氮平+pcDNA3.1-circMTO1+miR-384组大鼠原代肝细胞circMTO1表达�Objective To investigate the effect of Circulating RNA(circRNA) circMTO1 on clozapine-induced rat primary hepatocytes injury and its molecular mechanism. Methods Rat primary hepatocytes were randomly divided into control group, clozapine group, clozapine+pcDNA3.1 group, clozapine+pcDNA3.1-circMTO1 group, clozapine+miR-NC group, clozapine+miR-384 group and clozapine+pcDNA3.1-circMTO1+miR-384 group. Cell counting kit 8(CCK-8) assay was used to detect cell survival rate. Real-time fluorescence polymerase chain reaction(QRT-PCR) was used to detect the expression levels of circMTO1 and miR-384. Flow cytometry was used to detect cell apoptosis rate. TUNEL staining was used to detect cell apoptosis index. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) in cell supernatant. Double luciferase reporting assay was used to verify the targeting relationship between circMTO1 and miR-384. Results The survival rate of primary hepatocytes in control group, clozapine group, clozapine+pcDNA3.1 group, clozapine+pcDNA3.1-circMTO1 group, clozapine+miR-NC group, clozapine+miR-384 group and clozapine+pcDNA3.1-circMTO1+miR-384 group was 100.00%±5.37 %, 53.09%±8.71 %, 58.71%±9.26%, 80.25%±12.71%, 56.07%±8.02%, 39.01%±5.64% and 61.24%±8.35%, respectively. The apoptosis rate was 5.53%±0.93%, 37.26%±6.20%, 41.18%±5.60%, 19.87%±3.03%, 38.96%±5.02%, 53.24%±6.01% and 32.69%±4.17%, respectively. Compared with the control group, the survival rate and circMTO1 expression level of rat primary hepatocytes in clozapine group were significantly decreased, while miR-384 expression level, apoptosis rate, apoptosis index, ALT and AST levels were significantly increased(P<0.05). Compared with the clozapine group, the survival rate and circMTO1 expression level of rat primary hepatocytes in clozapine+pcDNA3.1-circMTO1 group were significantly increased, while miR-384 expression level, apoptosis rate, apoptosis index, ALT and AST levels were significant

关 键 词：circMTO1 miR-384 氯氮平 大鼠原代肝细胞 凋亡 

分 类 号：R114[医药卫生—卫生毒理学] R99[医药卫生—公共卫生与预防医学]