lncRNA PCAT-1通过调控HMGB1诱导的肝细胞焦亡在急性肝损伤中的作用机制研究  

The mechanism of lncRNA PCAT-1 regulating HMGB1 induced hepatocyte pyroptosis in acute liver injury

作　　者：周青青 吴捷 李节繁 ZHOU Qingqing;WU Jie;LI Jiefan(Department of Infectious,People’s Hospital of Pingyang,Wenzhou 325400,China)

机构地区：[1]平阳县人民医院感染科,浙江温州325400

出　　处：《健康研究》2025年第1期79-83,F0003,共6页Health Research

基　　金：温州市科技计划项目(Y20210862)。

摘　　要：目的探讨长链非编码RNA(lncRNA)前列腺癌相关转录因子1(PCAT-1)通过调控高迁移率族蛋白1(HMGB1)诱导的肝细胞焦亡在急性肝损伤中的作用机制。方法将L02肝细胞分成5组:正常对照组、模型组、si PCAT-1组、si NC组、si PCAT-1+anti-miR-129-5p组,后4组以10 mmol/L四氯化碳建立肝细胞急性损伤模型。24 h后检测培养上清中ALT、AST水平,Hoechst染色观察细胞形态;CCK-8检测细胞活力,RT-PCR检测lncRNA PCAT-1和miR-129-5p、HMGB1 mRNA表达水平,Western blot检测细胞焦亡相关蛋白NLRP3、Caspase-1表达水平。结果si lncRNA PCAT-1组的肝细胞活力为(62.71±5.48)%,低于其他4组;上清中ALT、AST水平分别为(3.45±0.73)U/L、(3.59±0.61)U/L,均高于其他4组;差异均有统计学意义(均P<0.001)。荧光显微镜下观:正常对照组肝细胞染色质正常,偶见蓝色碎片;模型组肝细胞染色质变小,蓝色或亮蓝色碎片增多;si lncRNA PCAT-1组肝细胞染色质变小,蓝色或亮蓝色碎片增多更为明显,损伤程度较模型组重;si NC组、si lncRNA PCAT-1+anti-miR-129-5p组细胞形态表现与模型组接近。各组lncRNA PCAT-1、miR-129-5p、HMGB1 mRNA的表达水平,差异均有统计学意义(均P<0.001);均为正常对照组最低(0.97±0.09、1.04±0.12、1.27±0.12),si lncRNA PCAT-1组最高(1.79±0.18、1.69±0.22、2.14±0.32)。结论lncRNA PCAT-1通过影响miR-129-5p表达来调控HMGB1诱导的肝细胞焦亡,继而诱发或加重急性肝损伤。Objective To investigate the mechanism by which long non-coding RNA(lncRNA)prostate cancer-associated transcript 1(PCAT-1)regulates high mobility group box 1(HMGB1)-induced hepatocyte pyroptosis in acute liver injury.Methods L02 hepatocytes were divided into five groups:normal control group(no special treatment),model group,si-PCAT-1 group,si-NC group,and si-PCAT-1+anti-miR-129-5p group.Acute liver injury was induced in the latter four groups using 10 mmol/L carbon tetrachloride(CCl4).After 24 hours,levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in the culture supernatant were measured,cell morphology was observed via Hoechst staining,cell viability was assessed using the CCK-8 assay,expression levels of lncRNA PCAT-1,miR-129-5p,and HMGB1 mRNA were determined by RT-PCR,and expression levels of pyroptosis-related proteins NLRP3 and Caspase-1 were evaluated by Western blot.Results Hepatocyte viability in the si-lncRNA PCAT-1 group was(62.71±5.48)%,significantly lower than in the other four groups.The supernatant levels of ALT and AST in the si-lncRNA PCAT-1 group were(3.45±0.73)U/L and(3.59±0.61)U/L,respectively,both higher than those in the other groups,with statistically significant differences(all P<0.001).Under fluorescence microscopy,chromatin in the normal control group appeared normal with occasional blue fragments;in the model group,chromatin was condensed with increased blue or bright blue fragments;in the si-lncRNA PCAT-1 group,chromatin condensation and blue/bright blue fragments were more pronounced,indicating greater damage compared to the model group;the si-NC and si-lncRNA PCAT-1+anti-miR-129-5p groups showed morphological features similar to the model group.The expression levels of lncRNA PCAT-1,miR-129-5p,and HMGB1 mRNA differed significantly across groups(all P<0.001),with the lowest levels observed in the normal control group(0.97±0.09,1.04±0.12,1.27±0.12)and the highest in the si-lncRNA PCAT-1 group(1.79±0.18,1.69±0.22,2.14±0.32).Conclusion lncRNA PCAT-1 regul

关 键 词：急性肝损伤 焦亡 长链非编码RNA 前列腺癌相关转录因子1 高迁移率族蛋白1 

分 类 号：R575[医药卫生—消化系统]