沉默长链非编码RNA牛磺酸上调因子1对高糖诱导的小胶质细胞焦亡作用机制的研究  

Effect of silencing lncRNA TUG1 on pyroptosis induced by high glucose in microglia PAN

作　　者：潘友卓 郭文惠 雷皓月 鲁珣 张琦[3] Youzhuo;GUO Wenhui;LEI Haoyue(The First Clinical Medical College of Gansu University of Chinese Medicine,Lanzhou 730000,China)

机构地区：[1]甘肃中医药大学第一临床医学院,兰州730000 [2]宁夏医科大学临床学院 [3]甘肃省人民医院老年医学科

出　　处：《中国糖尿病杂志》2024年第10期764-769,共6页Chinese Journal of Diabetes

基　　金：国家自然科学基金(82160166、81960173);兰州市人才创新创业项目(2021-RC-136);兰州市卫生健康科技发展项(2021005)。

摘　　要：目的 探讨长链非编码RNA(lncRNA)牛磺酸上调因子1(TUG1)参与高糖(HG)诱导的小胶质(BV2)细胞焦亡的作用机制.方法 小鼠BV2细胞分为正常对照(NC)组、HG组、TUG1慢病毒基因沉默载体组(sh-TUG1)、慢病毒空载体组(sh-Con)、HG+sh-Con组、HG+sh-TUG1组.RT-qPCR检测TUG1 mRNA表达和转染效率,RT-qPCR和Western blot法检测核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、胱天蛋白酶-1(Caspase-1)、消皮素D(GSDMD)、IL-18、IL-1β mRNA和蛋白表达.结果 HG组TUG1 mRNA表达高于NC组(P<0.05).转染后sh-TUG1、sh-Con组出现大量绿色荧光,NC组细胞无绿色荧光,sh-TUG1 组TUG1 mRNA表达低于NC组(P<0.05),重组慢病毒成功感染BV2 细胞.HG、HG+sh-Con组NLRP3、Caspase-1、GSDMD、IL-18、IL-1β mRNA和蛋白表达高于NC组(P<0.05).HG+sh-TUG1 组NLRP3、Caspase-1、GSDMD、IL-18、IL-1β mRNA和蛋白表达低于HG、HG+sh-Con组(P<0.05).结论 TUG1 参与HG诱导的BV2 细胞焦亡,引起炎性反应,沉默TUG1可抑制此过程.Objective To investigate the mechanism of action of the long non‑coding RNA(lncRNA)taurine up‑regulating factor 1(TUG1)with high glucose(HG)‑induced cellular pyroptosisin microglial cell.Methods Mouse BV2 cells were cultured and divided into normal control(NC),HG,lentiviral empty vector(sh‑Con),TUG1 lentiviral gene silencing vector(sh‑TUG1),HG+sh‑Con and HG+sh‑TUG1 group.RT‑PCR was used to detect the expression and transfection efficiency of TUG1 mRNA.Nucleotide‑binding oligomerized structural domain‑like receptor protein 3(NLRP3),cysteoaspartate protease‑1(Caspase‑1),and ghrelin D(GSDMD),IL‑18,IL‑1βmRNA and protein expression were detected by RT‑PCR and Western blot.Results TUG1 mRNA expression was higher in HG group than in NC group(P<0.05).After transfection,a lot of green fluorescence appeared in sh‑TUG1 and sh‑Con group,while no green fluorescence was observed in NC group.The expression of TUG1 mRNA was lower in sh‑TUG1 group than in NC group(P<0.05).Accordingly,the recombinant lentivirus successfully infected BV2 cell.The expressions of the mRNA and protein of NLRP3,Caspase‑1,GSDMD,IL‑18 and L‑1βwere higher in HG,HG+sh‑Con groups than in NC group(P<0.05).The expressions of the mRNA and protein of NLRP3,Caspase‑1,GSDMD and IL‑18 and L‑1βwere lower in HG+shTUG1 group than in HG,HG+sh‑Con groups(P<0.05).Conclusions TUG1 is involved in high glucose induced pyroptosis in microglia and leads to inflammatory response. Silencing TUG1 can inhibit the pathological reaction.

关 键 词：小胶质细胞 长链非编码RNA牛磺酸上调因子1 细胞焦亡 消皮素D 

分 类 号：R587.2[医药卫生—内分泌] R749.2[医药卫生—内科学]