LncRNA SNHG16通过靶向miR-421/MPC-1轴改善脓毒症诱导的肾炎和细胞凋亡  

LncRNA SNHG16 improving sepsis-induced nephritis and apoptosis by targeting miR-421/MPC-1 axis

作　　者：杨丹平 郭峻氚[1] 刘艳[1] 肖东[1] Yang Danping;Guo Junchuan;Liu Yan;Xiao Dong(Department of Critical Medicine,People's Hospital of Xinjiang Uygur Autonomous Region,Urumqi 830000,China)

机构地区：[1]新疆维吾尔自治区人民医院重症医学科,乌鲁木齐830000

出　　处：《广西医科大学学报》2023年第7期1107-1115,共9页Journal of Guangxi Medical University

基　　金：新疆维吾尔自治区自然科学基金项目(No.2021D01C156)。

摘　　要：目的:探讨长链非编码RNA(lncRNA)SNHG16对脓毒症诱导的肾细胞炎症和细胞凋亡的改善作用与机制。方法:应用内毒素脂多糖(LPS)在体外诱导人近端肾小管上皮细胞(HRPTEC)构建脓毒症样肾细胞模型,细胞分为对照组和LPS组。将LPS组的HRPTEC分为SNHG16过表达组(LPS+SNHG16组),过表达空载体组(LPS+vector组),SNHG16过表达联合miR-421拟似物处理组(LPS+SNHG16+miR-421 mimic组),SNHG16过表达联合线粒体丙酮酸载体-1(MPC-1)小干扰RNA(siRNA)沉默处理组(LPS+SNHG16+si-MPC-1组)。应用荧光素酶报告基因检测验证miR-421序列与SNHG16序列的结合情况以及miR-421与MPC-1 3’-UTR的结合情况。用实时荧光定量PCR(RT-qPCR)检测各组HRPTEC中SNHG16和miR-421表达水平,western blotting检测各组HRPTEC中cleaved-caspase 3、Bcl-2、MPC-1的表达水平,细胞计数试剂盒(CCK-8)法检测各组HRPTEC的增殖率,流式细胞术检测细胞凋亡率。酶联免疫吸附实验(ELISA)检测各组细胞培养上清中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)的水平。结果:与对照组比较,LPS组的细胞增殖率降低(P<0.05),细胞凋亡率增加(P<0.05),细胞中miR-421、cleaved-caspase 3和Bcl-2的表达增加(均P<0.05),而细胞中SNHG16和MPC-1的表达减少(均P<0.05),细胞培养上清中TNF-α、IL-1β、IL-6的表达增加(均P<0.05)。LPS处理联合SNHG16过表达增加了细胞增殖率(P<0.05),减少了细胞凋亡(P<0.05),抑制了细胞培养上清中TNF-α、IL-1β、IL-6的表达(均P<0.05)。而SNHG16组的上述作用均被miR-421-mimic部分逆转(P<0.05)。在LPS+SNHG16中加入si-MPC-1后,LPS+SNHG16组的上述作用均被si-MPC-1部分逆转(均P<0.05)。结论:LncRNA SNHG16通过靶向miR-421/MPC-1轴改善脓毒症诱导的肾细胞炎症和细胞凋亡。Objective:To investigate the effect and mechanism of long non-coding RNA(lncRNA)SNHG16 on sepsis-induced renal cell inflammation and apoptosis.Methods:Human renal proximal tubular epithelial cells(HRPTEC)was induced by lipopolysaccharide(LPS)in vitro to construct a sepsis-like renal cell model.The cells were divided into control group and LPS group.HRPTEC in LPS group were divided into SNHG16 overexpression group(LPS+SNHG16 group),overexpression empty vector group(LPS+vector group),SNHG16 overexpression combined with miR-421 mimic group(LPS+SNHG16+miR-421 mimic group)and SNHG16 overexpression combined with mitochondrial pyruvate carrier-1(MPC-1)small interfering RNA(siRNA)silencing group(LPS+SNHG16+si-MPC-1 group).Luciferase reporter gene detection was used to verify the binding of miR-421 sequence to SNHG16 sequence and the binding of miR-421 to MPC-13’-UTR.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression levels of SNHG16 and miR-421 in HRPTEC of each group,western blotting was used to detect the expression levels of cleaved-caspase 3,Bcl-2 and MPC-1 in HRPTEC of each group,cell counting kit-8(CCK-8)method was used to detect the proliferation rate of HRPTEC of each group and the apoptosis rate was detected by flow cytometry.The levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)in cell culture supernatant of each group were detected by enzyme-linked immunosorbent assay(ELISA).Results:Compared with the control group,the cell proliferation rate of LPS group decreased(P<0.05),the apoptosis rate increased(P<0.05),and the expressions of miR-421,cleaved-caspase 3 and Bcl-2 increased(all P<0.05).However,the expressions of SNHG16 and MPC-1 in cells decreased(all P<0.05),and the expressions of TNF-α,IL-1βand IL-6 in cell culture supernatant increased(all P<0.05).LPS treatment combined with SNHG16 overexpression increased the cell proliferation rate(P<0.05),decreased the apoptosis(P<0.05),and inhibited the expressions of TNF-�

关 键 词：长链非编码RNA SNHG16 微小RNA-421 线粒体丙酮酸载体1 脓毒症急性肾损伤 细胞凋亡 

分 类 号：R692[医药卫生—泌尿科学]