LncRNAHOTAIR靶向miR-17-5p/TXNIP对狼疮性肾炎进展的机制研究

Mechanism of LncRNAHOTAIR in promoting progression of lupus nephritis by targeting miR-17-5p/TXNIP

目的探讨长链非编码RNA同源盒基因转录反义RNA(LncRNAHOTAIR)靶向miR-17-5p/硫氧还蛋白相互作用蛋白(TXNIP)对狼疮性肾炎(LN)进展的机制。方法取60只MRL/lpr雌性小鼠,随机分为模型组、LncRNAHOTAIR敲低组、miR-17-5p agomir组、阴性对照组和LncRNAHOTAIR敲低+miR-17-5p antagomir组,每组12只,另取12只C57BL/6J雌性小鼠作为对照组,检测各组小鼠肾功能、免疫器官指数;以HE染色实验检测各组小鼠肾组织病理形态;以酶联免疫吸附测定(ELASA)法测定各组小鼠血清抗双链DNA(dsDNA)、免疫细胞因子免疫球蛋白G(IgG)、白细胞介素-6(IL-6)、单核细胞趋化蛋白1(MCP-1)及肿瘤坏死因子-α(TNF-α)水平;以实时荧光定量多聚核苷酸链式反应(qPCR)和免疫印迹法检测各组小鼠肾组织LncRNAHOTAIR、miR-17-5p及TXNIP表达。以双荧光素酶报告基因实验检测小鼠肾小球系膜细胞中LncRNAHOTAIR对miR-17-5p、miR-17-5p对TXNIP的靶向调节。结果与对照组比较,模型组小鼠肾组织发生严重病理损伤,胸腺指数、脾脏指数、miR-17-5p表达降低(P<0.05),尿蛋白浓度、血尿素氮(BUN)、血清抗dsDNA及IgG、IL-6、MCP-1、TNF-α、肾组织LncRNAHOTAIR及TXNIP表达升高(P<0.05)。与模型组比较,Ln⁃cRNAHOTAIR敲低组、miR-17-5p agomir组小鼠肾组织病理损伤减轻,胸腺指数、脾脏指数、miR-17-5p表达升高(P<0.05),尿蛋白浓度、BUN、血清抗dsDNA及IgG、IL-6、MCP-1、TNF-α、肾组织LncRNAHOTAIR及TXNIP表达降低(P<0.05)。下调miR-17-5p可减弱敲低LncRNAHOTAIR组对模型组小鼠各指标的作用。结论敲低LncRNAHOTAIR可通过上调miR-17-5p而降低TXNIP表达,进而减少LN小鼠免疫炎性因子产生,增强其免疫功能,抑制体内炎症发生发展,最终减轻小鼠肾组织损伤并改善其肾功能。

Objective To investigate the mechanism of long non-coding RNA metastasis homeobox transcript antisense RNA(LncRNA HOTAIR)in promoting the progression of lupus nephritis(LN)by targeting the miR-17-5p/thioredoxin interacting protein(TXNIP)axis.Methods Sixty MRL/lpr female mice were randomly separated into the model group,LncRNAHOTAIR knockdown group,miR-17-5p agamir,negative control group,and LncRNAHOTAIR knockdown+miR-17-5p agamir group,with 12 mice in each group,and another 12 C57BL/6J female mice were collected as the control group.After grouping and intervention treatment,renal function,and immune organ index of mice in each group were detected;HE staining experiment was applied to detect the pathological morphology of kidney tissue of mice in each group;enzyme-linked immunosorbent assay(ELASA)method was applied to measure the serum levels of anti double stranded DNA(dsDNA)and immune cytokines such as immunoglobulin G(IgG),interleukin-6(IL-6),monocyte chemotactic protein-1(MCP-1),and tumor necrosis factor-α(TNF-α)of mice in each group;quantitative real-time PCRpolymerase chain reaction(qPCR)and immunoblotting experiments were applied to detect the expression of LncRNAHOTAIR,miR-17-5p,and TXNIP in the kidney tissue of mice in each group.Dual luciferase reporter gene experiment was applied to detect the targeted regulation of LncRNAHOTAIR on miR-17-5p,and miR-17-5p on TXNIP in mouse mesangial cells.Results Compared with the control group,the kidney tissue of mice in the model group experienced severe pathological damage,the thymus index,spleen index,and miR-17-5p expression decreased(P<0.05),and the urinary protein concentration,blood urea nitrogen(BUN)level,serum anti-dsDNA and IgG,IL-6,MCP-1,TNF-αlevels,and renal tissue LncRNAHOTAIR and TXNIP expression increased(P<0.05).Compared with the model group,the pathological damage to the kidney tissue of mice in the LncRNAHOTAIR knockdown group and miR-17-5p agomir group was reduced,the thymus index,spleen index,and miR-17-5p expression increased(P<0.05),and the urinary protein concentration,BUN level,serum anti-dsDNA and IgG,IL-6,MCP-1,TNF-αlevels,and renal tissue LncRNAHOTAIR and TXNIP expression decreased(P<0.05).Downregulating miR-17-5p could weaken the effects of knocking down LncRNAHOTAIR on various indicators in the model group mice.Conclusions Knocking down LncRNA HOTAIR can reduce TXNIP expression by up-regulating miR 17-5p,thereby reducing the production of immune inflammatory factors in LN mice,enhancing their immune function,inhibiting the occurrence and development of inflammation in vivo,and ultimately reducing renal tissue damage in mice and improving their renal function.