LncRNA CASC7/miR-21/Wnt2b轴介导糖尿病肾病小鼠肾纤维化的机制研究  

Mechanism of LncRNA CASC7/miR-21/Wnt2b axis mediating renal fibrosis in diabetic nephropathy mice

作　　者：连欢[1] 魏小宝 张娟[1] LIAN Huan;WEI Xiaobao;ZHANG Juan(Department of Nephrology,Zhongnan Hospital of Wuhan University,Wuhan,Hubei,China,430071)

机构地区：[1]武汉大学中南医院肾内科,湖北武汉430071

出　　处：《分子诊断与治疗杂志》2023年第1期60-64,共5页Journal of Molecular Diagnostics and Therapy

基　　金：湖北省自然科学基金面上项目(一般面上)(2020CFB634)。

摘　　要：目的 探究miR-21靶向调控肾纤维化的发生发展机制,为临床糖尿病肾病(DN)的治疗提供新的思路。方法 24只C57BLKS/J-Leprdb/db小鼠随机分为实验1组、实验2组与对照组,每组8只。实验1组进行单侧肾切除术,术后2月,实验1组和实验2组小鼠采用腹腔注射链脲佐菌素(STZ)的方法建立糖尿病肾病动物模型。HE染色法观察肾组织病理学改变,Masson染色法观察肾组织纤维化状况,qRT-PCR法检测各组肾组织中LncRNA CASC7、miR-21、Wnt2b、SMAD7表达水平,western blot法检测Wnt2b和SMAD7的蛋白表达水平,荧光素酶报告系统鉴定LncRNA CASC7和miR-21靶向关系。Pearson相关性分析肾组织中LncRNA CASC7和miR-21的表达水平与糖尿病肾病肾纤维化之间的关系。结果染色试验中观察到实验1组与实验2组小鼠肾小球肥大,系膜细胞及系膜基质增生,肾小管上皮细胞空泡变性。肾组织存在炎性细胞浸润、肾小管萎缩与间质纤维化。肾组织纤维化评分:实验1组(2.87±0.66)分>实验2组(2.17±0.62)分>对照组(0.57±0.14)分,差异有统计学意义(P>0.05);LncRNA CASC7、miR-21、Wnt2b、SMAD7表达水平:实验1组>实验2组>对照组,差异有统计学意义(P<0.05);与对照组相比,实验1组和实验2组Wnt2b和SMAD7蛋白表达均升高,且实验1组高于实验2组,差异有统计学意义(P<0.05);荧光素酶报告表明LncRNA CASC7可与miR-21靶向结合。Pearson相关性分析结果显示,肾组织中LncRNA CASC7和miR-21的表达水平与糖尿病肾病肾纤维化呈正相关(r=0.412、0.371,P均<0.05)。结论 LncRNA CASC7可通过调控下游miR-21改变Wnt2b、SMAD7的表达,从而参与调控Wnt2b、SMAD通路,并介导DN肾纤维化的发展。Objective To explore the mechanism of miR-21 targeting to regulate the occurrence and development of renal fibrosis,and to provide new ideas for clinical treatment of diabetic nephropathy(DN). Methods Twenty-four C57BLKS/J-Leprdb/db mice were randomly divided into experimental group 1,experimental group 2 and the control group,with 8 mice in each group. The experimental group 1 underwent unilateral nephrectomy,and 2 months after the operation,the mice in the experimental group 1 and the experimental group 2 were injected with streptozotocin(STZ)intraperitoneally to establish an animal model of diabetic nephropathy. HE staining was used to observe pathological changes of renal tissue,and Masson staining was used to observe renal fibrosis. qRT-PCR method was used to detect the expression levels of LncRNA CASC7,miR-21,Wnt2b and SMAD7 in the kidney tissue of each group. Western blot abalysis was used to detect the protein expression levels of Wnt2b and SMAD7. Identification of the targeting relationship between LncRNA CASC7 and miR-21 by a nelucerase reporter system was performed. Pearson correlation analysis was used to evaluate the relationship between the expression levels of LncRNA CASC7 and miR-21 in renal tissue and renal fibrosis in diabetic nephropathy. Results In the staining test,glomerular hypertrophy,mesangial cells and mesangial matrix hyperplasia,and vacuolar degeneration of renal tubular epithelial cells were observed in mice of group 1 and group 2. Inflammatory cell infiltration,tubular atrophy and interstitial fibrosis were found in the kidney. Kidney tissue fibrosis score:experimental group 1(2.87±0.66)> experimental group 2(2.17 ± 0.62)> control group(0.57 ± 0.14)(P<0.05). The expression levels of LncRNA CASC7,miR-21,Wnt2b,and SMAD7:experimental group 1 > experimental group 2 > control group,the difference was statistically significant(P<0.05). Compared with the control group,the protein expressions of Wnt2b and SMAD7 in the experimental group 1 and the experimental group 2 were increased,and th

关 键 词：糖尿病肾病 肾纤维化 LncRNA CASC7 MIR-21 Wnt2b 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]