LncRNA SNHG14调控miR-101-3p/SGK1轴对高糖诱导的肾小球系膜细胞增殖及纤维化的影响OA
The Effect of LncRNA SNHG14 on High Glucose Induced Proliferation and Fibrosis of Glomerular Mesangial Cells by Regulating the miR-101-3p/SGK1 Axis
目的:探究LncRNA SNHG14 调控miR-101-3p/SGK1 轴对高糖(HG)诱导的肾小球系膜细胞(GMC)增殖及纤维化的影响.方法:收集糖尿病肾病(DN)患者(DN组)和健康体检者(NC 组)血清,RT-qPCR检测血清中SNHG14、miR-101-3p、SGK1 mRNA表达.培养人GMC(HGMC),并分为NG组、HG组、HG+SNHG14 敲低对照(si-NC)组、HG+SNHG14 敲低(si-SNHG14)组、HG+si-SNHG14+miR-101-3p抑制剂(in-miR-101-3p)组、HG+si-SNHG14+SGK1 过表达质粒(pcDNA-SGK1)组.RT-qPCR 检测各组SNHG14、miR-101-3p、SGK1 mRNA表达;CCK-8、克隆形成、免疫荧光以及Western blot检测各组HGMC增殖及纤维化;双荧光素酶和RIP 实验检测miR-101-3p 与SNHG14(或SGK1)关系.结果:DN组患者血清中SNHG14、SGK1 mRNA表达水平较NC组升高,而miR-101-3p表达则降低(P<0.05).与NG组比较,HG组SNHG14 的表达上调,同时细胞增殖活性(OD 值、克隆形成量、Ki67 阳性细胞率)和纤维化标志物(SGK1,α-SMA,FN,Col IV)的表达均显著增强,而miR-101-3p 表达下调(P<0.05).重要的是,与HG+si-NC组相比,敲低SNHG14(HG+si-SNHG14 组)有效逆转了上述HG效应,显著抑制了细胞的增殖和纤维化(P<0.05);而在敲低 SNHG14 的基础上,抑制 miR-101-3p(HG+si-SNHG14+in-miR-101-3p 组)或过表达 SGK1(HG+si-SNHG14+pcDNA-SGK1 组),均能显著逆转因敲低SNHG14 而产生的抗增殖和抗纤维化效应(P<0.05).SNHG14 靶向调控 miR-101-3p/SGK1 轴(P<0.05).结论:lncRNA SNHG14 通过抑制miR-101-3p上调SGK1 表达,促进HG诱导GMC增殖及纤维化进程.
Objective:To explore the effect of LncRNA SNHG14 on high glucose(HG)-induced prolif-eration and fibrosis of glomerular mesangial cells(GMC)by regulating the miR-101-3p/SGK1 axis.Meth-ods:The serum of patients with diabetic nephropathy(DN)(DN group)and healthy people(NC group)were collected.RT-qPCR was used to detect the expression of SNHG14,miR-101-3p,and SGK1 mRNA in serum.Human GMC(HGMC)was cultured and assigned into NG group,HG group,HG+SNHG14 knock-down control(si-NC)group,HG+SNHG14 knockdown(si-SNHG14)group,HG+si-SNHG14+miR-101-3p inhibitor(in-miR-101-3p)group,and HG+si-SNHG14+SGK1 overexpression plasmid(pcDNA-SGK1)group.RT-qPCR was used to detect the expression of SNHG14,miR-101-3p,and SGK1 mRNA in each group.CCK-8,cloning formation,immunofluorescence,and Western blot were used to detect HGMC proliferation and fibrosis in each group.The dual luciferase and RIP assays were used to detect the relation-ship between miR-101-3p and SNHG14(or SGK1).Results:The expression levels of SNHG14 and SGK1 mRNA in serum of patients in DN group were higher than those in the NC group,while the expression of miR-101-3p was lower(P<0.05).Compared with the NG group,SNHG14 expression in the HG group was up-regulated,accompanied by significantly increased cell proliferation activity(OD value,colony formation,Ki67-positive cell rate)and elevated expression of fibrosis markers(SGK1,α-SMA,FN,Col IV),while miR-101-3p expression was downregulated(P<0.05).Importantly,compared with the HG+si-NC group,knockdown of SNHG14(HG+si-SNHG14 group)effectively reversed these HG-induced effects,significantly inhibiting cell proliferation and fibrosis(P<0.05).Furthermore,on the basis of SNHG14 knockdown,either inhibition of miR-101-3p(HG+si-SNHG14+in-miR-101-3p group)or overexpression of SGK1(HG+si-SNHG14+pcDNA-SGK1 group)could significantly reverse the anti-proliferative and anti-fibrotic effects caused by SNHG14 knockdown(P<0.05).SNHG14 targeted and regulated the miR-101-3p/SGK1 axis(P<0.05).Conclusion:LncRNA SNHG14 promotes HG induced proliferation and fibrosis of GMC by inhibiting miR-101-3p and upregulating SGK1.
牛晓静;叶寒露;张利芳;张建
湖北省武汉市中医医院分泌科,湖北 武汉 430014湖北省武汉市中医医院分泌科,湖北 武汉 430014湖北省武汉市中医医院分泌科,湖北 武汉 430014湖北省武汉市中医医院分泌科,湖北 武汉 430014
LncRNA SNHG14miR-101-3p/SGK1轴高糖肾小球系膜细胞纤维化
LncRNA SNHG14miR-101-3p/SGK1 axisHigh glucoseGlomerular mesang-ial cellsFibrosis
《河北医学》 2026 (1)
1-8,8
武汉市中医药科研项目,(编号:WZ24A18)
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