MiR-320c低表达通过抑制TGF-β/Smad通路减轻高糖诱导的肾小管上皮细胞的侵袭及迁移的研究  

Low Expression of MiR-320c Alleviates the Invasion and Migration of High Glucose-induced Renal Tubular Epithelial Cells by Inhibiting TGF-β/Smad Pathway

作　　者：孙衍[1] 渠海[2] 宋祺[1] 申一凡 王俪娟 牛晓红[1] SUN Yan;QU Hai;SONG Qi;SHEN Yi-fan;WANG Li-juan;NIU Xiao-hong(Department of Endocrine,Heji Hospital Affiliated to Changzhi Medical College,Changzhi 046011,Shanxi,China;Department of General Surgery,Heji Hospital Affiliated to Changzhi Medical College,Changzhi 046011,Shanxi,China)

机构地区：[1]长治医学院附属和济医院内分泌科,山西长治046011 [2]长治医学院附属和济医院普通外科,山西长治046011

出　　处：《医学信息》2022年第16期37-42,共6页Journal of Medical Information

基　　金：2019年度山西省高等学校科技创新项目(编号:2019L0700)。

摘　　要：目的通过研究miR-320c对高糖诱导的肾小管上皮细胞侵袭、迁移及TGF-β/Smad通路的影响来探讨miR-320c的作用机制。方法选取人肾小管上皮细胞HK-2进行体外培养,用45 mmol/L浓度的葡萄糖处理24 h,诱导糖尿病肾病肾小管上皮细胞损伤模型设为高糖组(HG),用5.5 mmol/L葡萄糖处理的设为正常对照组(control);inhibitor NC和miR-320c inhibitor分别转染HK-2细胞48 h,并用含有45 mmol/L葡萄糖的培养基培养24 h,分别设为HG+inhibitorNC组和HG+miR-320c inhibitor组;采用实时荧光定量PCR(qRT-PCR)检测miR-320c的表达;光学显微镜观察细胞形态;Transwell侵袭实验检测细胞侵袭;细胞划痕实验检测细胞迁移;蛋白免疫印迹(Western blot)检测TGF-β1及p-Smad2/3蛋白的表达。结果与control组比较,HG组中miR-320c表达显著上调(P<0.001);与inhibitor NC比较,转染miR-320c inhibitor后miR-320c表达显著下调(P<0.01);与control组比较,HG组出现大量的纺锤形细胞,侵袭细胞数及细胞迁移率均显著升高,TGF-β1及p-Smad2/3蛋白表达显著上调(P<0.001);与inhibitor NC组比较,miR-320c inhibitor组纺锤形细胞数量减少,侵袭细胞数及细胞迁移率均显著下调,TGF-β1及p-Smad2/3蛋白表达显著下调(P<0.001)。结论抑制miR-320c可减轻高糖诱导的肾小管上皮细胞的侵袭和迁移,其机制可能与抑制TGF-β/Smad通路的激活有关。Objective To explore the mechanism of miR-320c by studying the effect of miR-320c on the invasion,migration and TGF-β/Smad pathway of renal tubular epithelial cells induced by high glucose.Methods Human renal tubular epithelial cell HK-2 was selected for in vitro culture and treated with 45 mmol/L glucose for 24 h.The injury model of diabetic nephropathy renal tubular epithelial cells was set as high glucose group(HG),and treated with 5.5 mmol/L glucose was set as the normal control group(control).Inhibitor NC and miR-320 c inhibitor were transfected into HK-2 cells for 48 h,respectively,and cultured in medium containing 45 mmol/L glucose for 24 h,respectively,as HG+inhibitor NC group and HG+miR-320 c inhibitor group.The expression of miR-320c was detected by qRT-PCR,and the cell morphology was observed by optical microscope;transwell invasion assay was used to detect cell invasion;cell migration was detected by cell scratch assay;Western blot was used to detect the expression of TGF-β1 and p-Smad2/3.Results Compared with the control group,the expression of miR-320c in the HG group was significantly up-regulated(P<0.001).Compared with inhibitor NC,miR-320c expression was significantly down-regulated after transfection of miR-320c inhibitor(P<0.01).Compared with the control group,a large number of spindle cells appeared in the HG group,the number of invasive cells and cell migration rate were significantly increased,and the expression levels of TGF-β1 and p-Smad2/3 proteins were significantly up-regulated(P<0.001).Compared with inhibitor NC group,the number of spindle cells in miR-320c inhibitor group was decreased,the number of invasive cells and cell migration rate were significantly decreased,and the expression of TGF-β1 and p-Smad2/3 protein was significantly decreased(P<0.001).Conclusion Inhibition of miR-320c can reduce the invasion and migration of renal tubular epithelial cells induced by high glucose,and the mechanism may be related to the inhibition of TGF-β/Smad pathway activation.

关 键 词：MiR-320c 高糖 肾小管上皮细胞 TGF-Β/SMAD 侵袭 迁移 

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