[1]孙 衍,渠 海,宋 祺,等.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研究[J].医学信息,2022,35(16):37-42.[doi:10.3969/j.issn.1006-1959.2022.16.008]
　SUN Yan,QU Hai,SONG Qi,et al.Low Expression of MiR-320c Alleviates the Invasion and Migration of High Glucose-induced Renal Tubular Epithelial Cells by Inhibiting TGF-β/Smad Pathway[J].Journal of Medical Information,2022,35(16):37-42.[doi:10.3969/j.issn.1006-1959.2022.16.008]
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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研究()
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医学信息[ISSN:1006-1959/CN:61-1278/R]

卷:
35卷
期数:
2022年16期
页码:
37-42
栏目:
论著
出版日期:
2022-08-15

文章信息/Info

Title:
Low Expression of MiR-320c Alleviates the Invasion and Migration of High Glucose-induced Renal Tubular Epithelial Cells by Inhibiting TGF-β/Smad Pathway
文章编号:
1006-1959（2022）16-0037-06
作者:
孙 衍; 渠 海; 宋 祺; 等
（长治医学院附属和济医院内分泌科1，普通外科2，山西 长治 046011）
Author(s):
SUN Yan; QU Hai; SONG Qi; et al.
(Department of Endocrine1,Department of General Surgery2,Heji Hospital Affiliated to Changzhi Medical College,Changzhi 046011,Shanxi,China)
关键词:
MiR-320c; 高糖; 肾小管上皮细胞; TGF-β/Smad; 侵袭; 迁移
Keywords:
MiR-320c; High glucose; Renal tubular epithelial cells; TGF-β/Smad; Invasion; Migration
分类号:
R587.2；R692.9
DOI:
10.3969/j.issn.1006-1959.2022.16.008
文献标志码:
A
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。
Abstract:
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.

参考文献/References:

[1]Selby NM,Taal MW.An updated overview of diabetic nephropathy:Diagnosis, prognosis, treatment goals and latest guidelines[J].Diabetes,Obesity & Metabolism,2020,22 Suppl 1:3-15.[2]Kramer A,Pippias M,Noordzij M,et al.The European Renal Association-European Dialysis and Transplant Association （ERA-EDTA） Registry Annual Report 2016:a summary[J].Clinical Kidney Journal,2019,12(5):702-720.[3]Stanton RC.Diabetic Kidney Disease and Hypertension[J].Experimental and Clinical Endocrinology & Diabetes,2016,124(2):93-98.[4]Maqbool M,Cooper ME,Jandeleit-Dahm KAM.Cardiovascular Disease and Diabetic Kidney Disease[J].Seminars in Nephrology,2018,38(3):217-232.[5]Lu TX,Rothenberg ME.MicroRNA[J].The Journal of Allergy and Clinical Immunology,2018,141(4):1202-1207.[6]Dai Y,Guo M,Jiang L,et al.Network pharmacology-based identification of miRNA expression of Astragalus membranaceus in the treatment of diabetic nephropathy[J].Medicine,2022,101(5):e28747.[7]Yarahmadi A,Shahrokhi SZ,Mostafavi-Pour Z,et al.MicroRNAs in diabetic nephropathy: From molecular mechanisms to new therapeutic targets of treatment[J].Biochemical Pharmacology,2021,189:114301.[8]Li Y,Huang D,Zheng L,et al.Effect of microRNA-141 on the development of diabetic nephropathy through regulating AKT/AMPK signaling pathway by targeting insulin receptor substrate 2[J].Journal of Cellular Biochemistry,2018,120(5):8008-8015.[9]Hong Y,Wang J,Zhang L,et al.Plasma miR-193a-3p can be a potential biomarker for the diagnosis of diabetic nephropathy[J].Annals of Clinical Biochemistry,2021,58(2):141-148.[10]郭红,赵芳芳.miR-320c调控KIF14对高糖诱导的足细胞损伤及nephrin、podocin表达的影响[J].中国中西医结合肾病杂志,2020,21(9):805-809.[11]Li R,Guo Y,Zhang Y,et al.Salidroside Ameliorates Renal Interstitial Fibrosis by Inhibiting the TLR4/NF-κB and MAPK Signaling Pathways[J].International Journal of Molecular Sciences,2019,20(5):1103.[12]Nieto MA,Huang RY,Jackson RA,et al.EMT: 2016[J].Cell,2016,166(1):21-45.[13]Liu L,Wang Y,Yan R,et al.BMP-7 inhibits renal fibrosis in diabetic nephropathy via miR-21 downregulation[J].Life Sciences,2019,238:116957.[14]Guo J.Effect of miR-21 on Renal Fibrosis Induced by Nano-SiO2 in Diabetic Nephropathy Rats via PTEN/AKT Pathway[J].Journal of Nanoscience and Nanotechnology,2021,21(2):1079-1084.[15]Xue M,Li Y,Hu F,et al.High glucose up-regulates microRNA-34a-5p to aggravate fibrosis by targetingSIRT1 in HK-2 cells[J].Biochemical and Biophysical Research Communications,2018,498(1):38-44.[16]Wang J,Wang G,Liang Y,et al.Expression Profiling and Clinical Significance of Plasma MicroRNAs in Diabetic Nephropathy[J].Journal of Diabetes Research,2019,2019:5204394.[17]Gholaminejad A,Abdul Tehrani H,Gholami Fesharaki M.Identification of candidate microRNA biomarkers in diabetic nephropathy:a meta-analysis of profiling studies[J].Journal of Nephrology,2018,31(6):813-831.[18]Cai S,Liu J,Ma Q,et al.Coptis inhibited epithelial-mesenchymal transition and fibrogenesis of diabetic nephropathy through lncRNA CLYBL-AS2-miR-204-5p-SNAI1 axis[J].Journal of Drug Targeting,2020,28(9):939-948.[19]Hu L,Ding M,He W.Emerging Therapeutic Strategies for Attenuating Tubular EMT and Kidney Fibrosis by Targeting Wnt/β-Catenin Signaling[J].Frontiers in Pharmacology,2021,12:830340.[20]Liu D,Liu F,Li Z,et al.HNRNPA1-mediated exosomal sorting of miR-483-5p out of renal tubular epithelial cells promotes the progression of diabetic nephropathy-induced renal interstitial fibrosis[J].Cell Death & Disease,2021,12(3):255.[21]Zang L,Gao F,Huang A,et al.Icariin inhibits epithelial mesenchymal transition of renal tubular epithelial cells via regulating the miR-122-5p/FOXP2 axis in diabetic nephropathy rats[J].Journal of Pharmacological Sciences,2022,148(2):204-213.[22]Yang Y,Zhou J,Li WH,et al.LncRNA NEAT1 regulated diabetic retinal epithelial-mesenchymal transition through regulating miR-204/SOX4 axis[J].PeerJ,2021,9:e11817.[23]Zhao X,Dai L,Yue Q,et al.MiR-195 inhibits migration, invasion and epithelial-mesenchymal transition (EMT) of endometrial carcinoma cells by targeting SOX4[J].Journal of Biosciences,2019,44(6):146.[24]Wang X,Wei P,Yang L,et al.MicroRNA-20a-5p regulates the epithelial-mesenchymal transition of human hepatocellular carcinoma by targeting RUNX3[J].Chin Med J (Engl),2022 Feb 8,Epub ahead of print.[25]Quan KY,Yap CG,Jahan NK,et al.Review of early circulating biomolecules associated with diabetes nephropathy - Ideal candidates for early biomarker array test for DN[J].Diabetes Research and Clinical Practice,2021,182:109122.[26]Lin HC,Paul CR,Kuo CH,et al.Glycyrrhiza uralensis root extract ameliorates high glucose-induced renal proximal tubular fibrosis by attenuating tubular epithelial-myofibroblast transdifferentiation by targeting TGF-β1/Smad/Stat3 pathway[J].Journal of Food Biochemistry,2022:e14041.[27]Dong L,Yu L,Zhong J.Histone lysine-specific demethylase 1 induced renal fibrosis via decreasing sirtuin 3 expression and activating TGF-β1/Smad3 pathway in diabetic nephropathy[J].Diabetology & Metabolic Syndrome,2022,14(1):2.[28]Wu W,Wang Y,Li H,et al.Buyang Huanwu Decoction protects against STZ-induced diabetic nephropathy by inhibiting TGF-β/Smad3 signaling-mediated renal fibrosis and inflammation[J].Chinese Medicine,2021,16(1):118.[29]Liang L,Li S,Liu H,et al.Blood glucose control contributes to protein stability of Ski-related novel protein N in a rat model of diabetes[J].Experimental and Therapeutic Medicine,2021,22(5):1341.[30]Liu Q,Lv S,Liu J,et al.Mesenchymal stem cells modified with angiotensin-converting enzyme 2 are superior for amelioration of glomerular fibrosis in diabetic nephropathy[J].Diabetes Research and Clinical Practice,2020,162:108093.[31]Zhang J,Zhang L,Zha D,et al.Inhibition of miRNA-135a-5p ameliorates TGF-β1-induced human renal fibrosis by targeting SIRT1 in diabetic nephropathy[J].International Journal of Molecular Medicine,2020,46(3):1063-1073.[32]张永,史秀岩,罗昌霞,等.miR-346在糖尿病肾病TGF-β介导的Smad信号通路中的作用[J].湖北医药学院学报2016,35(3):240-245,250.[33]何其英.基于TGF-β1-Smad2/3探讨芪地糖肾颗粒防治糖尿病肾病肾脏纤维化的研究[D]北京:北京中医药大学,2018.

更新日期/Last Update: 1900-01-01