转运RNA衍生片段15:31对TGF-β1诱导的小鼠肾小球系膜细胞增殖和炎症反应的影响OACSTPCD
Effects of transfer RNA-derived fragment-15:31 on TGF-β1-induced proliferation and inflammatory response in mouse glomerular mesangial cells
目的 观察转运RNA衍生片段15:31-Arg-CCT-3-1(tRF-15:31)对转化生长因子β1(TGF-β1)诱导的小鼠肾小球系膜细胞增殖和炎症反应的影响.方法 体外培养小鼠肾小球系膜细胞,将细胞分为tRF组、NC组、模型组和对照组.对照组使用正常糖培养基培养细胞.tRF组、NC组、模型组以含10 ng/mL TGF-β1的培养基培养24 h诱导CKD细胞模型;tRF组、NC组分别转染tRF-15:31 mimic和阴性对照.各组干预24 h后,采用CCK-8试剂盒检测细胞增殖能力,采用Western blotting法检测各组细胞中的纤维化指标[(纤维连接蛋白(FN)、Ⅰ型胶原蛋白(Col Ⅰ)、α平滑肌肌动蛋白(α-SMA)]和炎症指标[白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)]蛋白,采用逆转录定量PCR法检测FN、Col Ⅰ、α-SMA、IL-6、TNF-α mRNA,采用ELISA法检测各组细胞培养液上清中的IL-6、TNF-α.结果 模型组增殖活力高于对照组,模型组FN、Col Ⅰ、α-SMA、IL-6、TNF-α蛋白及mRNA表达和培养液上清中IL-6、TNF-α水平高于对照组(P均<0.05);tRF组增殖活力低于NC组,tRF组FN、Col Ⅰ、α-SMA、IL-6、TNF-α蛋白及mRNA表达和培养液上清中IL-6、TNF-α水平低于NC组(P均<0.05).结论 tRF-15:31可抑制TGF-β1诱导的小鼠肾小球系膜细胞增殖,并减少炎症因子分泌.
Objective To investigate the effects of the transfer RNA-derived fragment 15:31-Arg-CCT-3-1(tRF-15:31)on transforming growth factor-β1(TGF-β1)-induced proliferation and inflammatory response of mouse glomerular me-sangial cells.Methods The mouse glomerular mesangial cells were cultured in vitro,and were divided into the follow-ing groups:tRF group,negative control(NC)group,model group and control group.The cells in the control group were cultured using normal glucose medium.Cells in the tRF group,NC group and model group were cultured with medium con-taining 10 ng/mL TGF-β1 for 24 h to induce CKD cell models.The tRF-15:31 mimic and negative control reagent were separately transfected into cells of the tRF group and NC group,respectively.After 24 h of intervention,CCK-8 kit was used to detect cell proliferation.Western blotting was used to detect the fibrosis indexes[fibronectin(FN),collagen type IⅠ(Col Ⅰ),and α-smooth muscle actin(α-SMA)]and inflammation indexes[interleukin-6(IL-6)and tumor necrosis factor α(TNF-α)]in the cells of each group.Quantitative reverse transcription PCR was used to detect FN,Col Ⅰ,α-SMA,IL-6 and TNF-α mRNA,and ELISA was used to detect IL-6 and TNF-α in cell culture medium supernatant of each group.Results Compared with the control group,cell proliferation viability was significantly enhanced and the rel-ative mRNA and protein expression levels of FN,Col Ⅰ,α-SMA,IL-6,and TNF-α were higher in the model group(all P<0.05).The proliferative activity was lower,and the protein and mRNA expression levels of FN,Col Ⅰ,α-SMA,IL-6,TNF-α,and the levels of IL-6 and TNF-α in culture medium supernatant were lower in the tRF group than in the NC group(all P<0.05).Conclusion The tRF-15:31 can inhibit the proliferation of mouse mesangial cells induced by TGF-β1 and reduce the secretion of inflammatory factors.
夏梓桓;单智杰;乔云阳;张爱青
南京医科大学儿科学院南京医科大学第四附属医院儿科
临床医学
慢性肾脏病肾小球系膜细胞转运RNA衍生片段细胞增殖炎症反应
chronic kidney diseasemesangial cellstransfer RNA-derived fragmentscell proliferationinflam-mation
《山东医药》 2024 (020)
30-34 / 5
江苏省高等学校大学生创新创业训练计划项目创新训练重点项目(202210312038Z);江苏省南京市卫生科技发展专项资金项目(YKK23209).
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