摘要
目的探讨锌指转录因子1(Gli1)对转化生长因子-β1(TGF-β1)诱导的肾小管上皮细胞间质转分化和胶原合成的影响。方法培养肾小管上皮细胞NRK-52E,分为Control、TGF-β1(TGF-β1诱导)、sh-NC+TGF-β1(转染阴性对照shRNA,TGF-β1处理)和sh-Gli1+TGF-β1(转染Gli1 shRNA,TGF-β1处理)组共4组,应用qRT-PCR和Western blot分别检测细胞中Gli1表达水平,ELISA检测细胞培养液上清中纤维连接蛋白(FN)和Ⅰ型胶原酶(ColⅠ)水平,Western blot检测细胞中α-平滑肌肌动蛋白(α-SMA)和Ras相关C3肉毒素底物1(Rac1)蛋白表达水平。结果TGF-β1组肾小管上皮细胞中Gli1 mRNA和蛋白表达量均高于Control组;肾小管上皮细胞中Gli1 mRNA和蛋白表达量,TGF-β1和sh-NC+TGF-β1组均高于sh-Gli1+TGF-β1组;FN和ColⅠ水平,Control组均低于TGF-β1、sh-NC+TGF-β1和sh-Gli1+TGF-β1组,TGF-β1和sh-NC+TGF-β1组均高于sh-Gli1+TGF-β1组;α-SMA和Rac1蛋白表达,Control组均低于TGF-β1、sh-NC+TGF-β1和sh-Gli1+TGF-β1组,TGF-β1和sh-NC+TGF-β1组均高于sh-Gli1+TGF-β1组,差异有统计学意义(P<0.05或P<0.01)。结论下调Gli1减弱TGF-β1条件下肾小管上皮细胞间质转分化和胶原合成水平。
Objective To explore effects of Gli1 on transforming growth factorβ1(TGF-β1)-induced epithelial-mesenchymal transdifferentiation(EMT)and collagen synthesis in renal tubular epithelial cells.Methods Renal tubular epithelial cells NRK-52E were cultured and divided into four groups:control group,TGF-β1 group(TGF-β1 for induction),sh-NC+TGF-β1 group(transfected with negative control shRNA,treated with TGF-β1)and sh-Gli1+TGF-β1 group(transfected with Gli1 shRNA,treated with TGF-β1).Gli1 expression was detected by quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot method respectively.Levels of fibronectin(FN)and type I collagenase(ColⅠ)were detected by enzyme linked immunosorbent assay(ELISA).Western blot method was used to detect protein expressions ofα-smooth muscle actin(α-SMA)and Ras-related C3 botulinum toxin substrate 1(Rac1).Results The mRNA and protein expressions of Gli1 in renal tubular epithelial cells were significantly higher in TGF-β1 group than those in control group,and the expressions in TGF-β1 and sh-NC+TGF-β1 groups were significantly higher than those in sh-Gli1+TGF-β1 group.FN and ColⅠlevels in control group were significantly lower than those in TGF-β1,sh-NC+TGF-β1 and sh-Gli1+TGF-β1 groups,and the levels in TGF-β1 and sh-NC+TGF-β1 groups were significantly higher than those in sh-Gli1+TGF-β1 group.Protein expressions ofα-SMA and Rac1 in control group were significantly lower than those in TGF-β1,sh-NC+TGF-β1 and sh-Gli1+TGF-β1 groups,and the protein expressions in TGF-β1 and sh-NC+TGF-β1 groups were significantly higher than those in sh-Gli1+TGF-β1 group(P<0.05 or P<0.01).Conclusion Down-regulating expression of Gli1 may attenuate TGF-β1-induced EMT and collagen synthesis of renal tubular epithelial cells.
作者
李红微
王志奇
张菊
郭鑫
LI Hong-wei;WANG Zhi-qi;ZHANG Ju;GUO Xin(Department of Nephrology,Liaoyang Central Hospital,Liaoyang,Liaoning 111000,China;Department of Nephrology,People's Hospital of Liaoning Province,Shenyang 111000,China)
出处
《临床误诊误治》
CAS
2021年第7期107-111,共5页
Clinical Misdiagnosis & Mistherapy
基金
辽宁省重点研发计划项目(2019202035)。
关键词
锌指转录因子1
转化生长因子-Β1
肾小管上皮细胞
间质转分化
胶原合成
Glioma associate oncogene homolog 1
Transforming growth factor-β1
Renal tubular epithelial cell
Mesenchymal transdifferentiation
Collagen synthesis
