摘要
目的 研究苦参碱对转化生长因子β1(TGF-β1)诱导腹膜间皮细胞上皮间充质转分化(EMT)后转录因子Snail2的影响.方法 采用TGF-β1刺激人腹膜间皮细胞并同时予不同浓度苦参碱干预处理,实验分为空白对照组、TGF-β1(5 ng/ml)诱导组、TGF-β1+0.4 mg/ml苦参碱干预组、TGF-β1+0.6 mg/ml苦参碱干预组、TGF-β1+0.8 mg/ml苦参碱干预组和TGF-β1+1.0 mg/ml苦参碱干预组.实时荧光定量PCR和Western印迹检测Snail2、上皮标志分子E钙黏蛋白(E-cadherin)和间质标志分子α平滑肌肌动蛋白(α-SMA)、纤维连接蛋白(FN)、胶原Ⅲ(ColⅢ)的表达,Western印迹检测Smad2、Smad3和细胞外调节蛋白激酶1/2(ERK1/2)的蛋白磷酸化水平.结果 TGF-β1(5 ng/ml)刺激能上调人腹膜间皮细胞Snail2、α-SMA、FN和ColⅢmRNA和蛋白的表达水平,上调Smad2、Smad3和ERK1/2的蛋白磷酸化水平,下调E-cadherinmRNA和蛋白的表达水平;苦参碱(0.4、0.6、0.8、1.0 mg/ml)干预处理后能下调Snail2和α-SMA、FN和ColⅢmRNA和蛋白的表达水平以及ERK1/2的蛋白磷酸化水平,上调E-cadherin mRNA和蛋白的表达水平.结论 TGF-β1能诱导人腹膜间皮细胞EMT,苦参碱可能通过ERK1/2信号通路下调Snail2的表达水平来抑制TGF-β1诱导的人腹膜间皮细胞EMT.
Objective To study the effect of matrine on the expression of transcription factor Snail2 in peritoneal mesothelial cells epithelial-mesenchymal transition(EMT)induced by transforming growth factor-β1(TGF-β1).Methods Human peritoneal mesothelial cells were stimulated by TGF-β1 and treated with matrine.The experiment was divided into six groups(control group,TGF-β1-induced group(5 ng/ml),TGF-β1+0.4 mg/ml matrine intervention group,TGF-β1+0.6 mg/ml matrine intervention group,TGF-β1+0.8 mg/ml matrine intervention group and TGF-β1+1.0 mg/ml matrine intervention group).The expressions of Snail2,E-cadherin,α-smooth muscle actin(α-SMA),Fibronectin and collagen(Col)Ⅲwere detected by real-time fluorescence quantitative PCR and Western blotting.The protein phosphorylation levels of Smad2,Smad3 and extracellular signal-regulated kinase(ERK)1/2 were detected by Western blotting.Results The mRNA and protein expressions of Snail2,α-SMA,Fibronectin and ColⅢwere up-regulated after being stimulated by TGF-β1(5 ng/ml)in human peritoneal mesothelial cells,while the mRNA and protein expression of E-cadherin was down-regulated.TGF-β1(5 ng/ml)could up-regulate the protein phosphorylation levels of Smad2,Smad3 and ERK1/2.Matrine(0.4,0.6,0.8,1.0 mg/ml)could down-regulate the mRNA and protein expression levels of Snail2,α-SMA,Fibronectin and ColⅢafter being stimulated by TGF-β1 in human peritoneal mesothelial cells.Matrine could down-regulate the protein phosphorylation of ERK1/2,and up-regulate the mRNA and protein expression levels of E-cadherin with treatment of TGF-β1 in human peritoneal mesothelial cells.Conclusions TGF-β1 can induce EMT of human peritoneal mesothelial cells.Matrine may inhibit TGF-β1-induced EMT of human peritoneal mesothelial cells by down-regulating the expression of Snail2 through the ERK1/2 signaling pathway.
作者
李福记
霍冬梅
梁靖梅
段秀萍
廖蕴华
Li Fuji;Huo Dongmei;Liang Jingmei;Duan Xiuping;Liao Yunhua(Department of Nephrology,the First Affiliated Hospital,Guangxi Medical University,Nanning 530021,China)
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2020年第3期221-226,共6页
Chinese Journal of Nephrology
基金
国家自然科学基金(81960149、81660133)。
作者简介
通信作者:廖蕴华,Email:yunhualiao1962@163.com。
