新型多肽AMPP2在TGF-β1诱导的系膜细胞增殖中的作用及其机制
Effects and mechanism of AMPP2 on mesangial cell proliferation induced by TGF-β1
张琳琳 1赵唐明 1黄婵 1李善文 1甘卫华1
作者信息
- 1. 南京医科大学第二附属医院儿肾科(邮编 210003)
- 折叠
摘要
目的 探究新型多肽AMPP2在转化生长因子β1(TGF-β1)诱导的小鼠系膜细胞增殖中的作用及相关机制.方法 体外用TGF-β1(10µg/L)及AMPP2(10 ng/L)干预系膜细胞,分为Control组、AMPP2组、TGF-β1组及TGF-β1+AMPP2组.CCK-8法检测各组系膜细胞增殖活力;Western blot法检测各组细胞周期蛋白依赖性激酶(CDK)-4、CDK-6、细胞增殖核抗原(PCNA)、α平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原蛋白(COL-Ⅰ)、纤维连接蛋白(FN)及磷酸化的SMAD同源物3/SMAD同源物3(p-SMAD3/SMAD3)的蛋白表达水平;qPCR法检测各组α-SMA、COL-Ⅰ、FN的mRNA表达水平.结果 与Control组相比,TGF-β1组系膜细胞增殖活力增加(P＜0.05),与TGF-β1组相比,TGF-β1+AMPP2组系膜细胞增殖活力下降(P＜0.05);与Control组相比,TGF-β1组CDK-4、CDK-6、PCNA蛋白以及α-SMA、COL-Ⅰ、FN蛋白和mRNA表达水平升高(P＜0.05),与TGF-β1组相比,TGF-β1+AMPP2组上述蛋白及mRNA表达水平均下降(P＜0.05);与Control组相比,TGF-β1组p-SMAD3/SMAD3水平显著升高(P＜0.05),与TGF-β1组相比,TGF-β1+AMPP2组p-SMAD3/SMAD3的蛋白表达量显著降低(P＜0.05).结论 AMPP2可能通过调控TGF-β1/SMAD3通路抑制系膜细胞增殖.
Abstract
Objective To explore the effect and mechanism of anti-mesangial cell-proliferation-peptide 2(AMPP2)on mesangial cell proliferation induced by transforming growth factor β1(TGF-β1).Methods Mesangial cells were cultured in vitro and treated with TGF-β1(10 µg/L)and AMPP2(10 ng/L).According to different intervention factors,mesangial cells were divided into four groups:the control group,the AMPP2 group,the TGF-β1 group and the TGF-β1+AMPP2 group.The proliferation activity of mesangial cells was detected by CCK-8.The relative protein expression of cyclin dependent kinase 4(CDK-4),cyclin dependent kinase 6(CDK-6),proliferating cell nuclear antigen(PCNA),α-smooth muscle actin(α-SMA),collagen-Ⅰ(COL-Ⅰ)and fibronectin(FN)were examined by Western blot assay.The relative mRNA expression of α-SMA,COL-Ⅰ and FN were detected by qPCR.Results Compared with the control group,proliferation activity of mesangial cells was significantly increased in the TGF-β1 group(P＜0.05).The proliferation activity of mesangial cells was markedly decreased in the TGF-β1+AMPP2 group compared with that of the TGF-β1 group(P＜0.05).Compared with the control group,protein levels of CDK-4,CDK-6,PCNA,α-SMA,COL-Ⅰand FN in cells were significantly increased in the TGF-β1 group(P＜0.05),as well as the mRNA levels of α-SMA,COL-Ⅰand FN(P＜0.05).In the TGF-β1+AMPP2 group,the protein and mRNA levels of α-SMA,COL-Ⅰand FN and the protein levels of CDK-4,CDK-6 and PCNA were markedly decreased compared with those of the TGF-β1 group(P＜0.05).Compared with the control group,levels of p-SMAD3/SMAD3 was remarkably upregulated in the TGF-β1 group(P＜0.05),while levels of p-SMAD3/SMAD3 was remarkably downregulated in the TGF-β1+AMPP2 group compared with those of the TGF-β1 group(P＜0.05).Conclusion AMPP2 may inhibit mesangial cell proliferation by regulating TGF-β1/SMAD3 pathway.
关键词
转化生长因子β1/肾小球系膜细胞/细胞增殖/肾炎/紫癜,过敏性/肽类/AMPP2Key words
transforming growth factor beta1/mesangial cells/cell proliferation/nephritis/urpura,Schoenlein-Henoch/peptides/AMPP2引用本文复制引用
基金项目
国家自然科学基金(81970664)
江苏省自然科学基金(BK20191082)
出版年
2024
NETL
NSTL
万方数据