| Lefty-2蛋白对转化生长因子β1诱导的大鼠肾成纤维细胞NRK-49F转分化的逆转作用及其机制 |
| 投稿时间：2015-09-30 修订日期：2015-10-08 点此下载全文 |
| 引用本文：秦聪,张杰,张礼军,吴品.Lefty-2蛋白对转化生长因子β1诱导的大鼠肾成纤维细胞NRK-49F转分化的逆转作用及其机制[J].医学研究杂志,2016,45(4):17-21 |
| DOI：
10.11969/j.issn.1673-548X.2016.04.006 |
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| 基金项目:国家自然科学基金资助项目(81170710) |
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| 中文摘要:目的 观察Lefty-2蛋白对转化生长因子β1(TGF-β1)诱导的肾成纤维细胞(NRK-49F)转分化的逆转作用,并探讨其机制。方法 对照组:NRK-49F不做处理;刺激组:加入TGF-β1(10ng/ml);低剂量治疗组:TGF-β1(10ng/ml)+Lefty-2(10ng/ml);中剂量治疗组:TGF-β1(10ng/ml)+Lefty-2(20ng/ml);高剂量治疗组:TGF-β1(10ng/ml)+Lefty-2(50ng/ml)。通过细胞免疫荧光法观察各组α-平滑肌肌动蛋白(α-SMA)、纤连蛋白(FN)和波形蛋白(Vimentin)的表达水平,通过Western blot法检测各组α-SMA、FN、Vimentin、p-Smad3蛋白、Smad7蛋白的表达水平。结果 细胞免疫荧光结果显示,刺激组α-SMA、FN、Vimentin表达量均显著增加,治疗组较刺激组表达量均明显减少。Western blot法结果显示,治疗组α-SMA、FN、Vimentin的表达量较刺激组均下降(P<0.01),而且呈现剂量效应。刺激组p-Smad3蛋白表达上调,治疗组p-Smad3蛋白的表达量较刺激组明显下调(P<0.01);刺激组Smad7蛋白表达下调,治疗组Smad7蛋白的表达量较刺激组明显上调(P<0.01)。结论 Lefty-2蛋白可逆转TGF-β1诱导的NRK-49F的转分化,可降低α-SMA、FN、Vimentin的表达,可能是通过活化Smad7信号通路,抑制Smad3的磷酸化,从而抑制Smad3信号通路来实现的。 |
| 中文关键词:Lefty-2 TGF-β1 NRK-49F 转分化 |
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| Reversal Effect of Lefty-2 Protein on the Differentiation of Rat Kidney Fibroblasts(NRK-49F) Induced by TGF-β1 and Its Mechanism |
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| Abstract:Objective To observe the reversal effect of Lefty-2 protein on the differentiation of renal fibroblasts (NRK-49F) induced by TGF-β1 and to explore its mechanism. Methods The control group is not treated, and the stimulation group was treated with TGF-β1(10ng/ml), and the low dose treatment group was treated with TGF-β1(10ng/ml)and Lefty-2(10ng/ml), and the middle dose treatment group was treated with TGF-β1(10ng/ml)and Lefty-2(20ng/ml), the high dose treatment group was treated with TGF-β1(10ng/ml)and Lefty-2(50ng/ml). By immunofluorescence, the expression levels of α-SMA, FN, Vimentin were observed and by Western blot, the expression levels of α-SMA, FN, Vimentin, p-smad3 and Smad7 were detected. Results The results showed that the expression of α-SMA, FN and Vimentin in the stimulation group were significantly increased, and the expression of the treatment group was significantly decreased (P<0.01). Western blot results showed that the expression of α-SMA, FN and Vimentin in the treatment group were decreased, and the dose effect relationship was found. The expression of p-Smad3 was up-regulated in the treatment group, the expression of p-Smad3 in the treatment group was significantly lower than that in the stimulation group (P<0.01). The expression of Smad7 in the stimulation group was significantly lower than that in the treatment group (P<0.01). Conclusion Lefty-2 protein can reverse the differentiation of NRK-49F induced by TGF-β1. It can decrease the expression of α-SMA, FN and Vimentin, by activating Smad7 signaling pathway and inhibit the phosphorylation of Smad3 so as to inhibit the Smad3 signaling pathway. |
| keywords:Lefty-2 TGF-β1 NRK-49F Differentiation |
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