β-酪蛋白衍生七肽在高糖水平下可防止NRK-52E细胞发生上皮-间质转化：血管紧张素II-转化生长因子-β1途径的参与
Beta-casomorphin-7 prevents epithelial-mesenchymal transdifferentiation of NRK-52E cells at high glucose level: Involvement of AngII-TGF-β1 pathway.
作者信息
Zhang Wei, Song Shangxin, Liu Fei, Liu Yi, Zhang Yuanshu
机构信息
Key Lab of Animal Physiology and Biochemistry, Ministry of Agriculture, Nanjing Agriculture University, Nanjing, 210095, People's Republic of China; Key Lab of Human Function Genomics Jiangsu Province, Nanjing Medical University, Nanjing, 210029, People's Republic of China.
Key Laboratory of Meat Processing and Quality Control, Ministry of Education, Nanjing Agricultural University, Nanjing 210095, People's Republic of China; Key Laboratory of Agricultural and Animal Products Processing and Quality Control, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, People's Republic of China.
出版信息
Peptides. 2015 Aug;70:37-44. doi: 10.1016/j.peptides.2015.04.002. Epub 2015 Apr 14.
BACKGROUND
Hyperglycemia is the most important risk factor in the progression of renal fibrosis in diabetic kidney. Based on previous studies, β-casomorphin-7 may exert anti-fibrotic activities in diabetic rats. However, the role of β-casomorphin-7 in the pathogenesis of renal tubulointerstitial fibrosis remains unclear. Thus, this study was designed to investigate the protective effect of β-casomorphin-7 on epithelial-mesenchymal transition (EMT) of NRK-52E cells treated under hyperglycemic condition and to explore the possible mechanism.
RESEARCH DESIGN AND METHODS
NRK-52E cells were cultured in high glucose (30 mM) for 3 days. Different concentrations of β-casomorphin-7, naloxone (antagonist of opioid receptor) and losartan (antagonist of angiotensin II type I receptor) were added in the culture. Expression of α-smooth muscle actin (α-SMA), E-cadherin, vimentin and cytokeratin19 mRNA were determined by real-time PCR. Protein levels of E-cadherin and α-SMA were analyzed by Western blotting. The concentrations of angiotensin (Ang) II and transforming growth factor β1 (TGF-β1) in the culture medium were determined.
RESULTS
High glucose-induced up-regulation of vimentin mRNA and α-SMA mRNA and protein were significantly inhibited by β-casomorphin-7. On the contrary, high glucose-induced down-regulation of cytokeratin19 mRNA and E-cad mRNA and protein was significantly reversed by β-casomorphin-7. β-casomorphin-7 significantly alleviate high glucose induced increase of AngII and TGF-β1 in the culture. Moreover, losartan significantly attenuated the expression of TGF-β1 and EMT of NRK-52E cells treated under hyperglycemic condition. But naloxone did not affect the EMT of NRK-52E cells treated by high glucose and β-casomorphin-7.
CONCLUSION
We demonstrate that β-casomorphin-7 has the potential to inhibit high glucose-induced renal proximal tubular EMT partly by modulating AngII-TGF-β1 pathway, but not by opioid receptor.
背景
高血糖是糖尿病肾病肾纤维化进展中最重要的危险因素。基于先前的研究，β-酪蛋白吗啡-7可能在糖尿病大鼠中发挥抗纤维化作用。然而，β-酪蛋白吗啡-7在肾小管间质纤维化发病机制中的作用仍不清楚。因此，本研究旨在探讨β-酪蛋白吗啡-7对高糖条件下处理的NRK-52E细胞上皮-间质转化（EMT）的保护作用，并探索其可能的机制。
研究设计与方法
将NRK-52E细胞在高糖（30 mM）中培养3天。在培养物中加入不同浓度的β-酪蛋白吗啡-7、纳洛酮（阿片受体拮抗剂）和氯沙坦（血管紧张素II 1型受体拮抗剂）。通过实时PCR测定α-平滑肌肌动蛋白（α-SMA）、E-钙黏蛋白、波形蛋白和细胞角蛋白19 mRNA的表达。通过蛋白质印迹分析E-钙黏蛋白和α-SMA的蛋白水平。测定培养基中血管紧张素（Ang）II和转化生长因子β1（TGF-β1）的浓度。
结果
β-酪蛋白吗啡-7显著抑制高糖诱导的波形蛋白mRNA和α-SMA mRNA及蛋白的上调。相反，β-酪蛋白吗啡-7显著逆转高糖诱导的细胞角蛋白19 mRNA和E-钙黏蛋白mRNA及蛋白的下调。β-酪蛋白吗啡-7显著减轻高糖诱导的培养物中AngII和TGF-β1的增加。此外，氯沙坦显著减弱高糖条件下处理的NRK-52E细胞中TGF-β1的表达和EMT。但纳洛酮不影响高糖和β-酪蛋白吗啡-7处理的NRK-52E细胞的EMT。
结论
我们证明，β-酪蛋白吗啡-7有可能部分通过调节AngII-TGF-β1途径而非阿片受体来抑制高糖诱导的肾近端小管EMT。
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