Alamandine promotes adipogenic differentiation of rat subcutaneous adipose mesenchymal stem cells by acting on MrgD receptors
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摘要:
目的：探讨血管紧张素1-7［angiotensin（1-7），Ang（1-7）］脱羧形成的一种肽Alamandine对大鼠脂肪来源的间充质干细胞（adipose-derived mesenchymal stem cell，ADSC）成脂分化的影响，以及Alamandine调控成脂分化的机制。方法：用不同浓度（0.1、1.0、10.0 μmol/L）的Alamandine处理大鼠ADSC，光镜观察脂滴大小，细胞内甘油三酯、总胆固醇测定，油红O染色法检测成脂分化程度，Western blot检测脂肪分化相关蛋白标志物。确定最适浓度后，检测Alamandine处理大鼠ADSC不同天数的细胞成脂分化程度。血管紧张素Ⅱ（angiotensinII，AngⅡ）和Alamandine共处理大鼠ADSC，相同方法检测成脂分化程度，探讨Alamandine与AngⅡ对ADSC成脂分化的影响。Mas相关G蛋白偶联受体D（mas associated G protein coupled receptor D，MrgD）拮抗剂处理大鼠ADSC，同样的方法检测成脂分化，验证Alamandine促进成脂分化的作用受体。 结果：不同浓度Alamandine处理大鼠ADSC，发现Alamandine促进大鼠ADSC成脂分化，且成明显剂量依赖性。选取10 μmol/L Alamandine处理大鼠ADSC，分别在1、3、6、10 d检测成脂分化指标，随着处理天数增加，成脂分化程度也逐渐增加且在第10天达到顶峰。AngⅡ单处理大鼠ADSC抑制了成脂分化，而且AngⅡ减弱了Alamandine促成脂分化效应。同时MrgD拮抗剂D-pro7抑制了Alamandine对大鼠ADSC的促成脂分化效应。结论：Alamandine通过作用于大鼠ADSC的MrgD起到促进成脂分化的作用。
Abstract:
Objective：This study aims to investigate the effect of Alamandine，a peptide derived from the decarbonization of angiotensin（1-7），on the adipogenic differentiation of adipose-derived mesenchymal stem cell（ADSC）in rats，and the mechanism of Alamandine regulating lipogenic differentiation. Methods：Rat ADSCs were treated with Alamandine at different concentrations（0.1 μmol/L，1.0 μmo/L，10.0 μmol/L），lipid droplets were observed by light microscopy，intracellular triglyceride and total cholesterol were determined，the degree of lipid differentiation was detected by oil red O staining，and the protein associated with adipogenic differentiation was detected by Western blot. After determining the optimal concentration，10 μmol/L Alamandine was used to detect the adipogenic differentiation degree of rat ADSCs in different days. Angiotensin Ⅱ（Ang Ⅱ）and Alamandine dealt with rats ADSCs，same method to detect a concomitant with differentiation，detect the effect of Ang Ⅱ and Alamandine on adipogenic differentiation. Rat ADSCs were treated with Mas associated G protein coupled receptor D（MrgD）antagonist D-pro7，and adipogenic differentiation was detected by the same method，so as to further verify the role of Alamandine in promoting adipogenic differentiation. Results：Alamandine treatment of rat ADSCs with different concentrations showed that Alamandine promoted adipogenic differentiation of rat ADSCs in an obvious dose-dependent manner. ADSCs of rats treated with 10 μmol/L Alamandine were detected at 1，3，6 and 10 days，respectively. With the increase of treatment days，the degree of adipogenic differentiation gradually increased and reached its peak on the 10 th day. Ang Ⅱ single processing rat ADSCs inhibited adipogentic differentiation and the effect of Alamandine contributing to fat differentiation is weakened with Ang Ⅱ. MrgD antagonist D- Pro7 also inhibited the lipid differentiation effect of Alamandine on rat ADSCs. Conclusion：Alamandine promotes lipid differentiation through MrgD on rat ADSCs. This may provide a new way to treat obesity.
