GLP⁃1抑制主动脉瓣膜间质细胞钙化
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国家自然科学基金青年基金(81700340);南京市科技局项目(201715002)


GLP⁃1 inhibits calcification of aortic valve interstitial cells
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    摘要:

    目的:探讨胰高血糖素样肽-1(glucagon-like peptide-1,GLP-1)对主动脉瓣间质细胞(valve stromal cells,VIC)钙化的影响,揭示GLP-1在钙化性主动脉瓣疾病(calcified aortic valve disease,CAVD)发生发展中的保护作用及作用机制。方法:胶原酶消化法分离并培养猪主动脉瓣膜间质细胞,用高钙、高磷法刺激建立主动脉瓣间质细胞钙化模型。采用不同浓度的GLP-1分别干预VICs,通过检测细胞裂解液中钙离子浓度确定最佳浓度。用最佳干预浓度的GLP-1刺激细胞,提取RNA及蛋白,通过RT-PCR以及Western blot分别测定骨桥蛋白(osteopontin,OPN)、Runt相关转录因子2(runt-related transcription factor 2,Runx-2)以及相关信号通路p65蛋白的表达情况。结果:GLP-1作用导致细胞裂解液钙离子浓度降低且最大刺激浓度为200 ng/mL。相比于对照组,GLP-1干预组OPN、Runx-2 mRNA和蛋白表达都降低,p65蛋白表达也降低。结论:GLP-1能够通过抑制p65的表达进而抑制VICs的钙化及其成骨表型分化。

    Abstract:

    Objective:This study aims to investigate the effect of glucagon-like peptide-1(GLP-1)on phenotypic transformation of aortic valve stromal cells(VIC),suggesting the role of GLP-1 in the development of calcified aortic valve disease(CAVD)and the mechanism of action. Methods:The interstitial cells of porcine aortic valve were isolated and cultured by collagenase digestion. The calcification model of aortic valve interstitial cells was established by high calcium and phosphorus stimulation. The calcification model was treated by different concentrations of GLP-1 . The best concentration of GLP-1 was determined by the calcium ion concentration in the cell lysate. The optimal concentration of GLP-1 was used to stimulate the cells,and then RNA and cell proteins were extracted. The expression of osteopontin(OPN),runt-related transcription factor 2(Runx-2)and related signaling pathway p38 were determined by RT-PCR and Western blot. Results:GLP-1 reduced the concentration of calcium ion in cell lysate and the maximum stimulation concentration of GLP-1 was 200 ng/mL. Compared with the control group,mRNAs and proteins of OPN,Runx-2 in GLP-1 intervention group decreased,and the expression of p65 protein also decreased. Conclusion:GLP-1 can inhibit the calcification and osteogenic phenotype differentiation of VICs by inhibiting the expression of p65.

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戴亚伟,周景昕,韩 旭,唐义虎,李明科,吴延虎. GLP⁃1抑制主动脉瓣膜间质细胞钙化[J].南京医科大学学报(自然科学版),2019,(5):664-667

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  • 收稿日期:2018-11-13
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  • 在线发布日期: 2019-05-31
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