银杏叶提取物对瓣膜间质细胞钙化的保护作用
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国家自然科学基金(81900349)


Ginkgo biloba extract EGB761 alleviates aortic valve interstitial cells calcification
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    摘要:

    目的:研究银杏叶提取物(ginkgo biloba extract,EGB761)对华法林诱导的主动脉瓣膜间质细胞(aortic valve interstitial cells,AVIC)钙化及成骨分化的保护作用。方法:分离猪AVIC,将不同浓度的EGB761作用于AVIC,通过茜素红染色、细胞内钙含量以及碱性磷酸酶(alkaline phosphatase,ALP)活性检测观察EGB761对猪AVIC钙化的作用。Real-time PCR和Western blot检测成骨分化相关基因Runt相关转录因子2(runt-related transcription factor 2,Runx2)、Msx2(msh homeobox 2)、骨形态发生蛋白2(bone morphogenetic protein-2,BMP2)和磷酸化Smad1/5的表达。结果:EGB761显著抑制华法林诱导的AVIC钙化,成骨分化相关基因Runx2、Msx2、BMP2的mRNA表达减低,Smad1/5磷酸化及Runx2蛋白表达被抑制。结论:EGB761可能是通过下调Smad1/5磷酸化水平,抑制BMP2/Smad1/5/Runx2信号通路,从而缓解了AVIC钙化和成骨分化。

    Abstract:

    Objective:To investigate the protective effects of ginkgo biloba extract(EGB761) on warfarin-induced calcification and osteogenic differentiation of aortic valve interstitial cell(AVIC). Methods:Porcine AVICs were extracted,and alizarin red staining,intracellular calcium content and alkaline phosphatase activity were detected to analyze the effects of different concentrations of EGB761 on the calcification. The expressions of Runt-related transcription factor 2(Runx2),msh homeobox 2(Msx2),bone morphogenetic protein-2(BMP2)and phosphorylated Smad1/5 were detected by RT-PCR and Western blot. Results:Our results indicated that pAVICs from stimulation with warfarin increased calcium deposition and expression of osteogenic markers(Runx2,Msx2,and BMP2)and promoted phosphorylated-Smad1/5 from the cytoplasm to the nucleus. The addition of EGB761 significantly inhibited phosphorylated-Smad1/5 from the cytoplasm to the nucleus,thus,suppression calcification. Conclusion:EGB761 may reduce the phosphorylation level of Smad1/5 and inhibit the BMP2/Smad1/5/Runx2 signaling pathway,thereby alleviating the calcification and osteogenic differentiation of AVIC.

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刘 静,孔祥清.银杏叶提取物对瓣膜间质细胞钙化的保护作用[J].南京医科大学学报(自然科学版),2021,(10):1440-1446

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  • 收稿日期:2020-11-06
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  • 在线发布日期: 2021-11-05
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