1. 附属第一医院内科重症监护室
2. 呼吸病研究所,广东,广州,510080
网络首发:2020-09-11,
纸质出版:2020
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葛珊慧, 张莉珊, 林山, 等. Ghrelin 预处理骨髓间充质干细胞来源上清液对内皮细胞增殖、迁移及凋亡的影响[J]. 中山大学学报(医学科学版), 2020,41(5).
GE Shan⁃hui, ZHANG Li⁃shan, LIN Shan, et al. Effect of Conditioned Medium of Bone Marrow Mesenchymal Stem Cells Pretreated by Ghrelin on Proliferation,Migration and Apoptosis of Endothelial Cells[J]. Journal of Sun Yat-sen University (Medical Sciences), 2020, 41(5).
葛珊慧, 张莉珊, 林山, 等. Ghrelin 预处理骨髓间充质干细胞来源上清液对内皮细胞增殖、迁移及凋亡的影响[J]. 中山大学学报(医学科学版), 2020,41(5). DOI:
GE Shan⁃hui, ZHANG Li⁃shan, LIN Shan, et al. Effect of Conditioned Medium of Bone Marrow Mesenchymal Stem Cells Pretreated by Ghrelin on Proliferation,Migration and Apoptosis of Endothelial Cells[J]. Journal of Sun Yat-sen University (Medical Sciences), 2020, 41(5). DOI:
【目的】研究生长激素释放肽(ghrelin)预处理骨髓间充质干细胞(BMSCs)来源上清液对内皮细胞增殖、迁移及凋亡的影响,并初步探索其作用机制。【方法】全骨髓贴壁分离法提取大鼠BMSCs,利用流式细胞术鉴定 BMSCs 表面分子标记。收集 BMSCs 上清液及高、中、低浓度(100 nmol/L、10 nmol/L、1 nmol/L)ghrelin 预处理BMSCs 24 h 后上清液,以无血清培养基为对照,将上清液分别加入人脐静脉细胞融合细胞(EA.hy926)作用24 h, 利用 CCK8 法检测各组细胞活力,划痕实验评估各组细胞迁移能力。上述各组细胞加入脂多糖处理 24 h 后,用Annexin V 法检测各组细胞凋亡率,Western blot 法检测各组细胞凋亡蛋白bax、caspase3 及β⁃catenin 表达情况。【结果】与未处理 BMSCs 相比,高浓度 ghrelin 预处理 BMSCs 来源上清液能增强内皮细胞活力及迁移能力,差异有统计学意义(P<0.05),流式细胞结果显示,100 nmol/L ghrelin 预处理 BMSCs 组内皮细胞凋亡率(34.51 ± 3.51)%较未处理 BMSCs 组(41.11±1.83)%显著降低,差异有统计学意义(P<0.05)。Western blot 结果显示,100 nmol/L ghrelin 预处理 BMSCs 组较未处理 BMSCs 组凋亡蛋白bax 及 caspase 3 与β⁃catenin 表达下降。【结论】ghrelin 预处理 BMSCs 可以改善内皮细胞功能,其中抗内皮细胞凋亡作用可能与其抑制Wnt/β⁃catenin 通路激活有关。
【Objective】To investigate the impact of conditioned medium derived from bone marrow mesenchymal stem cells(BMSCs)pretreated by ghrelin on proliferation,migration and apoptosis of endothelial cells(EA.hy926)and its underlying mechanism.【Methods】Rat BMSCs were isolated and cultured in vitro by the whole bone marrow adherence method ,and surface markers of BMSCs were identified with flow cytometry. The conditioned medium of BMSCs and BMSCs pretreated by high,moderate and low concentration(100 nmol/L,10 nmol/L and 1 nmol/L)of ghrelin for 24 hours was collected and cocultured with EA.hy926 for 24 hours,and the serumfree medium was considered as the control group. Cell viability was detected by cell counting kit⁃ 8 while migration ability was analyzed by scratch assays. The cell apoptosis rates were measured by Annexin V method after treatment of lipopolysaccharide for 24 hours,and the protein expression of bax,caspase3,and β⁃ catenin were detected by western blot.【Results】Compared with untreated BMSCs group,high concentration of ghrelin pretreated BMSCs group indicated higher cell viability and migration ability (P< 0.05). In light of the result of flow cytometry,endothelial cells apoptosis rate of 100 nmol/L ghrelin pretreated BMSCs group(34.51±3.51%)were remarkably lower than that of untreated BMSCsgroup(P<0.05). As revealed by western blot, expression level of bax,caspase3,and β⁃catenin were reduced in 100 nmol/L ghrelin pretreated BMSCs group compared with untreated BMSCs group(P<0.05).【Conclusion】Endothelial cell function was improved when co⁃cultured in conditioned medium of BMSCs pretreated by ghrelin,and the attenuation of LPS⁃induced apoptosis was related with inhibition of Wnt/β⁃catenin signal path in endothelial cells.
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