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侯智为; 李玉莹; 宋海旭; 李佳荫; 邢瑞楠; 刘丹; 刘晶; 闫承慧; 田孝祥
解放军医学杂志, 2019, Volume: 44, Issue: 10Journal Article
R587.1; 目的 探讨糖尿病小鼠(db/db)血清外泌体在体外培养的H9C2心肌细胞损伤中的作用及机制.方法 采用db/db小鼠(n=10)及其对照组小鼠(db/+,n=5)制备糖尿病心肌损伤小鼠模型.提取小鼠血清中的外泌体,定量检测其数量并应用PKH26标记为红色荧光;采用Western blotting检测外泌体相关蛋白及外泌体刺激后H9C2细胞炎性因子的表达情况;采用TUNEL检测细胞凋亡情况;采用Rabla中和抗体进行阻断实验.结果 db/db小鼠血清外泌体数量(30.95×109/ml)明显多于db/+小鼠(10.45×109/ml),差异有统计学意义(P<0.01).体外培养的H9C2细胞能够内吞更多db/db小鼠血清来源的外泌体;应用db/db小鼠血清外泌体干预H9C2细胞可以刺激细胞炎性因子表达明显增加,其中IL-6和IL-1β分别增加6.2倍和2.6倍(P<0.01),且H9C2细胞凋亡明显增多.进一步机制研究发现,Rabla在db/db小鼠血清外泌体中的表达明显增多,应用Rabla中和抗体阻断db/db小鼠外泌体中Rabla表达后可明显抑制H9C2细胞对外泌体的内吞及细胞凋亡.结论 db/db小鼠血清来源的外泌体可诱导体外培养的心肌细胞凋亡及发生炎症反应,可能参与了糖尿病心肌损伤的演进.抑制外泌体分泌或干预其调控分子可能成为糖尿病心肌损伤治疗新的研究靶点.
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