Caspase 3/ROCK1 通路介导高糖诱导的血小板微粒释放。
Caspase 3/ROCK1 pathway mediates high glucose-induced platelet microparticles shedding.
机构信息
Institute of Nephrology, Zhongda Hospital, School of Medicine, Southeast University, Nanjing, 210009, China.
Institute of Nephrology, Zhongda Hospital, School of Medicine, Southeast University, Nanjing, 210009, China.
出版信息
Biochem Biophys Res Commun. 2019 Feb 5;509(2):596-602. doi: 10.1016/j.bbrc.2018.12.166. Epub 2018 Dec 31.
BACKGROUND
Platelet microparticles (PMPs) are closely associated with diabetic macrovascular complications. This study aimed to explore the underlying mechanisms of high glucose-induced PMPs generation.
METHODS
Washed platelets, obtained from the plasma of healthy male Sprague-Dawley rats, were incubated with high glucose. PMPs were isolated using gradient centrifugation and counted by flow cytometry. Expression and activity of ROCK1 and caspase3 were evaluated by real-time PCR, Western blotting, and activity assay kit.
RESULTS
High glucose enhanced PMPs shedding in the presence of collagen. The mRNA and protein levels of ROCK1, but not ROCK2, were increased in platelets incubated with high glucose. Y-27632, an inhibitor of ROCK, blocked the increased PMPs shedding induced by high glucose. Expression and activity of caspase3 were elevated in platelets under the high glucose conditions. Z-DVED-FMK, a caspase3 inhibitor, inhibited ROCK1 activity and decreased the PMPs generation under high glucose.
CONCLUSION
High glucose increased PMPs shedding via caspase3-ROCK1 signal pathway.
背景
血小板微粒（PMPs）与糖尿病大血管并发症密切相关。本研究旨在探讨高糖诱导 PMPs 产生的潜在机制。
方法
从健康雄性 Sprague-Dawley 大鼠血浆中分离出洗涤后的血小板，用高糖孵育。采用梯度离心法分离 PMPs，并用流式细胞术计数。通过实时 PCR、Western blot 和活性测定试剂盒评估 ROCK1 和 caspase3 的表达和活性。
结果
高糖增强了胶原存在下的 PMPs 释放。与高糖孵育的血小板中 ROCK1 的 mRNA 和蛋白水平升高，但 ROCK2 没有升高。ROCK 的抑制剂 Y-27632 阻断了高糖诱导的 PMPs 释放增加。在高糖条件下，血小板中 caspase3 的表达和活性升高。caspase3 抑制剂 Z-DVED-FMK 抑制了 ROCK1 活性并减少了高糖下的 PMPs 生成。
结论
高糖通过 caspase3-ROCK1 信号通路增加 PMPs 的释放。
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