Key Laboratory of Basic Pharmacology of Ministry of Education and Guizhou Province, Zunyi Medical University, Zunyi, China.
Joint International Research Laboratory of Ethnomedicine of Ministry of Education, Zunyi Medical University, Zunyi, China.
Phytother Res. 2022 Jul;36(7):2940-2951. doi: 10.1002/ptr.7487. Epub 2022 May 10.
Angiogenesis plays a pivotal role in the recovery of neurological function after ischemia stroke. Herein, we investigated the effect of trilobatin (TLB) on angiogenesis after cerebral ischemia-reperfusion injury (CIRI). The effect of TLB on angiogenesis after CIRI were investigated in mouse brain microvascular endothelium bEnd.3 cells and middle cerebral artery occlusion (MCAO)-induced CIRI rat model. The cell proliferation and angiogenesis were observed using immunofluorescence staining. The cell cycle, expressions of cell cycle-related proteins and SIRT 1-7 were determined by flow cytometry and western blot, respectively. The binding affinity of TLB with SIRT7 was predicted by molecular docking. The results showed that TLB concentration-dependently promoted bEnd.3 cell proportion in the S-phase. TLB significantly increased the protein expressions of SIRT6, SIRT7, and VEGFA, but not affected SIRT1-SIRT5 protein expressions. Moreover, TLB not only dramatically alleviated neurological impairment after CIRI, but also enhanced post-stroke neovascularization and newly formed functional vessels in cerebral ischemic penumbra. Furthermore, TLB up-regulated the protein expressions of CDK4, cyclin D1, VEGFA and its receptor VEGFR-2. Intriguingly, TLB not only directly bound to SIRT7, but also increased SIRT7 expression at day 28. Our findings reveal that TLB promotes cerebral microvascular endothelial cells proliferation, and facilitates angiogenesis after CIRI via mediating SIRT7/VEGFA signaling pathway in rats. Therefore, TLB might be a novel restorative agent to rescue ischemia stroke.
血管生成在缺血性中风后神经功能恢复中起着关键作用。在此，我们研究了三叶海棠素（TLB）对脑缺血再灌注损伤（CIRI）后血管生成的影响。在小鼠脑微血管内皮细胞 bEnd.3 细胞和大脑中动脉闭塞（MCAO）诱导的 CIRI 大鼠模型中研究了 TLB 对 CIRI 后血管生成的影响。通过免疫荧光染色观察细胞增殖和血管生成。通过流式细胞术和 Western blot 分别测定细胞周期、细胞周期相关蛋白的表达和 SIRT1-7。通过分子对接预测 TLB 与 SIRT7 的结合亲和力。结果表明，TLB 浓度依赖性地促进 bEnd.3 细胞进入 S 期。TLB 显著增加了 SIRT6、SIRT7 和 VEGFA 的蛋白表达，但不影响 SIRT1-SIRT5 蛋白表达。此外，TLB 不仅显著减轻 CIRI 后的神经功能损伤，而且增强了脑缺血半影区中风后新生血管和新形成的功能性血管。此外，TLB 上调了 CDK4、cyclin D1、VEGFA 及其受体 VEGFR-2 的蛋白表达。有趣的是，TLB 不仅直接与 SIRT7 结合，而且还在第 28 天增加了 SIRT7 的表达。我们的研究结果表明，TLB 通过调节大鼠 SIRT7/VEGFA 信号通路促进大脑微血管内皮细胞增殖，并促进 CIRI 后血管生成。因此，TLB 可能是一种新的恢复剂，可用于治疗缺血性中风。
