黄连素对脑缺血再灌注小鼠胸腺细胞的保护作用

摘要

Aim:To investigate the protection effects of Bererine (Ber) on the thymocytes of middle cerebral artery occlusion-reperfusion (MCAO/R) injury mouse. Methods: The mouse model of focal cerebral ischemia reperfusion injury was induced by MCAO/R and was given 1.0 h occlusion and 24 h reperfusion. Three groups were randomly divided: Normal group, MCAO/R group which was given the same volume of solvent intraperitoneal injection (I. P. ) , and (MCAO/R + Ber) group, I. P. Ber with a concentration of 5 mg/kg, at 0.5 h ischemia and 12 h reperfusion separatively. After 24 h reperfusion, thymus was removed and thymus index was calculated, then thymocytes were harvested, fluorescence mi-croplate reader and SYTOX Green dying were used to detect the cell activity, used AFM to find the cell morphology changes, and thymocytes early stage of apoptosis was tested by FACS and Calcein/CoCl2 dying or JC-1 staining. Results: MCAO/R led to serious thymus atrophy and a significantly lower thymus index (P＜0. 05). SYTOX Green staining reflected that MCAO/R injury induced higher cell mortality ( P＜0. 05 ). AFM results indicated that notable changes of cell morphology, such as cell height reduced significantly (P＜0. 01) , and rough rate increased remarkably (P＜0. 05 ). Calcein/CoCl2 and JC-1 dying showed that cerebral ischemia/reperfusion resulted in higher apoptosis rate of thymocytes (P＜ 0. 01). On the contrary, Ber improved the thymus atrophy (P＜0. 01) , decreased cell death rate (P＜ 0. 05) , reformed the MCAO/R-induced cell height (P＜0. 01) and average rough (P＜0. 05) , reversed the higher apoptosis rate (P＜0.05). Conclusion: Ber can significantly protect the MCAO/R injury by inhibiting thymocytes apoptosis.%目的:研究黄连素(Ber)对脑缺血再灌注(MCAO/R)小鼠胸腺细胞的保护作用.方法:用线栓法建立小鼠脑缺血再灌注(MCAO/R)模型,缺血1h,再灌注24h,实验动物共分为3组:正常组、手术组和给药组(腹腔注射黄连素质量分数为5 mg/kg),缺血0.5h和再灌注12 h各给药1次.24h后取小鼠胸腺计算其胸腺指数,并取胸腺细胞利用SYTOX Green染色法联合荧光酶标仪检测细胞活性；AFM检测细胞形貌的变化；流式细胞仪结合Calcein/CoCl2或JC-1染色检测早期细胞凋亡率.结果:MCAO/R可引起胸腺明显萎缩；SYTOX Green染色结果显示MCAO/R损伤引起细胞高死亡率(P ＜0.05)；AFM结果表明细胞表面形态发生显著变化(细胞高度差显著减小(P＜0.01)；粗糙度变大(P＜0.05)；Calcein/CoCl2和JC-1结果显示脑缺血再灌注后小鼠胸腺细胞的凋亡率显著增高(P＜0.01),而黄连素能显著改善胸腺的萎缩(P＜0.01),降低细胞的死亡率(P＜0.05),改善MCAO/R引起的细胞高度(P＜0.01)、粗糙度的变化(P＜0.05)和降低细胞的凋亡率(P＜0.05).结论:黄连素可能通过抑制胸腺细胞的凋亡而发挥对脑缺血再灌注的保护作用.