黄连素对脑缺血再灌注小鼠胸腺细胞的保护作用  

The protection of Bererine on the thymocytes of middle cerebral artery occlusion-reperfusion injury mouse

作　　者：宋兵[1] 王晓东[1] 黄秀艳[1] 曾耀英[1] 

机构地区：[1]暨南大学生命科学技术学院组织移植与免疫实验中心,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2011年第6期584-588,共5页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金："973"国家重大基础研究项目(2006CB504200和2004CB720100);广东省基金项目(2006B36030016);广州市科技局科技攻关重点项目(2006Z-E0091)

摘　　要：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1 h,再灌注24 h,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0.5 h和再灌注12 h各给药1次。24 h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。Aim：To investigate the protection effects of Bererine （Ber） on the thymocytes of middlecerebral artery occlusion-reperfusion （MCAO/R） injury mouse. Methods： The mouse model of focalcerebral ischemia reperfusion injury was induced by MCAO/R and was given 1.0 h occlusion and 24 hreperfusion. Three groups were randomly divided： Normal group, MCAO/R group which was given thesame volume of solvent intraperitoneal injection （ i. p. ）, and （ MCAO/R ＋ Ber） group, i.p. Ber with aconcentration 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 cellmorphology changes, and thymocytes early stage of apoptosis was tested by FACS and Calcein/CoC12 dy-ing or JC-1 staining. Results： MCAO/R led to serious thymus atrophy and a significantly lower thymusindex （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 reducedsignificantly （ P 〈 O. O1 ）, and rough rate increased remarkably （P 〈 O. 05 ）. Calcein/CoC12 and JC-1 dy-ing showed that cerebral ischemia/reperfusion resulted in higher apoptosis rate of thymocytes （P 〈O. O1 ）. On the contrary, Ber improved the thymus atrophy （P 〈0. O1 ）, decreased cell death rate （P 〈0. 05 ）, reformed the MCAO/R-induced cell height （ P 〈 0. 01 ） and average rough （ P 〈 0. 05 ）, reversedthe higher apoptosis rate （P 〈 0.05 ）. Conclusion： Ber can significantly protect the MCAO/R injury byinhibiting thymocytes apoptosis.

关 键 词：黄连素 脑缺血再灌注 胸腺细胞 凋亡 

分 类 号：R285.5[医药卫生—中药学]