黄芩茎叶黄酮减轻复合Aβ所致大鼠记忆障碍及抑制神经细胞凋亡的调节机制  

Flavonoid from Scutellaria stems and leaves attenuates composited Aβ-induced memory impairment and apoptosis in rats

作　　者：程建军[1] 赵泓翔[1] 郭可[1] 缪红[1] 马帅[1] 高杨[1] 王小青[1] 商亚珍[1] 

机构地区：[1]河北省中医药抗痴呆重点研究室,河北省中药研究与开发重点实验室,承德医学院中药研究所,承德067000

出　　处：《中国新药杂志》2016年第22期2627-2636,共10页Chinese Journal of New Drugs

基　　金：河北省首届百名优秀创新人才支持计划项目(首届);河北省教育厅科学技术重点资助项目(ZD20131022);河北省高等学校重点学科建设项目

摘　　要：目的：探讨黄芩茎叶黄酮（SSF）对β-淀粉样蛋白25-35（Aβ25-35）结合三氯化铝（Al Cl3）及重组人类转化生长因子-β1（RHTGF-β1）（复合Aβ）所致大鼠记忆障碍及抑制神经细胞凋亡的调节机制。方法：SD雄性大鼠手术d 1脑室注射RHTGF-β1,手术d 2开始上午脑室注射Aβ25-35,连续注射14 d,下午脑室注射Al Cl3,连续注射5 d,建立记忆障碍模型,术后d 45用Morris水迷宫进行记忆障碍模型筛选,术后d 49模型成功大鼠随机分为模型组和3个剂量SSF药物组。药物组大鼠分别灌胃35,70和140 mg·kg-1SSF,qd,在给药d 31~37进行Morris水迷宫学习、记忆能力测定。Western blot法测定神经细胞线粒体膜B细胞淋巴瘤/白血病-2（Bcl-2）和Bcl-2相关X蛋白（Bax）及胞液中COX4的蛋白表达;RT-PCR法测定神经细胞胞液中CytC和Caspase-9的mRNA的表达。结果：Morris水迷宫模型筛选结果显示,本实验大鼠模型成功率为98.3%。学习记忆能力测试显示,脑室注射复合Aβ明显引起大鼠记忆获得、记忆保持和记忆再现障碍;使神经细胞线粒体膜上Bcl-2蛋白表达水平明显降低、Bax蛋白表达水平及胞液中Cyto C和Caspase-9 mRNA表达水平都明显增加。而灌胃35,70和140 mg·kg-1SSF不同程度地逆转复合Aβ所致大鼠上述学习记忆障碍及线粒体膜Bcl-2和Bax及胞液中Cyto C和Caspase-9表达的异常改变。线粒体膜和胞液中COX4在各组中变化不明显。结论：SSF能够改善复合Aβ所致大鼠记忆障碍,其作用机制可能是通过抑制神经细胞凋亡和调节线粒体凋亡通路中凋亡因子完成的。Objective： To investigate the effects of flavonoid from Scutellaria stems and leaves （SSF） on memory impairment and regulation mechanism by inhibiting apoptosis in rats induced by amyloid beta protein 25-35 （Aβ25-35） in combination with AlCl3 and RHTGF-131 （composited Aβ）. Methods： The memory impairment model was established by microinjection of RHTGF-β1 into cerebral ventricle of male Sprague Dawley （SD） rats on the first day of operation, accompanied by daily microinjection of Aβ25-35 for 14 d in the morning and AlCl3 for 5 d in the afternoon, respectively from the second day of the operation. The successful model screening was performed for 4 d with Morris water maze since day 45 of operation. On the day 49 of operation, the successful model rats were randomly divided into model group and three SSF-treatment groups. The rats in the drug groups were orally given 35, 70 and 140 mg kg-1 SSF daily. The ability of learning and memory was measured with Morris water maze from day 31 to 37 of drug treatment. Levels of protein expression of Bcl-2 and Bax on mitochondrial membrane and COX4 in cytoplasm were detected with western blotting, and mRNA expressions of Cyt-C and Caspase-9 in cytoplasm were assayed by RT-PCR. Results： The rat model screening by Morris water maze showed a successful model rate of 98. 30% and the ability of learning and memory tests showed that the composited A[3 given by intracerebral ventricle injection obviously caused memory acquirement, memory retention and memory re-extract disorders to rats. The protein expression of Bcl-2 on mitochondrial membrane of neuron dramatically decreased and Bax protein expression and mRNA expression of Cyt-C and Caspase-9 in cytochylema were markedly increased. However, the three doses of SSF 35, 70 and 140 mg kg-1 can differently reversed the above abnormal changes induced by composited Aβ in rats. The changes of COX4 in mitochondrial membrane and cytochylema were not obvious. Conclusion： SSF can ameliorate rats＇ memory impairment

关 键 词：黄芩茎叶黄酮 Β淀粉样蛋白25-35 三氯化铝 人类重组转移因子 记忆 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]