肉苁蓉多糖对D-半乳糖所致急性衰老模型保护作用研究  

In vivo and in vitro studies of protective effect of CDPS on acute aging mouse model induced by D-galactose

作　　者：武燕[1] 张弘[1] 布仁[1] 马慧[1] 苏苗[1] 李刚[1] 

机构地区：[1]内蒙古医科大学药学院,内蒙古呼和浩特010110

出　　处：《中国药理学通报》2017年第7期927-933,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81260650);内蒙古科技厅项目(No KJT14bs805);内蒙古教育厅资助项目(No JYT141133;JYT14qnyco3)

摘　　要：目的通过体内及体外研究肉苁蓉多糖(CDPS)对D-半乳糖所致急性衰老模型的保护作用,并探讨其可能作用机制。方法(1)采用D-半乳糖建立急性衰老小鼠模型,采用Morris水迷宫实验,观察肉苁蓉多糖(25、50、100 mg·kg-1)对衰老所致小鼠学习记忆能力障碍的影响。(2)建立了PC12细胞衰老体外模型,在给予肉苁蓉多糖(150、200 mg·L^(-1))后,采用Western blot检测核蛋白中p-CREB蛋白的表达水平变化。ELISA试剂盒检测环腺苷酸(cA MP)、cA MP依赖蛋白激酶(PKA)和脑源性神经营养因子(BDNF)的含量。此外,在PKA阻断剂H89存在的情况下,检测以上各项指标的变化。(3)建立基于UPLC/Q Exactive质谱检测神经递质谷氨酸、多巴胺和去甲肾上腺素的方法,检测肉苁蓉多糖对于PC12细胞以上神经递质分泌影响。结果(1)体内实验Morris水迷宫实验中,肉苁蓉多糖明显改善D-半乳糖所致衰老模型小鼠学习记忆能力;(2)体外实验中,CDPS给药组作用24h后,可以剂量依赖性地提高细胞核内p-CREB水平(P<0.05),增加PKA及cA MP活性,提高BDNF水平(P<0.05)。阻断剂H-89干预0.5 h后,明显阻断肉苁蓉多糖的这种升高作用。(3)体外实验中,肉苁蓉多糖促进PC12细胞神经递质多巴胺、去甲肾上腺素和谷氨酸分泌。结论肉苁蓉多糖对衰老模型小鼠的学习记忆能力具有明显的改善作用,其机制可能与上调cA MP/PKA/CREB/BDNF信号通路,适度提高兴奋性神经递质有关。Aim To study the protective effect of CDPS on acute aging mouse model induced by D-galactose（D-gal） and its mechanism.Methods（1） The acute aging mouse model was induced by D-gal.After CDPS（25、50、100 mg·kg^-1） treatment,the improving effect on learning and memory in mice was examined in vivo.（2） We also established the aging model on PC12 cells in vitro.After CDPS treatment（150、200 mg· L^-1）,the level of p-CREB in the nucleus was detected by Western blot,and the content of c AMP,PKA and brain derived neurotrophic factor（BDNF） levels were examined by the Elisa kits.Moreover,c AMP,PKA and BDNF were detected in PC12 cells under the condition that H89,the inhibitor of PKA,co-cultured with PC12 cells after CDPS treatment.（3） The UPLC/Q Exactive MS method was developed for determining the concentration of glutamic acid,dopamine and norepinephrine,which secreted in PC12 cells after CDPS treatment.Results（1） In vivo,CDPS significantly improved the memory impairment in aging mice induced by D-gal in the Morris assay.（2） In vitro,CDPS could significantly increase the expression of p-CREB（P〈0.05）,PKA,c AMP and BDNF（P〈0.05）.The H-89 abolished the increase of p-CREB（P〈0.05）,PKA,c AMP and BDNF（P〈0.05） in PC12 aging cells induced by D-gal after CDPS treatment.（3） CDPS increased the release of dopamine,norepinephrine,and glutamate secreted in PC12 cells.Conclusion CDPS could significantly improve the learning and memory ability on aging mouse model in vivo,and reversed the damage in PC12 cells induced by D-gal by activating c AMP/PKA/CREB signal cascade,increase the expression of BDNF,and increasing modestly the release of excitatory neurotransmitter.

关 键 词：肉苁蓉多糖 cAMP/PKA/CREB BDNF 神经递质 抗衰老 D-半乳糖 

分 类 号：R-332[医药卫生] R282.71