肉苁蓉毛蕊花糖苷对D-半乳糖诱导PC12神经细胞损伤的保护作用  

Protective Effect of Mullein Glycoside Polysaccharide Extracting from Cistanche on PC12 Nerve-Cell Damage Model Induced by D-galactose

作　　者：苗鑫 张弘 武燕 刘丹丹 张晓菲 李刚 

机构地区：[1]内蒙古医科大学药学院,呼和浩特010110

出　　处：《中国药学杂志》2017年第23期2071-2078,共8页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金资助项目(81260650;81560685);内蒙古科技厅资助项目(KJT14bs805);内蒙古教育厅资助项目(jyt14gnyc03;jyt141133);内蒙古自治区自然科学基金资助项目(kjt14bs805)

摘　　要：目的从调节环腺苷酸/环腺苷酸依赖蛋白激酶/环磷腺苷效应元件结合蛋白(c AMP/PKA/CREB)信号通路水平的角度探讨肉苁蓉毛蕊花糖苷对D-半乳糖诱导PC12神经细胞损伤的保护作用。方法四甲基偶氮唑蓝(MTT)法检测细胞存活率以及提供毛蕊花糖苷剂量选择依据,确定D-半乳糖诱导PC12神经细胞损伤模型的剂量和作用时间,毛蕊花糖苷干预以后,Western Blotting从蛋白水平检测CREB、p-CREB蛋白的表达水平。ELISA法检测c AMP、PKA和脑源性神经营养因子(BDNF)的含量。试剂盒检测超氧化物歧化酶(SOD)、丙二醛(MDA)和乳酸脱氢酶(LDH)的含量。结果 (1)16 g·L^(-1)D-半乳糖暴露PC12细胞40 h后,MTT检测细胞存活率达(46.67±6.59)%,与对照组比较差异具有显著性意义(P<0.05),表明成功建立了细胞衰老模型。(2)与模型组比较,毛蕊花糖苷作用24 h后,p-CREB表达增加(P<0.05),并且有剂量依赖性(r=0.989,P<0.01),PKA、c AMP、BDNF和SOD水平提高(P<0.05),MDA和LDH水平下降(P<0.05)(rMDA=-0.875,P<0.05);(rLDH=-0.834,P<0.05)。阻断剂H-89干预后1 h后,p-CREB表达减少,PKA、c AMP、BDNF和SOD水平显著降低(P<0.05),MDA和LDH水平升高(P<0.05)。结论肉苁蓉中毛蕊花糖苷对D-半乳糖诱导的PC12神经细胞损伤有明显的保护作用,其机制与上调c AMP/PKA/CREB信号通路有关。OBJECTIVE To explore the protective effect of mullein glycoside polysaccharide of Cistanche deserticola Ma on PC12 nerve-cell model induced by D-galactose. METHODS The cell survival rate was determined by MTT assays, which provided the basis for selecting mullein glycoside polysaccharide dose and estimated the dose and action time of D-galactose for inducing PC12 nerve cell damage model. After mullein glycoside polysaccharide incubation of PC12 cells, western blotting was used to detect the levels of CREB and p-CREB protein expression. ELISA Kit was used to detect the levels of cyclic adenosine monophosphate（ cAMP）, cAMP dependent protein kinase （PKA） and brain derived neurotrophic factor（BDNF）. The content of MDA, activities of SOD and LDH were measured by their respective kits. RESULTS （1）After the exposure of the PC12 ceils to 16 g · L- i D-galactose for 40 h, the cell survival rate was （46. 67 ± 6. 59）% , which has a significant difference compared with the control group（ P 〈 0.05）, indicating that successful cell aging model was established. （2）Compared with those in model group, mullein glycoside polysaccharide could significantly increase p-CREB expression in dose-dependent manner（ r = 0. 989, P 〈 0. 01 ）, content of PKA, cAMP, BDNF and SOD and decrease the levels of MDA and LDH （ rMDA = 0. 875, P 〈 0. 05 ） ; （ r LDH = 0, 834, P 〈 0. 05 ）. However, blockers H-89 could significantly decrease p-CREB expression, PKA, cAMP, SOD and BDNF content （P 〈 0. 05） , and increase the levels of MDA and LHD （P 〈 0. 05 ）. CONCLUSION The mullein glycoside polysaccharide of Cistanche deserticola Ma has obvious protective effect on PC12 nerve-cell damage model induced by D-galactose and its mechanism relates to the upregulation of cAMP/PKA/CREB signaling pathways.

关 键 词：肉苁蓉 毛蕊花糖苷 PCI2细胞 环腺苷酸 环腺苷酸依赖蛋白激酶 环磷腺苷效应元件结合蛋白信号通路 

分 类 号：R965[医药卫生—药理学]