筋脉通含药血清对高糖培养背根神经节神经元氧化应激及细胞凋亡的影响  

Effects of Jinmaitong Capsules medicated serum on oxidative stress and apoptosis of rat dorsal root ganglion neurons cultured in high glucose medium

作　　者：孙青[1] 梁晓春[1] 张宏[2] 石玥[1] 张倩[1] 顾蓓[2] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院,北京100730 [2]中国医学科学院基础医学研究所,北京100005

出　　处：《中成药》2013年第7期1390-1395,1448,共7页Chinese Traditional Patent Medicine

基　　金：北京市自然科学基金资助项目(7082077)

摘　　要：目的探讨筋脉通(菟丝子、女贞子、桂枝、延胡索、水蛭、细辛等)含药血清对高糖培养大鼠背根神经节神经元(DRGn)氧化应激及细胞凋亡的影响。方法雄性SD大鼠随机分为3组,分别灌胃筋脉通、维生素C或生理盐水制备含药血清和对照血清。从孕15 d的SD胎鼠背根神经节取材制备细胞悬液。MTT法确定高糖浓度、筋脉通含药血清浓度及指标检测时间点。将体外培养的DRGn分为高糖组、筋脉通组(加入筋脉通含药血清)、维生素C组(加入维生素C含药血清)及正常对照组,培养24 h后,采用荧光探针法检测DRGn中超氧阴离子、线粒体膜电位水平,免疫印迹法+实时荧光定量聚合酶链式反应法检测Bcl-2和Caspase-3的mRNA及其蛋白表达,原位末端标记法检测凋亡细胞。结果与正常对照组相比,高糖组超氧阴离子水平显著升高,Caspase-3 mRNA及其蛋白表达、细胞凋亡率均显著升高,而线粒体膜电位、Bcl-2 mRNA及其蛋白表达显著降低(P<0.01);与高糖组比较,筋脉通组超氧阴离子水平明显降低,Caspase-3 mRNA及其蛋白表达、细胞凋亡率均明显降低,而线粒体膜电位、Bcl-2 mRNA及其蛋白表达显著升高(P<0.01),其作用明显优于维生素C组(P<0.01)。结论筋脉通含药血清可通过抑制高糖所致DRGn氧化应激、减少细胞凋亡起到防治糖尿病周围神经病变的作用。AIM To investigate the effects of Jinmaitong Capsules（Cuscutae Semen,Ligustri lucidi Fructus,Cinnamomi Ramulus,Corydalis Rhizoma,Hirudo,Asari Herba,etc.） medicated serum on oxidative stress and apoptosis of rat dorsal root ganglion neurons（DRGn） cultured in high glucose medium.METHODS Male Sprague-Dawley（SD） rats were divided into normal control group（normal saline）,Jinmaitong Capsules group[Jinmaitong Capsules at dose of 1.31 g/（kg·d）]and vitamin C group[vitamin C at dose of 0.08 g/（kg·d）] to prepare drug medicated serum.Dorsal root ganglia of rat embryo on fifteen days of gestation was dispersed into cell suspension.MTT assay was used to determine the most suitable time point,and the concentrations of glucose and Jinmaitong Capsules medicated serum.DRGn cultured in high glucose medium were divided into high glucose group（cultured with 50 mmol/L glucose medium）,Jinmaitong Capsules group（cultured with Jinmaitong Capsules containing serum） and vitamin C group（cultured with vitamin C containing serum）.After twenty-four hours cultures,the levels of superoxide anion and mitochondrial membrane potential（MMP） were determined by fluorescent probe.The expressions of Bcl-2 and Caspase-3 were detected by Western Blot analysis and qRT-PCR.The apoptotic cells were checked by TUNEL assay.RESULTS Compared with the control group,the levels of superoxide anion and the mRNA and protein levels of Caspase-3 and apoptosis in high glucose group were much higher,but the MMP levels and the mRNA and protein levels of Bcl-2 were much lower（P0.01）.Compared with the high glucose group,the levels of superoxide anion and the mRNA and protein levels of Caspase-3 and apoptosis in Jinmaitong Capsules group were much lower（P0.01）,but the MMP levels and the mRNA and protein levels of Bcl-2 were much higher（P0.01）.Jinmaitong Capsules was superior to vitamin C（P0.01）.CONCLUSION The results indicate that Jinmaitong Capsules medicated serum can antagonize the high glucose-induced oxidative str

关 键 词：筋脉通含药血清 糖尿病周围神经病变 背根神经节神经元 氧化应激 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]