丹七散提取物抗N-甲基-D-天冬氨酸诱导的视神经节细胞凋亡机制  

Protective effects of Danqisan extract on NMDA-injured retinal ganglion cells

作　　者：刘敏[1] 田晗[1] 宁丹[1] 穆云妹 李玉桑[1] 唐和斌[1] 

机构地区：[1]中南民族大学药学院药理学研究室,湖北武汉430074

出　　处：《中国医院药学杂志》2015年第17期1539-1543,共5页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金项目(编号:81202942)

摘　　要：目的:观察丹七散提取物(DQSE)对N-甲基-D-天冬氨酸(NMDA)诱导损伤的大鼠视网膜神经节(RGCL)细胞抗凋亡作用,并探讨其作用机制。方法:Wistar大鼠随机分为对照组、NMDA组、DQSE(3.9,7.8,15.6 g·kg-1)组和阳性对照组(MK801,NMDA抑制剂)。给药7 d后,对照组大鼠玻璃体注射生理盐水,其余组玻璃体注射NMDA制备视网膜损伤模型。给药14 d后,TUNEL法观察RGCL凋亡;免疫组织染色法检测视网膜casepase-3表达分布;Western-blotting法测定视网膜Cleaved casepase-3,-8和-9表达水平。结果:中、高剂量DQSE能显著减少大鼠RGCL的TUNEL阳性表达率(P<0.01,P<0.001)和casepase-3阳性表达率(P<0.01,P<0.001)。DQSE(3.9,7.8,15.6 g·kg-1)剂量依赖性地降低视网膜Cleaved casepase-3,-8,和-9表达水平。结论:DQSE通过下调Cleaved caspase-3蛋白及其上游的Cleaved caspase-8、-9蛋白表达水平产生抗凋亡作用,进而减少RGCL细胞丢失,实现视网膜保护作用。OBJECTIVE To observe therapeutic action of Danqisan extract（DQSE）on retinal ganglion cell layer（RGCL）and investigate underlying mechanism.METHODS Rats were randomly divided into six groups：control group,NMDA group,DQSE groups（3.9,7.8,15.6 g·kg^-1）and positive control groups（MK801,NMDA inhibitors）.After 7 days of medication,rats in control group were treated by intravitreal injection of normal saline,and retinopathy models were established by intravitreal injection of NMDA.After 14 days of medication,apoptosis of RGCL neuron was measured by TUNEL;RGCL caspase-3was tested by immunohistochemistry technique;protein expression levels of Cleaved caspase-3,-8 and-9 were determined by western-blotting assay.RESULTS NMDA-induced Moderate and high dose of DQSE significantly decreased expression levels of TUNEL（P〈0.01,P〈0.001）and casepase-3（P〈0.01,P〈0.001）in RGCL.DQS treatment（3.9,7.8 or 15.6 g·kg^-1）could reverse over expression of Cleaved caspase-3,-8 and-9 in retina of NMDA-treated rats in a dose dependent manner.CONCLUSION DQSE can inhibit NMDA-treated RGCL neuron apoptosis by down regulating levels of caspase-3,-8and-9,thus reduce RGCL loss and protect retina.

关 键 词：丹七散提取物 视网膜神经节细胞 抗凋亡 NMDA Casepase 

分 类 号：R96[医药卫生—药理学]