灯盏花素抗叔丁基过氧化氢诱导原代大鼠视神经节细胞凋亡  

Protective Effect of Breviscapine on Tert-Butyl Hydroperoxide-induced Primary Retinal Ganglion Cell Apoptosis

作　　者：沈志华 左志琴 彭清华 Zhihua Shen;Zhiqin Zuo;Qinghua Peng(School of Integrated Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;Chinese Medicine School,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学中西医结合学院,长沙410208 [2]湖南中医药大学中医学院,长沙410208

出　　处：《中华眼视光学与视觉科学杂志》2018年第8期454-460,共7页Chinese Journal Of Optometry Ophthalmology And Visual Science

基　　金：湖南省教育厅科学研究项目(17C1221);湖南省研究生创新基金资助项目(CX2016B377、CX2017B434、CX2017B426)。

摘　　要：目的:探讨灯盏花素(BVP)对视神经节细胞的保护作用。方法:实验研究。培养大鼠原代视网膜神经节细胞(PRGCs),采用0、50、100、200μmol/L叔丁基过氧化氢(tBHP)诱导细胞氧化损伤建立细胞模型(最后选择100μmol/L建立模型),给予不同浓度(0、10、20、50μmol/L)BVP处理后(最终采用20μmol/L用于后续研究),检测神经节细胞的改变,免疫印迹法检测细胞凋亡相关因子的改变。采用t检验进行数据处理。结果:与空白组对照相比,经tBHP处理的PRGCs凋亡明显增加,凋亡相关蛋白Bcl-2(P<0.05)和神经标志蛋白Synaptophysin(P<0.01)的表达明显减少。与模型组相比,各治疗组PRGCs成活率明显增加,细胞凋亡减少,Bcl-2和Synaptophysin的表达增加(均P<0.05),而Bcl-2相关X蛋白(Bax)的表达(P>0.05)及CytochromeC的释放(P>0.05)及Caspase-3的剪切则被抑制(P<0.001)。且BVP治疗作用呈剂量依赖性。结论:BVP可以逆转tBHP对视神经节细胞的损伤,减少视神经节细胞的凋亡。Objective:To observe and explore the protective effects of breviscapine(BVP)on tert-butyl hydroperoxide(tBHP)-induced primary retinal ganglion cell(PRGC)apoptosis.Methods:In this experimental study,PRGCs were isolated by immunopanning from retinas of 1-day-old mice and maintained in culture for 3 days.To induce apoptosis,the PRGCs were then exposed to 0,50,100,or 200μmol/L tBHP(100μmol/L was chosen to establish models)in the absence or presence of 0,10,20,or 50μmol/L BVP(20μmol/L was chosen for follow experiments).The PRGCs were identified by immunofluorescence.Expression of Bcl-2,caspase 3,and synaptophysin were assessed by western blotting.Cell apoptosis was detected by the terminal uridine nick-end labeling(TUNEL)assay.Data analysis was performed by t-tests.Results:Compared with the blank control group,the proliferation of PRGCs was significantly inhibited by tBHP(P<0.05).Cell apoptosis increased(P<0.01),and the expressions of Bcl-2 and synaptophysin proteins were significantly down-regulated(P<0.05,P<0.01 respectively).Compared with the tBHP alone,the survival of PRGCs was increased by co-incubation of tBHP with BVP(P<0.01).Cell apoptosis was reduced,along with expressions of Bax(P>0.05)and cleaved-caspase-3(P<0.001).The protein expressions of Bcl-2 and synaptophysin increased(P<0.05).The protective effects of BVP on tBHP-induced PRGC apoptosis were achieved in a concentration dependent manner.Conclusions:BVP can inhibit tBHP-induced PRGC apoptosis.

关 键 词：灯盏花素 氧化应激 视神经节细胞 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]