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、1 00、200 μmol/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100 μmol/L建立模型),给予不同浓度(0、10、20、50 μmol/L)BVP处理后(最终采用20 μmol/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.

abstractsObjective:To observe and explore the protective effects ofbreviscapine (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 200 μmogL tBHP (100 μmol/L was chosen to establish models) in the absence or presence of 0,10,20,or 50 μmol/L BVP (20 μmol/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.