淫羊藿总黄酮对干眼症雄兔泪腺中Bax和Bcl-2表达的作用研究

Effect of epimedium total flavonoids on Bax and Bcl-2 expression based on male rabbit of dry eye syndrome

目的观察淫羊藿总黄酮对干眼症雄兔泪腺中Bax和Bcl-2表达的作用。方法随机将80只雄性健康新西兰大耳白兔分为A组(空白组)、B组(手术组)、C组(淫羊藿总黄酮组)、D组(雄激素组),每组20只。A组以生理盐水进行灌胃,B、C、D三组建立干眼症模型后分别以生理盐水灌胃、淫羊藿总黄酮灌胃、丙酸睾酮肌肉注射进行干预。每组根据喂养时间不同又分为A1~D1组(喂养1个月)和A2~D2组(喂养2个月),每组10只。A1~D1组于造模前及造模后2周、4周时,A2~D2组于造模后6周及末次最后一次用药后对雄兔进行泪液分泌实验(SchirmerⅠtest,SIT)和泪膜破裂时间(BUT)检查;A1~D1组和A2~D2组分别在饲养1个月及2个月时处死雄兔,即刻摘取泪腺,应用Western blot检测凋亡相关因子Bax和Bcl-2的蛋白表达。结果SIT、BUT检查结果显示,B、C、D三组雄兔均在造模1个月后形成干眼症。C1组、C2组及D1组、D2组雄兔泪腺组织中的Bax相对含量均较B1组、B2组低,差异均有统计学意义(均为P<0.01);C2组Bax相对含量低于C1组,差异有统计学意义(P<0.05),而C2组与D2组Bax相对含量差异无统计学意义(P>0.05);C1、C2组及D1组、D2组雄兔泪腺组织中Bcl-2相对含量均较B1组、B2组高,差异均有统计学意义(均为P<0.01);C2组Bcl-2表达高于C1组,差异有统计学意义(P<0.05),而C2组与D2组Bcl-2表达差异无统计学意义(P>0.05)。结论淫羊藿总黄酮可提高干眼症雄兔泪腺中Bcl-2的表达,减少Bax的表达,且随用药时间增长其效果逐渐与雄激素相当。淫羊藿总黄酮很可能是通过促进凋亡相关基因Bcl-2、调低Bax的表达达到抑制细胞凋亡的作用。

Objective To observe the effect of epimedium total flavonoids on Bax and Bcl-2 expression based on male rabbit of dry eye syndrome.Methods Eighty male healthy rabbits(2.0-2.5 kg for each rabbit)were divided into blank group(group A),operation group(group B),epimedium total flavonoids group(group C),and androgen group(group D),20 rabbits in each group.Rabbits in the group A were treated with normal saline.After the dry eye models were established in group B,C,and D,group B was treated with normal saline,group C was treated with epimedium total flavonoids,group D was treated with androgen intramuscular injection.Each group was subgrouped into groups A1 to D1(feeding for 1 month)and A2 to D2 group(feeding for 2 months)according to the feeding time,10 animals in each group.In groups A1-D1,the male rabbits were subjected to tear secretion test(Schirmer I test,SIT)and tear film break-up time(BUT)inspections before and at 2 and 4 weeks after the model was created,and in the A2-D2 groups,they were performed at 6 weeks after the model building and after the last medication.Each group of rabbits at the time of the first and second month were sacrificed,with the lacrimal glands taken immediately for extracting the lacrimal gland epithelial cells,and Bax and Bcl-2 protein expression were checked on Western blot.Results SIT and BUT tests showed that dry eye models were established in males of B,C,and D groups after one month.The relative quantity of expression of Bax in lacrimal glands of group C1,C2 and D1,D2 were significantly lower than that of the group B1,B2(all P<0.01).The quantity of expression of Bax in the group C2 was lower than that in the group C1(P<0.05),while the difference was not significant between C1 and D2(P>0.05).The quantity of expression of Bcl-2 in lacrimal glands of group C1,C2 and D1,D2 were significantly higher than that of the group B1,B2(all P<0.01).The quantity of expression of Bax in the group C2 was higher than that in the group C1(P<0.05),while the difference was not significant between C2 and D2(P>0.05).Conclusion Epimedium total flavonoids can promote the expression of Bcl-2 in male rabbit of dry eye syndrome,down-regulate the expression of Bax and the effect gradually became similar to that of androgen.It is possible that the epimedium total flavonoids achieve the inhibition of apoptosis by promoting the expression of Bcl-2 and inhibiting the expression of Bax in lacrimal gland epithelial cells.