环孢素A滴眼液联合维生素A治疗大鼠干眼症的效果及作用机制

Effect of cyclosporine A eye drops combined with vitamin A on dry eye syndrome in rats and its mechanism

目的观察环孢素A滴眼液联合维生素A治疗大鼠干眼症的效果,并探讨其作用机制。方法将50只SD大鼠随机分为对照组、模型组、环孢素A组、维生素A组、联合用药组,每组10只。除对照组外,其他各组制备干眼症模型,于背部皮下注射氢溴酸东莨菪碱0.6 mg/100μL,4次/d,然后置于通风装置中连续吹风12 h。同时,环孢素A组给予0.05%的环孢素A滴眼,1滴/次,2次/d;维生素A组给予维生素A 200μg/kg灌胃;联合用药组每日同时给予环孢素A和维生素A干预,方法同以上两组。各组均干预10 d,造模期间观察大鼠眼部变化。分别于干预第3、7、10天使用泪液试纸检测泪液分泌量,用荧光素钠试纸检测泪膜破裂时间,用荧光素钠染色法评价角膜损伤程度。实验结束用颈椎脱臼法处死大鼠,取其角膜组织。用HE染色法观察角膜组织病理变化,用Western blotting法检测角膜组织中自杀相关因子(Fas)及其配体(FasL)、B淋巴细胞瘤2(Bcl-2)蛋白表达。结果与模型组比较,环孢素A组、联合用药组泪液分泌量多,泪膜破裂时间长,荧光素钠染色评分高,角膜组织病理损伤轻,Fas、FasL蛋白表达低,Bcl-2蛋白表达高,比较差异均有统计学意义(P均<0.05);且联合用药组以上变化更明显(P均<0.05)。结论环孢素A滴眼液联合维生素A能够显著改善大鼠干眼症症状,其作用机制可能与降低Fas、FasL表达,提高Bcl-2表达有关。

Objective To investigate the effect and mechanism of cyclosporine A eye drops combined with vitamin A on dry eye syndrome in rats.Methods Fifty SD rats were randomly divided into the control group,model group,cyclosporine A group,vitamin A group,and combined drug group,with 10 rats in each group.Except for the control group,rats in the other groups were subcutaneously injected with scopolamine hydrobromide 0.6 mg/100μL,4 times/d,and the rats were placed in a ventilation device for 12 h for continuous blowing.At the same time,the rats in the cyclosporine A group were given 0.05%cyclosporine A eye drops,1 drop/time,twice a day;the rats in the vitamin A group were given vitamin A 200μg/kg by gavage;the rats in the combined drug group were given cyclosporine A and vitamin A for intervention,and the method was the same as the above two groups.Each group was intervened for 10 days,and we observed the changes of the eyes of rats during modeling.On the 3rd,7th,and 10th days of intervention,tear test paper was used to detect tear secretion,tear film rupture time was measured by using sodium fluorescein test paper,and the degree of corneal damage was evaluated by sodium fluorescein staining method.At the end of the experiment,the rats were sacrificed by cervical dislocation,and their corneal tissues were taken.The pathological changes of corneal tissues were observed by HE staining,and the expression levels of suicide-related factor(Fas),suicide-related factor ligand(FasL),and B lymphocyte tumor-2(Bcl-2)protein in the corneal tissues were detected by Western blotting.Results Compared with the model group,the tear secretion was more,the tear film rupture time was longer,and the sodium fluorescein staining scores were higher,the corneal tissue pathological injury was slighter,the protein expression of Fas and FasL proteins was lower,and Bcl-2 protein was higher in the cyclosporine A group and the combined drug group(all P<0.05);and the changes in the combined drug group were more significant(all P<0.05).Conclusions Cyclosporine A eye drops combined with vitamin A can significantly improve the symptoms of dry eye in rats.The mechanism may be related to reducing the expression of Fas and FasL and increasing the expression of Bcl-2.