密蒙花滴眼液对去势兔干眼模型泪腺细胞凋亡相关因子Caspase-3、Caspase-8的干预作用

Effect of Buddlejae Officinalis Eyedrops on the Apoptosis-related Factors Caspase-3,Caspase-8 in Lacrimal Glands of Experimental Dry Eye Rabbit Model

目的通过建立去势兔干眼模型,观察密蒙花滴眼液对其泪腺细胞凋亡相关因子Caspase-3、Caspase-8的干预作用。方法将35只新西兰白兔随机分为正常组(A组)、模型组(B组)、0.3、0.5、1.0 mg/mL、1.5、3.0 mg/mL密蒙花滴眼液(C、D、E、F、G组)。A、B组不予处理,C、D、E、F、G组分别予以相应密蒙花滴眼液局部滴眼,3次/日。疗程为4周。造模前、用药前及用药后检测各组基础泪液分泌试验(Schirmer′s test I,SIT)水平,免疫组化检测各组泪腺组织中Caspase-3、Caspase-8的表达情况,Real-time PCR法检测各组泪腺组织中Caspase-3、Caspase-8 mRNA的相对表达情况。结果与A比较,用药前及用药后B组SIT值降低(P<0.01);与B组比较,各治疗组用药后SIT值升高(P<0.01);各治疗组用药后与本组用药前比较,SIT值升高(P<0.01)。与A组比较,B组Caspase-3、Caspase-8表达及mRNA相对表达量升高(P<0.01)。与B组比较,D、F、G组Caspase-3及C、D、E、F、G组Caspase-8表达降低(P<0.01);C、D、E、F、G组Caspase-3及D、E、F、G组Caspase-8 mRNA相对表达量降低(P<0.01)。与C组比较,Caspase-3表达降低(P<0.05);D、E、F、G组Caspase-8 mRNA相对表达量降低(P<0.01)。除D与G组比较外,其他各浓度组间Caspase-3 mRNA相对表达量比较均有统计学意义(P<0.01)。结论密蒙花滴眼液可以通过下调去势兔干眼模型泪腺细胞凋亡因子Caspase-3和Caspase-8的表达,达到维持泪腺基础分泌量和保护泪腺的作用。

Objective To investigate the effects of Buddlejae officinalis eyedrops on the apoptosis-related factors Caspase-3 and Caspase-8 in lacrimal glands of experimental dry eye rabbit model.Methods A total of thirty-five male rabbit were divided into seven study groups,consisting of the control group(group A)and the treatment group(group B,without any treatment),the dry eye rabbit model group treated with different concentration Buddlejae officinalis eyedrops(0.3,0.5,1,1.5 mg/mL and 3 mg/mL)as group C,D,E,F,3 times a day.The course of treatment was 4 weeks.Schirmer’s test I was detected before modeling,before and after treatment.The Caspase-3and Caspase-8 expressions were evaluated by immunohistochemistry and Real-time PCR.Results Compared with group A,the SIT value of the model group was decreased before and after treatment(P<0.01).Compared with group B,the SIT values of each treatment groups were increased after treatment(P<0.01),and the SIT value of each treatment group was higher than that before treatment(P<0.01).Compared with group A,the expression of Caspase-3,Caspase-8 and its mRNA in group B increased(P<0.01).Compared with group B,the expression of Caspase-3 in group D,F,and G decreased(P<0.01),and the expression of Caspase-8 in group C,D,E,F and G was lower(P<0.01).The relative expression of Caspase-3 mRNA in group C,D,E,F,G and the relative expression of Caspase-8 mRNA in group D,E,F,G were decreased(P<0.01).Compared with group C,the expression of Caspase-3 decreased(P<0.05),and the relative expression of Caspase-8 mRNA in group D,E,F and G was lower than that in group C(P<0.01).Except for group D and G,the relative expression of Caspase-3 mRNA in other concentration groups was statistically significant(P<0.01).ConclusionBuddlejae eyedrops can maintain the basic secretion of lacrimal gland and protect lacrimal gland by down-regulating the expression of apoptotic factor Caspase-3 and Caspase-8 in lacrimal gland in castrated rabbit dry eye model,and then,1.5 mg/mL and 3.0 mg/mL Buddlejae eyedrops have better effects.