温肾活血方对RCS大鼠视网膜感光细胞凋亡的影响

目的观察温肾活血方对RCS大鼠视网膜感光细胞凋亡的影响。方法纯合子RCS大鼠16只在出生后14 d随机分为2组,即模型组、治疗组,每组8只,另8只SD大鼠作为正常对照组。治疗组予温肾活血方灌胃治疗,而模型组及对照组予蒸馏水灌胃治疗,连续治疗28 d。HE染色观察视网膜各层细胞形态和结构的变化,TUNEL荧光染色观察视网膜外核层细胞凋亡情况。免疫组化荧光染色测定视网膜Caspase-3,Bax,Bcl-2的表达。结果(1)HE染色:治疗后28 d,RCS大鼠各组视网膜外核层较对照组明显变薄,感光细胞数量明显减少,但治疗组较模型组视网膜更厚,感光细胞数更多。(2)TUNEL检测:治疗后28 d,RCS大鼠各组视网膜外核层凋亡细胞计数较对照组明显增多,3组间比较差异有显著统计学意义(F=71.58,P=0.000);与模型组比较,治疗组凋亡细胞计数显著减少(t=4.74,P=0.001)。(3)免疫组化:治疗后28 d,3组间比较,视网膜Caspase-3(F=59.37,P=0.000)、Bcl-2(F=31.35,P=0.000)表达水平差异有统计学意义;与模型组比较,治疗组Caspase-3表达显著减少(t=2.36,P=0.040),Bcl-2表达显著增多(t=3.15,P=0.010),均有统计学意义。结论温肾活血方可以抑制RCS大鼠视网膜感光细胞凋亡,其作用可能与抑制Caspase-3的表达并促进Bcl-2的表达有关。

The protective effect of Wenshen Huoxue recipe on apoptosis of retinal photoreceptor cells in RCS rats

OBJECTIVE To observe the protective effects of Wenshen Huoxue recipe on apoptosis of retinal photoreceptor cells in RCS rats.METHODS On the 14th day after birth,sixteen homozygous RCS rats(half male and half female)were randomly divided into two groups:model control group and treatment group with eight rats in each group,and the other eight SD rats were set as normal control group.The treatment group was given Wenshen Huoxue recipe,and the model group and the normal control group were treated with distilled wate for 28 days.The morphological and structural changes of the retinal cells were observed by HE staining.The apoptosis of cells in the outer retinal nuclear layer was observed by TUNEL fluorescence staining.The expression of Caspase-3,Bax and Bcl-2 in retina was detected by immunohistochemistry.RESULTS(1)HE staining:On the 28th day after treatment,compared with the control group,the outer nuclear layer of RCS rats became thinner and the number of photoreceptor cells in outer nuclear layer decreased significantly.But in the treatment group,the retina was thicker,and the number of photoreceptor cells was more than that in the model group.(2)TUNEL test:On the 28th day after treatment,the apoptotic cells count of the outer nuclear layer of retina in each group of RCS rats was significantly higher than that in the control group and the difference was of statistical significance(F=71.58,P=0.000).But compared with the model group,apoptotic cells count of the outer nuclear layer in the treatment group was decreased significantly(t=4.74,P=0.001).(3)Immunohistochemistry:On the 28th day after treatment,the expression level of Caspase-3(F=59.37,P=0.000),and Bcl-2(F=31.35,P=0.000)in retina of each group were both significantly different.Compared with the model group,the expression of Caspase-3 in the treatment group decreased significantly(t=2.36,P=0.040)while the expression of Bcl-2 increased significantly(t=3.15,P=0.010).CONCLUSIONS Wenshen Huoxue recipe can inhibit the apoptosis of retinal photoreceptor cells in RCS rats,which may be related to inhibiting Caspase-3 expression and promoting Bcl-2 expression.