滋阴明目丸含药血清对视网膜色素上皮细胞光损伤模型细胞凋亡率及Caspase-1、Caspase-3蛋白表达的影响  

Effects of Serum Containing Ziyin Mingmu Pills(滋阴明目丸) on the Apoptotic Rate and Protein Expression of Caspase-1 and Caspase-3 of Retinal Pigment Epithelial Cells

作　　者：颜家朝[1] 李传课[1] 李波[1] 朱惠安[2] 喻京生[1] 龙辉[2] 陈向东[1] 龙达[1] 吴权龙[1] 易妙 

机构地区：[1]湖南中医药大学第一附属医院,湖南省长沙市韶山中路95号410007 [2]湖南中医药大学

出　　处：《中医杂志》2018年第24期2133-2137,共5页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81303007/H2713);全国名老中医药专家传承工作室[国中医药人教发2013(47)号]

摘　　要：目的探讨滋阴明目丸对视网膜色素上皮(RPE)细胞光损伤后的保护作用及可能机制。方法60只SD大鼠分为滋阴明目丸低、中、高剂量组和空白组,每组15只。滋阴明目丸各剂量组分别予以滋阴明目丸浓度为0. 39、0. 78、1. 56 g/ml的混悬液灌胃,灌胃量均为34 ml/(kg·d),空白组予以生理盐水34 ml/(kg·d)灌胃,各组均给药7天。灌胃结束后制备含药血清,并采用MTT法筛选后续实验的含药血清浓度。实验分为空白组(不加含药血清)、血清组(加含药血清)、模型组(光损伤造模、不加含药血清)及中药组(光损伤造模、加含药血清)。培养24 h后模型组和中药组建立体外RPE细胞光损伤模型。检测各组细胞凋亡率,测定半胱氨酸蛋白酶1 (Caspase-1)、半胱氨酸蛋白酶3 (Caspase-3)蛋白及mRNA表达。结果中剂量血清为最佳给药剂量并用于后续实验。与空白组比较,血清组细胞凋亡率、Caspase-1和Caspase-3蛋白及mRNA表达差异均无统计学意义(P> 0. 05),模型组细胞凋亡率、Caspase-1和Caspase-3蛋白及mRNA表达显著上升(P <0. 05);与模型组比较,中药组细胞凋亡率、Caspase-1和Caspase-3蛋白及mRNA表达显著下降(P <0. 05)。结论滋阴明目丸含药血清能有效抑制RPE细胞光损伤后的细胞凋亡,可能与抑制细胞凋亡因子Caspase-1、Caspase-3蛋白及mRNA表达有关。Objective To observe the effects and possible mechanism of Ziyin Mingmu Pills(滋阴明目丸) on retinal pigment epithelial(RPE) cells after light injury. Methods A total of 60 SD rats were divided into Ziyin Mingmu Pills low,medium and high dose groups and a blank group,with 15 rats in each group. The groups of different Ziyin Mingmu Pills dosages were respectively administered with the suspension of Ziyin Mingmu Pills at a concentration of 0. 39,0. 78,and 1. 56 g/ml. The gavage volume was 34 ml/(kg · d),and the blank group was given34 ml/(kg·d) of normal saline for gavage,each group was administered for 7 days. The drug-containing serum was prepared in the end of the gavage,and the concentration of the drug-containing serum in the subsequent experiment was screened by the MTT method. The selected cells for experiment were divided into a blank group(without drugcontaining serum),serum group(with drug-containing serum),model group(light damage modeling,no drug-containing serum),and Chinese medicine group(light damage modeling with drug-containing serum). After 24 hours of culture,the in vitro RPE cell light damage models of model group and the Chinese medicine group were established.The apoptosis rate of each group was measured,and the expression of caspase-1,caspase-3 and mRNA were measured. Results The medium dose serum was the optimal dose,and it was used in subsequent experiments. Compared with the blank group,the apoptosis rate,expression of Caspase-1 and Caspase-3 protein and mRNA in serum group were not statistically significant(P > 0. 05). The apoptosis rate,Caspase-1 and Caspase-3 protein and mRNA expression in the model group increased significantly(P < 0. 05). Compared with the model group,the apoptosis rate,Caspase-1 and Caspase-3 protein and mRNA expression in the Chinese medicine group were significantly decreased(P < 0. 05). Conclusion Ziyin Mingmu Pills can effectively inhibit the apoptosis of RPE cells after light injury. The mechanism may be related to the inhibition of the expression of Casp

关 键 词：视网膜色素上皮 光损伤 滋阴明目丸 半胱氨酸蛋白酶1 半胱氨酸蛋白酶3 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]