灵杞黄斑颗粒含药血清对兔视网膜色素上皮细胞氧化损伤的保护作用  

Chinese herbal medicine Lingqi Huangban Granule protects retinal pigment epithelial cells against oxidative stress-induced injury in vitro

作　　者：李彩红[1] 邱庆华[2] 吴星伟[2] 宫媛媛[2] 解正高[3] 宋毅[4] 顾青[2] 

机构地区：[1]郑州市中心医院眼科,河南郑州450007 [2]上海交通大学第一人民医院眼科,上海200080 [3]苏北人民医院眼科,江苏扬州225001 [4]上海中医药大学上海市中医医院眼科,上海200072

出　　处：《中西医结合学报》2012年第1期85-90,共6页Journal of Chinese Integrative Medicine

基　　金：上海市科学技术委员会基金资助项目(No.05dz22322)

摘　　要：目的:探讨灵杞黄斑颗粒含药血清对体外培养的兔视网膜色素上皮(retinal pigment epithelium,RPE)细胞氧化损伤的保护作用。方法:运用血清药理学方法制备灵杞黄斑颗粒含药血清,用过氧化氢(500μmol/L)诱导RPE细胞建立体外氧化应激模型。实验分为正常对照组、模型组、空白血清组、低剂量药物血清组和高剂量药物血清组。分别采用原位末端转移酶标记技术(teminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling,TUNEL)和流式细胞仪对各组RPE细胞的凋亡进行检测,并采用蛋白质印迹法检测各组细胞胱天蛋白酶3(caspase-3)及Bcl-XL蛋白表达的变化。结果:流式细胞仪检测结果显示,正常对照组、模型组、空白血清组、低剂量药物血清组、高剂量药物血清组各组凋亡率分别为(4.85±0.26)%、(20.02±1.37)%、(21.84±0.94)%、(13.56±0.55)%、(8.58±0.39)%;含药血清组的凋亡率比模型组明显减少(P<0.05)。TUNEL检测结果显示,凋亡细胞核呈现黄色着染,低剂量药物血清组中散见凋亡细胞,数量明显少于模型组,高剂量药物血清组中凋亡细胞进一步减少。Caspase-3和Bcl-XL表达检测显示,模型组caspase-3蛋白表达上调,高于高剂量药物血清组(P<0.05);而Bcl-XL蛋白表达下调,显著低于低剂量药物血清组和高剂量药物血清组(P<0.01)。结论:灵杞黄斑颗粒含药血清对兔RPE细胞氧化损伤具有一定的保护作用,该作用可能是通过清除自由基,下调caspase-3并上调Bcl-XL的表达实现的。OBJECTIVE： To observe the protective effects of drug-contained serum of Lingqi Huangban Granule （LQHBG）, a compound traditional Chinese herbal medicine, on oxidative stress-induced injury in rabbit retinal pigment epithelial （RPE） cells in vitro. METHODS： The oxidative stress of rabbit RPE cells in vitro was induced with hydrogen peroxide （500 μmol/L） and different concentrations of LQHBG were administered to rats to prepare medicated serum. RPE cells were randomized into normal control group （no hydrogen peroxide）, model group （hydrogen peroxide）, model plus serum group （hydrogen peroxide and 10% control serum）, model plus low-dose LQHBG group （hydrogen peroxide and low-dose LQHBG-medicated serum） and model plus high-dose LQHBG group （hydrogen peroxide and high-dose LQHBG-medicated serum）. Teminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling （TUNEL） assay and flow cytometry （FCM） were used to measure apoptosis of cultured rabbit RPE cells. Protein expressions of caspase-3 and Bcl-XL were observed by Western blot method. RESULTS： FCM results showed that the apoptotic rates of the normal control group, model group, control serum group and serum containing low- and high-dose LQHBG groups were （4.85±0.26） %, （20.02±1.37） %, （21.84±0.94） %, （13.56±0.55） %, and （8.58±0.39） %, respectively; compared with the model group, the apoptotic rates of RPE cells in the low- and high-dose LQHBG groups were obviously reduced in a dose-related manner （P〈0.05）. TUNEL results showed that nuclei of apoptotic cells were stained brown; the number of apoptotic cells in the low- and high-dose LQHBG groups was obviously less than that in the model group. The protein expression of caspase-3 was up-regulated in the model and control serum groups, which was higher than that in the high-dose LQHBG group （P〈0.05）. The protein expression of Bcl-XL was down-regulated in the model and control serum groups, which was lower than that in the lo

关 键 词：中草药 年龄相关性黄斑变性 视网膜色素上皮细胞 氧化应激 血清药理学 凋亡 体外研究 

分 类 号：R285.5[医药卫生—中药学]