灵杞黄斑颗粒含药血清预处理对人视网膜色素上皮细胞衰老的保护作用

摘要

OBJECTIVE To observe the protective effects of pretreatment with drug-contained serum of "Lingqi Huangban Cranula"(LQHB) on human retinal pigment epithelial (RPE) cell against oxidative stress induced premature senescence in vitro. METHODS Different dosages of drug-contained serum of LQHB were administered. The premature senescence model of cultured human RPE cells was induced with hydrogen peroxide (H2O2) (150μmol/L) and RPE cells were randomized into normal control (NC) group,model (M) group,model + blank serum (MB) group,model + low-dose LQHB (ML) group and model + high-dose LQHB (MH) group. The level of cell activity was measured by the MTS method ,and Flow Cytometer(FCM) method was used to measure the cultured human RPE cellular cycle. Senescence associated beta-Calactose glucoside enzyme Kit is used for the detection of RPE cellular premature senescence (Data were analyzed by statistic software of SAS 9.13). RESULTS Cell activity of the groups which are pretreated with drug-contained serum of LQHB are obviously better than model group, but are lower than normal control group. The results were of significant differences. The cells arrested in G1 phase of groups which are pretreated with drug-contained serum of LQHB are less than the model group .In addition senescence-beta -Galactose glucoside enzyme staining-positive cells of model group is up to 70% ,and normal control group is only 5%. The senescence cell percentages of the groups which are pretreated with drug-contained serum of LQHB are between those of the model group and the normal control group,but no significant statistical differences were found between different dosages. CONCLUSIONS Pretreatment with the drug-contained serum of LQHB has protective effects on human retinal pigment epithelial cellular against the premature senescence induced by oxidative stress injury.%目的 探讨中药灵杞黄斑颗粒(LQHB)含药血清预处理对氧化应激诱导的人视网膜色素上皮(retinal pigment epithelium,RPE)细胞衰老的保护作用.方法 运用血清药理学方法制备不同剂量LQHB含药血清,以过氧化氢(H2O2)(150 μmol/L)诱导建立体外人RPE细胞衰老模型,分为正常对照(NC)组、模型(M)组、模型+空白血清(MB)组、模型+低剂量药物血清(ML)组和模型+高剂量药物血清(MH)组,分别采用MTS法、流式细胞法检测各组RPE细胞的活力和周期,衰老相关-β-半乳糖苷酶试剂盒检测各组RPE细胞的衰老情况.相关统计分析采用SAS 9.13统计学软件.结果 模型组RPE细胞活力较正常对照组明显降低,而LQHB含药血清组细胞活力虽低于正常对照组,但高于模型组.LQHB含药血清组停滞在G1期的RPE细胞较模型组少.模型组衰老相关-β-半乳糖苷酶活性染色阳性的细胞高达70％,而正常对照组仅占5％,LQHB含药血清组衰老细胞百分比介于两者之间,但不同剂量间无统计学意义的差异.结论 LQHB含药血清对氧化应激诱导的人RPE细胞衰老损伤具有一定的保护作用.