杞精明目汤药物血清对结膜松弛症患者球结膜成纤维细胞中基质金属蛋白酶表达的影响  

Effects of qijingmingmu soup on the expression of matrix metalloproteinases in the conjuntival fibroblasts of conjunctivochalasis

作　　者：项敏泓[1] 李轶捷[1] 张兴儒[1] 李青松[1] 韩竹梅[1] 张隆[2] 

机构地区：[1]上海中医药大学附属普陀医院眼科,200062 [2]上海中医药大学实验中心,200062

出　　处：《中华实验眼科杂志》2013年第10期940-943,共4页Chinese Journal Of Experimental Ophthalmology

基　　金：基金项目：上海市高等学校科学技术发展基金项目（09JW41）

摘　　要：背景 结膜松弛症患者球结膜成纤维细胞中基质金属蛋白酶（MMPs）和组织金属蛋白酶抑制剂（TIMPs）的平衡失调,因此寻求一种药物以调节MMPs/TIMPs的平衡对于结膜松弛症的预防和治疗具有重要意义. 目的 研究杞精明目汤药物血清对体外培养的结膜松弛症患者球结膜成纤维细胞中MMPs和TIMPs表达的影响,探讨结膜松弛症的发病机制及杞精明目汤的作用. 方法 24只SD大鼠用杞精明目汤药物灌胃后3d收集腹主动脉血,制备含杞精明目汤药物的血清,以生理盐水灌胃的大鼠血清作为对照.收集接受结膜松弛症切除术患者的球结膜标本,用含质量分数10％胎牛血清的DMEM培养,分别将质量分数20％、15％、10％、5％的药物血清及8 ml/L表皮生长因子（EGF）加入细胞悬液中作用24 h,未加任何药物的结膜成纤维细胞组作为对照组.采用酶联免疫吸附（ELISA）法检测结膜成纤维细胞中MMP1、MMP3、TIMP1和TIMP3的表达（A值）. 结果 培养的细胞呈长梭形,对波形蛋白表达呈阳性.不同质量分数药物血清作用后,人结膜成纤维细胞中MMP1的表达值明显下降,对照组、20％、15％、10％、5％药物血清组及EGF西药组结膜成纤维细胞中MMP1表达值的总体差异有统计学意义（F=466.664,P＜0.05）,其中10％药物血清组MMP1的质量浓度为（9.92±0.14） mg/L,20％药物血清组为（11.87±0.11）mg/L,均明显低于对照组的（16.31±0.10） mg/L,差异均有统计学意义（t=99.974、87.394,P＜0.05）；不同质量浓度药物血清组、EGF西药组和对照组问结膜成纤维细胞中MMP3表达值的总体差异有统计学意义（F=158.168,P＜0.05）,其中20％药物血清组MMP3质量浓度最低,为（3.50±0.03） mg/L,明显低于对照组的（4.44±0.11） mg/L,差异有统计学意义（t=21.991,P＜0.05）.不同质量浓度药物血清组、对照组和EGF西药组间结膜成纤维细胞中TIMP1的表达值差异有统计学意�Background Our previous study determined that expressions of matrix metalloproteinases （MMPs） and tissue matrix metalloproteinase inhibitors （TIMPs）change in the conjuntival fibroblasts of conjunctivochalasis in vitro.To seek a suitable drug is very important in the prevention and treatment of conjunctivochalasis.Objective This study was to explore the effect of qijingmingmu soup on the expressions of MMPs and TIMPs in human conjunctival fibroblasts of conjunctivochalasis.Methods Twenty-four SD rats were randomized into two groups.Qijingmingmu soup was administration gastrically for consecutive 3 days,and normal saline solution was given in the same way in the control group.The blood was collected from aortaventralis and drug serum was prepared.Human conjunctival samples were obtained during the surgery of conjunctivochalasis relaxation and cultured in the DMEM containing 10% fetal bovine serum,20%,15%,10%,5% of drug serum and 8 ml/L epidermal growth factor（EGF） was added into the medium respectively.Enzyme-linked immunosorbent assay（ELISA）was used to detect the expressions of MMP1,MMP3,TIMP1 and TIMP3in conjunctival fibroblasts.Results Cultured cells grew well with the fusiform shape and showed the positive response for vimentin.The expression value of MMP1 （A value）in the cells was declined after administration of qijingmingmu soup.A significant difference was found in the expression of MMP1 among the control group,20%,15%,10%,5% drug serum groups and EGF group（F=466.664,P〈0.05）,and that in the 10% （[9.92±0.14] mg/L） and 20% （[11.87 ±0.11] mg/L） drug serum groups was significantly lowed in comparison with the control group（[16.31±0.10] mg/L）（t=99.974,87.394,P〈0.05）.The expression value of the MMP3in the cells in the various drug serum groups,EGF group and the control group was significantly different（F=158.168,P〈0.05）,with a lower value in the 20% drug serum group compared with the control group （[3.50±0.03] mg/L vs.[4.44 ± 0.11] mg/L） （t =21.991,P �

关 键 词：结膜松弛症 成纤维细胞 中医药 杞精明目汤 基质金属蛋白酶 

分 类 号：R276.7[医药卫生—中医五官科学]