杞精明目汤含药血清对结膜松弛症MAPK信号通路的影响  

Effect of Qijing Mingmu decoction drug Serum on MAPK Signaling Pathway of Conjunctivochalasis

作　　者：贾元玲 项敏泓[1] 黄丽[1] 李青松[1] 文杭 詹月萍 JIA Yuanling, XIANG Minhong, HUANG Li, et al.(Department of Ophthalmolo-gy, Putuo Hospital affiliated to Shanghai Traditional Chinaese Medicine University, Shanghai200062, China)

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

出　　处：《中国中医眼科杂志》2018年第2期73-80,共8页China Journal of Chinese Ophthalmology

基　　金：上海市医学重点专科-眼科(ZK2015A20);上海中医药大学预算内项目(2015YSN50);上海市普陀区中心医院英才计划(2017202B)

摘　　要：目的探讨杞精明目汤含药血清对结膜松弛症(CCh)成纤维细胞表达丝裂原活化蛋白激酶(MAPK)的影响及其作用机制。方法体外培养结膜松弛症结膜成纤维细胞并鉴定,分为CCh对照组、CCh+20%含药血清组(含药血清组)、CCh+20%无药血清组(无药血清组),予20%的含药或无药血清干预48 h。采用酶联免疫吸附试验(ELISA)检测p38MAPK、c-Jun氨基末端激酶(JNK)、细胞外信号调节激酶(ERK)及其磷酸化水平的光密度值,Western Blot检测p38 MAPK、JNK、ERK蛋白及其磷酸化水平,反转录聚合酶链反应(RT-PCR)检测p38 MAPK、JNK、ERK的m RNA表达,并行统计学分析。结果 ELISA结果显示:含药血清组较CCh对照组p38 MAPK、p-p38 MAPK光密度值明显降低(P<0.05),而无药血清组与CCh对照组差异无统计学意义(P>0.05)。含药血清组较CCh对照组JNK光密度值降低(P<0.05),而无药血清组与CCh对照组差异无统计学意义(P>0.05);含药血清组、无药血清组较CCh对照组p-JNK光密度值下降(P<0.05)。含药血清组较CCh对照组ERK、p-ERK光密度值下降,差异有统计学意义(P<0.05);而无药血清组与CCh对照组差异均无统计学意义(P>0.05)。Western Blot结果显示:含药血清组、无药血清组较CCh对照组p-p38MAPK蛋白表达量明显下降(P<0.05),且20%含药血清的p38 MAPK蛋白磷酸化水平下调更为明显。含药血清组、无药血清组与CCh对照组JNK、p-JNK蛋白表达量差异无统计学意义(P>0.05)。含药血清组、无药血清组与CCh对照组ERK蛋白表达量差异无统计学意义(P>0.05);含药血清组、无药血清组较CCh对照组p-ERK蛋白表达量均明显降低(P<0.05),其中含药血清组的ERK蛋白磷酸化水平下调更为明显。RT-PCR结果显示:无药血清组和含药血清组较CCh对照组p38 MAPK、JNK、ERK的m RNA表达水平均显著下降,差异有统计学意义(P<0.05),且20%含药血清组p38 MAPK、JNK、ERK m RNA水平下调更为明显。结论杞精明目汤含药血清可OBJECTIVE To investigate the effect and mechanism of Qijing Mingmu decoction drug serum on the expression of mitogen activated protein kinase（MAPK） signaling pathway in fibroblasts ofconjunctivochalasis（CCh）. METHODS The conjunctival fibroblasts of CCh were cultured in vitro and identified. The cells were divided into CCh control group, CCh ＋20% drug containing serum group（drug-containing serum group）, CCh＋20% no drug serum group（no-drug serum group）, and20% drug containing or no drug serum were intervened for 48 h. The absorbance values of p38 MAPK, JNK, ERK and their phosphorylation levels were detected by ELISA. The p38 MAPK, JNK and ERK proteins and their phosphorylation levels were detected by Western Blot. The m RNA expressions of p38 MAPK, JNK and ERK were detected by RT-PCR. All the results were statistical analyzed. RESULTS The results of ELISA showed that the drug-containing serum group could significantly reduce the OD value of p38 MAPK and p-p38 MAPK compared with CCh control group（P〈0.05）, but there was no significant difference between no-drug serum group and CCh control group（P〈0.05）. Drug-containing serum group could reduce the OD value of JNK compared with CCh control group（P〈0.05）, but there was no significant difference between no-drug serum group and CCh control group（P〈0.05）. Drug-containing serum group and no-drug serum group could significantly decrease p-JNK OD value（P〈0.05） compared with CCh control group. ERK and p-ERK OD value were decreased in drug-containing serum group compared with CCh control group, and the difference were statistically significant（P〈0.05）, but there was no significant difference between no-drug serum group and CCh control group（P〈0.05）. The results of Western Blot showed that the expression of p-p38 MAPK protein in drug-containing and no-drug serum group were significantly lower than CCh control group（P〈0.05）, and the phosphorylation level of p38 MAPK protein in 20% drug-containing serum group was

关 键 词：结膜松弛症 丝裂原活化蛋白激酶 杞精明目汤 含药血清 成纤维细胞 

分 类 号：R777.3[医药卫生—眼科]