淫羊藿苷和齐墩果酸协同地塞米松对糖皮质激素敏感型/抵抗型细胞内信号通路蛋白的影响  

Regulatory effect of icariin and oleanolic acid combined with dexamethasone on JAKs/STATs signaling pathway in hormone-resistant CEM-C1 cells

作　　者：唐秀凤[1] 高莹莹 李晓曦[1] 陈宇恒 于萍[1] 许利平[1] 刘仁慧[1] TANG Xiu-feng;GAO Ying-ying;LI Xiao-xi;CHEN Yu-heng;YU Ping;XU Li-ping;LIU Ren-hui(Beijing Key Lab of TCM Collateral Disease Theory Research,School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学中医药学院及中医络病研究北京市重点实验室,北京医学博士研究生100060

出　　处：《医学研究生学报》2018年第9期910-915,共6页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81673993;81373814);北京中医药薪火传承"3+3"工程二室一站建设项目(2012-SZC-42);北京市属高等学校高层次人才引进与培养计划项目(CIT&TCD201504097)

摘　　要：目的淫羊藿、女贞子协同糖皮质激素(GC)治疗哮喘的作用机制与调节哮喘大鼠肺组织中与Th1/Th2平衡密切相关的Janus激酶/信号转导及转录因子通路(JAKs/STATs)通路密切相关。文章研究淫羊藿苷、齐墩果酸协同地塞米松对GC敏感型CEM-C7细胞与GC抵抗型CEM-C1细胞内JAKs/STATs蛋白表达的影响。方法 CEM-C7和CEM-C1细胞株,分别分为6组,包括对照组(完全培养基培养)、地塞米松组(地塞米松10-6mol/L)、淫羊藿苷组(淫羊藿苷100 mg/m L)、齐墩果酸组(齐墩果酸100 mg/m L)、淫羊藿苷+齐墩果酸合用组(淫羊藿苷50 mg/m L、齐墩果酸50 mg/m L)、联合组(地塞米松10-6mol/L+淫羊藿苷50 mg/m L+齐墩果酸50 mg/m L)。给予相应的药物作用24 h后,Western blot法检测细胞内JAKs(JAK1、JAK2)及STATs(STAT1、STAT3、STAT5、STAT6)蛋白表达。结果对CEM-C1细胞的影响:对照组、地塞米松组、淫羊藿苷组、齐墩果酸组、淫羊藿苷+齐墩果酸组、联合组细胞中JAK1表达量分别为0.22±0.01、0.24±0.01、0.23±0.01、0.24±0.01、0.22±0.01、0.18±0.01,JAK2表达量分别为0.23±0.01、0.24±0.01、0.22±0.01、0.23±0.01、0.21±0.01、0.19±0.01;淫羊藿苷+齐墩果酸组、联合组JAK1、JAK2蛋白表达均较地塞米松组显著下调(P<0.01);淫羊藿苷组、齐墩果酸组、联合组较淫羊藿苷+齐墩果酸组降低(均P<0.01)。对照组、地塞米松组、淫羊藿苷组、齐墩果酸组、淫羊藿苷+齐墩果酸组、联合组STAT1的表达量分别为0.23±0.01、0.23±0.01、0.23±0.01、0.23±0.01、0.18±0.01、0.17±0.01,STAT3的表达量分别为0.23±0.01、0.22±0.01、0.22±0.01、0.23±0.01、0.20±0.02、0.16±0.01;淫羊藿苷+齐墩果酸组、联合组STAT1和STAT3蛋白表达均较地塞米松组显著下调(P<0.01);与淫羊藿苷+齐墩果酸组比较,联合组STAT3蛋白表达进一步下调(P<0.01)。对照组、地塞米松组、淫羊藿苷组、齐墩果酸组、淫羊藿苷+齐墩果酸组、联合组STAT5的表Objective The mechanisms of epimedium and Ligustrum Lucidum with glucocorticoid(GC)acting on asthma are closely related to the regulation of the JAKs/STATs pathway associated with the Th1/Th2 balance in the lung tissue of the asthmatic rats.This study aimed to investigate the synergistic effect of icariin and oleanolic acid with dexamethasone on the protein expressions of JAKs/STATs in GC-sensitive CEM-C7 and GC-resistant CEM-C1 cells.Methods We divided CEM-C7 and CEM-C1 cells into groups A(complete culture medium control),B(dexamethasone at 10-6 mol/L),C(icarrin at 100 mg/mL),D(oleanolic acid at 100 mg/mL),E(icarrin+oleanolic acid both at 50 mg/mL),and F(icariin+oleanolic acid+dexamethasone at 50 mg/mL,50 mg/mL and 10-6 mol/L,respectively),and treated them with corresponding agents for 24 hours.Then,we determined the protein expressions of JAKs(JAK1 and JAK2)and STATs(STAT1,STAT3,STAT5,and STAT6)in the CEM-C7 and CEM-C1 cells of different groups by Western blot.Results The protein expressions of JAK1 and JAK2 in the CEM-C1 cells were 0.22±0.01 and 0.23±0.01 in group A,0.24±0.01 and 0.24±0.01 in group B,0.23±0.01 and 0.22±0.01 in group C,0.24±0.01 and 0.23±0.01 in group D,0.22±0.01 and 0.21±0.01 in group E,and 0.18±0.01 and 0.19±0.01 in group F,both significantly lower in groups E and F than in B(P<0.01),and in groups C,D and F than in E(P<0.01).The expressions of STAT1 and STAT3 proteins were 0.23±0.01 and 0.23±0.01 in group A,0.23±0.01 and 0.22±0.01 in group B,0.23±0.01 and 0.22±0.01 in group C,0.23±0.01 and 0.23±0.01 in group D,0.18±0.01 and 0.20±0.02 in group E,and 0.17±0.01 and 0.16±0.01 in group F,both remarkably lower in groups E and F than in B(P<0.01),and that of STAT3 even lower in F than in E(P<0.01).The expressions of STAT5 and STAT6 were 0.24±0.01 and 0.24±0.01 in group A,0.23±0.01 and 0.23±0.02 in group B,0.23±0.01 and 0.24±0.01 in group C,0.23±0.01 and 0.24±0.01 in group D,0.19±0.01 and 0.19±0.01 in group E,and 0.16±0.01 and 0.20±0.02 in group F,both markedly low

关 键 词：糖皮质激素 淫羊藿苷 齐墩果酸 Janus激酶/信号转导及转录因子通路 

分 类 号：R28[医药卫生—中药学]