益气消癥法方含药血清干预EA.hy926细胞MAPK信号转导通路ERK1/2表达的研究  

Intervention of Yiqi Xiaozheng Prescription-contained serum on expression of ERK1/2 based on MAPK signal pathway in EA.hy 926 cells

作　　者：李爽[1] 刘丽丽[2] 夏天[3] 邓高丕[4] 宋卓敏[5] LI Shuang1 LIU Li-li2 XIA Tian3 DENG Gao-pi4 SONG Zhou-min5 1. Guangzhou University of Chinese Medicine Guangzhou 510504 China 2. Yuebei People＇s Hospital Shaoguan 512026 China 3. First Teaching Hospital of Tianjin University ofTCM Tianjin 300193 China 4. First Affiliated Hospital of Guangzhou University of Chinese Medicine Guangzhou 510405 China 5. Research Institute ofTCM Tianjin University ofTCM

机构地区：[1]广州中医药大学,广东广州510504 [2]粤北人民医院,广东韶关512026 [3]天津中医药大学第一附属医院,天津300193 [4]广州中医药大学第一附属医院,广东广州510504 [5]天津中医药大学中医药研究院,天津300193

出　　处：《药物评价研究》2016年第5期766-770,共5页Drug Evaluation Research

基　　金：国家自然科学基金面上项目(30973768)

摘　　要：目的研究益气消癥法方含药血清对EA.hy926细胞丝裂原活化蛋白激酶(MAPK)信号通路上细胞外信号调节激酶1/2(ERK1/2)表达的影响,探索其抑制子宫肌瘤血管新生的作用机制。方法 SD大鼠随机分为:对照组,益气消癥法方高、中、低剂量(生药剂量24、12、6 g/kg)组,ig给药,每天给药1次,连续给药5 d,腹主动脉取血,分离血清,灭活、过滤除菌;体外培养EA.hy926细胞,随机分为6组:对照血清组、模型组及益气消癥法方高、中、低剂量血清组和氟维司群(ICI,阳性药)组,除对照血清组外,其余组别给予雌二醇(E2),诱导细胞增殖,制备子宫肌瘤血管模型;通过实时荧光定量PCR(q RT-PCR)及Western blotting法检测MEK2和ERK1/2基因与蛋白的表达。结果模型组MEK2和ERK1/2基因与蛋白的表达水平明显高于对照血清组(P<0.01);与模型组比较,ICI和益气消癥法方高剂量血清组MEK2和ERK1/2表达水平显著降低(P<0.01)。结论益气消癥法方含药血清通过降调ERK1/2表达,阻断MAPK/ERK1/2信号转导,抑制雌激素诱导的血管内皮细胞增殖,减少子宫肌瘤血液供应。Objective To investigate the intervention of Yiqi Xiaozheng Prescription（YXP）-contained serum on expression of extracellular-signal regulated kinase 1/2（ERK1/2） based on mitogen-activated protein kinase（MAPK） signal pathway in EA.hy926 cells, and to investigate the mechanism of inhibiting angiogenesis in uterine leiomyoma. Methods SD rats were divided into four groups： control group; YXP low, medium, and high dose groups（24, 12, and 6 g/kg at crude drug dosage）. After sc administration once daily for 5 d, blood plasma was collected by abdominal aortic method, serum was isolated, inactivated, and filtrated rid of bacterium. EA.hy926 cells cultured in vitro were divided into six groups： control serum group; model group, YXP low, medium, and high dose contained serum group, and filtration（ICI, positive drug） group. Proliferation of EA.hy926 cells was induced by 17β-estradiol（E2） to simulate angiogenesis in uterine leiomyoma except control serum group. Real-time fluorescence quantification PCR（q RT-PCR） and Western blotting were used to detect the gene and protein expression of MEK2 and ERK1/2. Results MEK2 and ERK1/2 gene and protein expression levels of model group were significantly higher than those of control group（P 0.01）; Compared with model group, MEK2 and ERK1/2 expression levels of YXP high dose contained serum group and ICI group were significantly decreased（P 0.01）. Conclusion YXP-contained serum could down-regulate the expression of ERK1/2, block the MAPK/ERK1/2 signal transduction, and inhibit the proliferation of vascular endothelial cells which was induced by estrogen in order to control angiogenesis and reduce blood supply of uterine leiomyoma.

关 键 词：益气消癥法方含药血清 子宫肌瘤 EA.hy926细胞 MAPK信号转导通路 ERK1/2 

分 类 号：R965[医药卫生—药理学]