目的 研究益氣消癥法方含藥血清對EA.hy926細(xì)胞絲裂原活化蛋白激酶（MAPK）信號通路上細(xì)胞外信號調(diào)節(jié)激酶1/2（ERK1/2）表達(dá)的影響，探索其抑制子宮肌瘤血管新生的作用機(jī)制。方法 SD大鼠隨機(jī)分為：對照組，益氣消癥法方高、中、低劑量（生藥劑量24、12、6 g/kg）組，ig給藥，每天給藥1次，連續(xù)給藥5 d，腹主動脈取血，分離血清，滅活、過濾除菌；體外培養(yǎng)EA.hy926細(xì)胞，隨機(jī)分為6組：對照血清組、模型組及益氣消癥法方高、中、低劑量血清組和氟維司群（ICI，陽性藥）組，除對照血清組外，其余組別給予雌二醇（E₂），誘導(dǎo)細(xì)胞增殖，制備子宮肌瘤血管模型；通過實(shí)時熒光定量PCR（qRT-PCR）及Western blotting法檢測MEK2和ERK1/2基因與蛋白的表達(dá)。結(jié)果 模型組MEK2和ERK1/2基因與蛋白的表達(dá)水平明顯高于對照血清組（P<0.01）；與模型組比較，ICI和益氣消癥法方高劑量血清組MEK2和ERK1/2表達(dá)水平顯著降低（P<0.01）。結(jié)論 益氣消癥法方含藥血清通過降調(diào)ERK1/2表達(dá)，阻斷MAPK/ERK1/2信號轉(zhuǎn)導(dǎo)，抑制雌激素誘導(dǎo)的血管內(nèi)皮細(xì)胞增殖，減少子宮肌瘤血液供應(yīng)。

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 (qRT-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.

國家自然科學(xué)基金面上項(xiàng)目（30973768）