基于PI3K/Akt/NF-κB途径探究补肾活血方治疗子宫内膜异位症的作用机制研究

Study on the mechanism of Bushen Huoxue Recipe in the treatment of endometriosis based on PI3K/Akt/NF-κB pathway

关键词:  子宫内膜异位症  补肾活血方  PI3K/Akt/NF-κB信号通路  炎症反应  CD31  VEGF  MMP-9

Key words:  ndometriosis  Bushen Huoxue Recipe  PI3K/Akt/NF-κB signaling pathway  inflammatory reaction  CD31  VEGF  MMP-9

摘要:

目的 探究补肾活血方（BSHXR）治疗子宫内膜异位症（EMS）的作用机制。方法 采用自体子宫内膜移植术建立EMS大鼠模型，将大鼠随机分为假手术组、模型组、BSHXR高、中、低剂量组、LY294002（PI3K抑制剂）组，每组10只。给药28 d后，测定大鼠异位子宫内膜病灶体积，苏木精-伊红（HE）染色观察异位子宫内膜组织病理变化，酶联免疫吸附实验（ELISA）法检测血清肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）的水平，免疫组化染色检测异位子宫内膜组织血小板-内皮细胞黏附分子（CD31）表达，实时荧光定量聚合酶链反应（RT-qPCR）检测异位子宫内膜组织血管内皮生长因子（VEGF）、基质金属蛋白酶-9（MMP-9）mRNA水平，Western blot法检测异位内膜组织磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）/核因子κB（NF-κB）通路蛋白表达水平。结果 与假手术组比较，EMS大鼠异位子宫内膜病灶体积显著增加，血清炎症因子水平升高，CD31阳性细胞数量增加，VEGF、MMP-9 mRNA水平升高，PI3K/Akt/NF-κB信号通路被活化（P＜0.05）。与模型组比较，BSHXR可缩小大鼠异位子宫内膜病灶体积，降低血清炎症因子水平，减少CD31阳性细胞数量，降低VEGF、MMP-9 mRNA表达水平，抑制PI3K/Akt/NF-κB信号通路活化，其作用呈剂量依赖性（P＜0.05）。结论 BSHXR对大鼠EMS具有治疗作用，其作用机制与抑制PI3K/Akt/NF-κB信号通路活化有关。

Abstract:

Objective To explore the effect and mechanism of Bushen Huoxue Recipe (BSHXR) on endometriosis (EMS).Methods The EMS rat model was established by autologous endometrial transplantation. The rats were randomly divided into sham operation group, model group, BSHXR high, medium and low dose groups, LY294002 (PI3K inhibitor) group, with 10 rats in each group. After 28 days of administration, the volume of ectopic endometrium was measured. The histopathological changes of the endometrium were observed by HE staining. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) in serum were detected by ELISA. The expression of platelet endothelial cell adhesion molecule-1(CD31) in ectopic endometrium was detected by immunohistochemistry. The mRNA levels of vascular endothelial growth factor(VEGF) and matrix metalloproteinase-9(MMP-9) in ectopic endometrium were detected by RT-qPCR. The protein expression of phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/nuclear factor kappa-B (NF-κB) pathway in ectopic endometrium was detected by Western blot.Results Compared with sham operation group, the volume of ectopic endometrium in model group was significantly increased, the levels of serum inflammatory factors were increased, the number of CD31 positive cells was increased, the mRNA levels of VEGF and MMP-9 were increased, and PI3K/Akt/NF-κB signaling pathway was activated(P < 0.05). Compared with model group, BSHXR could reduce the volume of ectopic endometrium, decrease the serum levels of inflammatory factors, decrease the number of CD31 positive cells, decrease the expression levels of VEGF and MMP-9 mRNA, and inhibit the activation of PI3K/Akt/NF-κB signaling pathway in a dose-dependent manner (P < 0.05).Conclusion BSHXR has therapeutic effects on EMS in rats, and its mechanism may be related to the inhibition of PI3K/Akt/NF-κB signaling pathway activation.