文献类型：期刊文献

中文题名：基于EGFR/PI3K/Akt信号通路探讨加味少腹逐瘀汤治疗子宫内膜异位症的作用机制

英文题名：Mechanism of Modified Shaofu Zhuyutang in Treatment of Endometriosis Based on EGFR/PI3K/Akt Signaling Pathway

作者：杨亚玲[1];王婉润[2];张作良[2];林祥羽[2];王家兴[2];黄灿灿[2];吉秀家[2];武权生[2]

第一作者：杨亚玲

机构：[1]甘肃中医药大学附属医院,兰州730030;[2]甘肃中医药大学中医临床学院,兰州730000

第一机构：甘肃中医药大学第二附属医院

年份：2025

卷号：31

期号：4

起止页码：29

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：;北大核心:【北大核心2023】；

基金：国家自然科学基金项目(82260949);甘肃省自然科学基金项目(23JRRA1199);甘肃省中医药科研项目(GZKP-2022-22);兰州市科技计划项目(2021-1-97,2023-ZD-237,2023-ZD-21)。

语种：中文

中文关键词：子宫内膜异位症;加味少腹逐瘀汤;表皮生长因子受体(EGFR)/磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路;凋亡

外文关键词：endometriosis;modified Shaofu Zhuyutang;epidermal growth factor receptor(EGFR)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)signaling pathway;apoptosis

摘要：目的:观察加味少腹逐瘀汤对子宫内膜异位症SD大鼠表皮生长因子受体(EGFR)/磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路关键蛋白的影响。方法:将60只SPF级雌性SD大鼠按照自体移植法成功复制子宫内膜异位症疾病模型后,随机分为模型组,加味少腹逐瘀汤高、中、低剂量组,孕三烯酮组,另取12只作为空白组。空白组及模型组分别给予10 mL·kg^(-1)生理盐水,加味少腹逐瘀汤高、中、低剂量组分别给予加味少腹逐瘀汤水煎剂30、15、7.5 g·kg^(-1),孕三烯酮组给予孕三烯酮混悬液0.25 mg·kg^(-1)。给药4周后,给予缩宫素2 U诱发宫缩,观察大鼠扭体反应。24 h后处死大鼠,记录各组大鼠异位子宫内膜组织重量、体积。苏木素-伊红(HE)染色观察子宫内膜组织病理变化,原位末端标记法(TUNEL)观察子宫内膜组织凋亡率;免疫荧光法(IF)检测各组大鼠子宫内膜组织中B细胞淋巴瘤-2(Bcl-2)基因相关启动子(Bad)、Bcl-2蛋白表达面积,酶联免疫吸附测定法(ELISA)检测各组大鼠血清中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、表皮生长因子(EGF)、EGFR含量,蛋白免疫印迹法(Western blot)检测各组大鼠子宫内膜组织中EGFR、PI3K、磷酸化磷脂酰肌醇3-激酶(p-PI3K)、Akt、磷酸化蛋白激酶B(p-Akt)蛋白相对表达量,实时荧光定量聚合酶链式反应(Real-time PCR)检测EGFR、PI3K、Akt mRNA表达量。结果:与空白组比较,模型组大鼠病理可见子宫内膜增厚、腺体及间质增生,异位子宫内膜组织Bad相对表达面积显著降低、Bcl-2相对表达面积显著增加、TUNEL染色显示细胞凋亡率显著降低,血清IL-1β、IL-6、TNF-α、EGF、EGFR含量显著升高(P<0.01);EGFR、PI3K、p-PI3K、Akt、p-Akt蛋白相对表达显著上升,EGFR、PI3K、Akt mRNA表达显著上升(P<0.01)。与模型组比较,加味少腹逐瘀汤高、中剂量组及孕三烯酮组腺体和间质不同程度减少,腺腔扩张。扭体次数显著减少、扭体反应潜伏时间显著延长,异位子宫内膜组织体积及质量显著减小,异位子宫内膜组织Bad相对表达面积显著上升、Bcl-2相对表达面积显著下降、TUNEL染色显示细胞凋亡率显著增高,血清IL-1β、IL-6、TNF-α、EGF、EGFR含量显著下降,EGFR、PI3K、p-PI3K、Akt、p-Akt蛋白相对表达显著下降,EGFR、PI3K、Akt mRNA表达明显下降(P<0.05,P<0.01)。结论:加味少腹逐瘀汤可能通过干预EGFR/PI3K/Akt信号通路关键蛋白,诱导异位子宫内膜组织凋亡,对子宫内膜异位症起到治疗作用。
Objective:To observe the effects of modified Shaofu Zhuyutang on key proteins of the epidermal growth factor receptor(EGFR)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)signaling pathway in SD rats with endometriosis.Methods:After successful establishment of an endometriosis model in 60 female SD rats of SPF grade via the auto-transplantation method,the rats were randomly divided into a model group,modified Shaofu Zhuyutang high-,medium-,and low-dose groups,and a gestrinone group,with another 12 rats serving as a blank group.The blank and model groups were administered 10 mL·kg^(-1)normal saline,while the high-,medium-,and low-dose groups received 30,15,and 7.5 g·kg^(-1)modified Shaofu Zhuyutang,respectively.The gestrinone group was administered 0.25 mg·kg^(-1)gestrinone suspension.After four weeks of treatment,uterine contractions were induced with 2 U of oxytocin,and the writhing response of rats was observed.After 24 h,the rats were euthanized,and the weight and volume of ectopic endometrial tissue were recorded.Hematoxylin-eosin(HE)staining was used to observe pathological changes in endometrial tissues,while the terminal deoxynucleotidyl transferase-mediated dUTP nick endlabeling(TUNEL)assay was used to evaluate the apoptosis rate of endometrial tissues.Immunofluorescence was used to detect the relative expression areas of the B-cell lymphoma-2 gene-associated promoter(Bad)and B-cell lymphoma-2(Bcl-2)proteins in endometrial tissues.Serum levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),epidermal growth factor(EGF),and EGFR were measured by enzyme-linked immunosorbent assay(ELISA).The relative protein expression levels of EGFR,PI3K,phosphorylated PI3K(p-PI3K),Akt,and phosphorylated Akt(p-Akt)in endometrial tissues were analyzed by Western blot.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the mRNA expression levels of EGFR,PI3K,and Akt.Results:Compared with the blank group,the model group showed endometrial thickening,glandular and mesenchymal hyperplasia,a significant decrease in the relative expression area of Bad in ectopic endometrial tissues,a significant increase in the relative expression area of Bcl-2,and a significant reduction in the apoptosis rate as indicated by TUNEL staining.Serum levels of IL-1β,IL-6,TNF-α,EGF,and EGFR were significantly elevated(P<0.01).The relative protein expression levels of EGFR,PI3K,p-PI3K,Akt,and p-Akt,as well as the mRNA expression levels of EGFR,PI3K,and Akt,were also significantly increased(P<0.01).Compared with the model group,the high-and mediumdose groups of modified Shaofu Zhuyutang and the gestrinone group exhibited reduced glandular and mesenchymal hyperplasia to varying degrees,with dilated glandular lumens.The number of writhing responses was significantly reduced,the latency to writhing response was significantly prolonged,and the weight and volume of ectopic endometrial tissue were significantly decreased.The relative expression area of Bad in ectopic endometrial tissue was significantly increased,the relative expression area of Bcl-2 was significantly decreased,and the apoptosis rate was significantly elevated as shown by TUNEL staining.Serum levels of IL-1β,IL-6,TNF-α,EGF,and EGFR were significantly reduced,and the relative protein expression levels of EGFR,PI3K,p-PI3K,Akt,and p-Akt,as well as the mRNA expression levels of EGFR,PI3K,and Akt,were significantly decreased(P<0.05,P<0.01).Conclusion:Modified Shaofu Zhuyutang may exert therapeutic effects on endometriosis by interfering with key proteins of the EGFR/PI3K/Akt signaling pathway and inducing apoptosis in ectopic endometrial tissue.