文献类型：期刊文献

中文题名：加味少腹逐瘀汤介导VEGF/PI3K/Akt/eNOS信号通路抑制子宫内膜异位症血管生成的作用机制

英文题名：Modified Shaofu Zhuyutang Mediates VEGF/PI3K/Akt/eNOS Signaling Pathway to Inhibit Angiogenesis in Endometriosis

作者：王家兴[1];史奇[3];武权生[1,2]

第一作者：王家兴

机构：[1]甘肃中医药大学中医临床学院,兰州730000;[2]甘肃中医药大学附属医院,兰州730030;[3]甘肃中医药大学药学院,兰州730000

第一机构：甘肃中医药大学中医临床学院

年份：2025

卷号：31

期号：7

起止页码：81

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)。

语种：中文

中文关键词：加味少腹逐瘀汤;子宫内膜异位症;血管生成;血管内皮生长因子(VEGF)/磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)/内皮型一氧化氮合酶(eNOS);一氧化氮(NO)

外文关键词：modified Shaofu Zhuyutang;endometriosis;angiogenesis;vascular endothelial growth factor(VEGF)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/endothelial nitric oxide synthase(eNOS);nitric oxide(NO)

摘要：目的:探讨加味少腹逐瘀汤介导血管内皮生长因子(VEGF)/磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)/内皮型一氧化氮合酶(eNOS)-一氧化氮(NO)信号通路抑制子宫内膜异位症血管生成的作用机制。方法:84只SD雌性大鼠随机分为正常组,假手术组,模型组,孕三烯酮组,加味少腹逐瘀汤(中药)高、中、低剂量组,采用自体移植法进行大鼠子宫内膜异位症模型建造,造模成功后中药高、中、低剂量组分别给予加味少腹逐瘀汤浓缩液30、15、7.5 g·kg^(-1)灌胃给药,孕三烯酮组给予孕三烯酮混悬液0.25 mg·kg^(-1)灌胃给药,其余正常组、假手术组、模型组给予等量蒸馏水。灌胃28 d后给予缩宫素观察大鼠扭体反应次数及潜伏时间,收集各组大鼠血清、各造模组大鼠异位灶及正常组与假手术组大鼠子宫,苏木素-伊红(HE)染色观察大鼠子宫内膜异位灶病理形态学变化,免疫组化法观察血管生成特异性指标簇分化34抗原(CD34)、FLI-1转录因子(FLI-1)表达情况,酶联免疫吸附测定法(ELISA)检测血清VEGF水平,硝酸还原酶法检测血清NO含量,蛋白免疫印迹法(Western blot)检测组织VEGF、PI3K、磷酸化的磷脂酰肌醇3-激酶(p-PI3K)、Akt、磷酸化的蛋白激酶B(p-Akt)、eNOS、磷酸化的内皮型一氧化氮合酶(p-eNOS)蛋白水平,实时荧光定量聚合酶链式反应(Real-time PCR)检测VEGF、PI3K、Akt、eNOS mRNA水平。结果:与正常组比较,假手术组大鼠扭体反应次数及扭体反应时间、血清VEGF及NO表达水平、组织VEGF蛋白、p-PI3K/PI3K、p-Akt/Akt、p-eNOS/eNOS及VEGF、PI3K、Akt、eNOS mRNA水平变化均差异无统计学意义;模型组大鼠扭体反应次数显著增加,扭体反应潜伏时间显著缩短,腹壁可见异位子宫内膜组织显著增大,出现间质增生、腺体扩张和血管增多,CD34、FIL-1阳性表达率显著升高,血清VEGF及NO表达水平、蛋白VEGF、p-PI3K/PI3K、p-Akt/Akt、p-eNOS/eNOS及VEGF、PI3K、Akt、eNOS mRNA表达水平显著上调(P<0.01);与模型组比较,孕三烯酮组和中药高、中、低剂量组扭体反应次数明显减少,扭体反应时间明显延长,腹壁异位子宫内膜组织明显减小,病理损伤程度较模型组明显减轻,CD34、FIL-1阳性表达率明显降低,孕三烯酮组和中药高、中剂量组血清VEGF及NO表达水平、蛋白VEGF、p-PI3K/PI3K、p-Akt/Akt、p-eNOS/eNOS及VEGF、PI3K、Akt、eNOS mRNA表达水平明显下调;中药低剂量组血清VEGF,蛋白VEGF、p-PI3K/PI3K、p-Akt/Akt、p-eNOS/eNOS及VEGF、eNOS mRNA表达水平明显降低(P<0.05,P<0.01)。结论:加味少腹逐瘀汤可通过拮抗VEGF/PI3K/Akt/eNOS-NO信号通路异常激活,抑制子宫内膜异位症血管生成,从而阻止子宫内膜异位症的发生发展及恶化。
Objective:To explore the mechanism by which modified Shaofu Zhuyutang inhibits angiogenesis in endometriosis via the vascular endothelial growth factor(VEGF)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/endothelial nitric oxide synthase(eNOS)-nitric oxide(NO)signaling pathway.Methods:Eighty-four female SD rats were randomly assigned into blank,sham operation,model,positive control(gestrinone,0.25 mg·kg^(-1)),high-,medium-,and lowdose(30,15,7.5 g·kg^(-1),respectively)traditional Chinese medicine(TCM,modified Shaofu Zhuyutang)groups.A rat model of endometriosis was established by the autotransplantation method.After successful modeling,rats in the drug intervention groups were administrated with corresponding agents by gavage,and those in the blank,sham operation,and model groups received an equal volume of distilled water.After 28 days of gavage,rats were administrated with oxytocin,and the number and latency period of writhing responses were observed.Serum samples from each group,ectopic lesions from modeling groups,and uteri from blank and sham operation groups were collected.Hematoxylin-eosin staining was used to observe the pathological morphology of endometriotic lesions.Immunohistochemistry was employed to observe the expression of angiogenesis-specific markers cluster of differentiation 34 antigen(CD34)and friend leukemia virus integration-1(FLI-1).Enzyme-linked immunosorbent assay and the nitrate reductase method were employed to determine the serum levels of VEGF and NO,respectively.Western blot was employed to measure the protein levels of VEGF,PI3K,phosphorylated PI3K(p-PI3K),Akt,phosphorylated Akt(p-Akt),eNOS,and phosphorylated eNOS(p-eNOS).Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was conducted to determine the mRNA levels of VEGF,PI3K,Akt,and eNOS.Results:The blank group and the sham operation group had no significant changes in the number and latency period of writhing responses,serum VEGF and NO levels,protein levels of VEGF,p-PI3K/PI3K,p-Akt/Akt,and p-eNOS/eNOS,and mRNA levels of VEGF,PI3K,Akt,and eNOS.The model group showed an increase in the number and a reduction in the latency period of writhing responses,enlargement of ectopic endometrial tissue in the abdominal wall,with stromal hyperplasia,glandular dilation,and increased vasculature.In addition,the modeling led to increased positive expression of CD34 and FLI-1,elevated serum VEGF and NO levels,and up-regulated protein levels of VEGF,p-PI3K/PI3K,p-Akt/Akt,and p-eNOS/eNOS and mRNA levels of VEGF,PI3K,Akt,and eNOS(P<0.01).Compared with the model group,the gestrinone and high-,medium-,and low-dose TCM groups showed a significant reduction in the number of writhing responses,a significant prolongation of the latency period,reduced ectopic endometrial tissue in the abdominal wall,alleviated pathological damage,and reduced positive expression of CD34 and FLI-1.The gestrinone group and the high-and medium-dose TCM groups showed lowered serum VEGF and NO levels as well as down-regulated protein levels of VEGF,p-PI3K/PI3K,p-Akt/Akt,and p-eNOS/eNOS and mRNA levels of VEGF,PI3K,Akt,and eNOS.Moreover,the low-dose TCM group showed reductions in the serum VEGF level,the protein levels of VEGF,p-PI3K/PI3K,p-Akt/Akt,and p-eNOS/eNOS,and the mRNA levels of VEGF and eNOS(P<0.05,P<0.01).Conclusion:Modified SShaofu Zhuyutang can inhibit angiogenesis in endometriosis by antagonizing the abnormal activation of the VEGF/PI3K/Akt/eNOS-NO signaling pathway,thereby preventing the occurrence,development,and deterioration of endometriosis.