补肾活血方对子宫内膜氧化应激及纤维化调控机制的研究  

Mechanism of Bushen Huoxue Formula in regulating endometrial oxidative stress and fibrosis

作　　者：汤锶锶 陈蓉[1] 宋谦谦 TANG Si-si;CHEN Rong;SONG Qian-qian(College of Traditional Chinese Medicine,Chongqing Medical University,Chongqing 400016,China;Chongqing Hospital of Traditional Chinese Medicine,Chongqing 400021,China)

机构地区：[1]重庆医科大学中医药学院,重庆400016 [2]重庆市中医院,重庆400021

出　　处：《中国中药杂志》2025年第1期218-227,共10页China Journal of Chinese Materia Medica

基　　金：重庆市科研机构绩效激励引导专项(JXYN2020-3,cstc2021jxjl130023)。

摘　　要：宫腔粘连(intrauterine adhesion,IUA)是常见难治性妇科疾病,缺乏特异性有效药物及措施防治子宫内膜纤维化。该研究通过补肾活血方干预机械损伤联合感染双重介导法构建的大鼠IUA模型对子宫内膜氧化应激及纤维化调控的机制进行探讨。将72只8~10周龄SPF级SD雌性大鼠随机分为空白对照组、模型对照组、补肾活血方低剂量组、补肾活血方中剂量组、补肾活血方高剂量组、雌激素组,对监测到处于动情周期的模型对照组及阳性对照组大鼠进行手术损伤+感染造模,假手术组仅做开关腹处理。造模成功后,模型对照组每天灌胃纯净水15μL·g^(-1);低、中、高剂量组灌胃补肾活血方7.8、15.6、31.2 g·kg^(-1),雌激素组灌胃戊酸雌二醇4.2 mg·kg^(-1)。连续干预28 d后禁食不禁水,全部处死后取血与组织待用。苏木精-伊红(hematoxylin-eosin,HE)染色计算子宫腺体数目,马松(Masson)染色计算子宫胶原纤维面积,并结合HE、Masson染色对子宫内膜纤维化程度进行半定量评分;免疫组化检测大鼠子宫组织,血管内皮生长因子(vascular endothelial growth factor,VEGF)、基质细胞衍生因子-1(stromal cell-derived factor-1,SDF-1)及转录生长因子-β1(transforming growth factor-β1,TGF-β1)表达情况;酶联免疫吸附测定(ELISA)法检测大鼠血清中干扰素-γ(interferon-γ,IFN-γ)、白细胞介素(interleukin,IL)-1α、TGF-β1、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、Ⅳ型胶原(collagen typeⅣ,Ⅳ-Col)、白血病抑制因子(leukemia inhibitory factor,LIF)、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px);qPCR测定Smad2、Smad3、解整合素金素蛋白酶(adisintegrin and metalloproteinase,ADAM17)、TGF-β1 mRNA表达;Western blot检测大鼠子宫Notch和ADAM17的蛋白表达。结果显示,补肾活血方高剂量干预后子宫纤维化面积显著降低,水肿、粘连情况有效缓解;炎症因Intrauterine adhesion(IUA)is a common gynecological disease that is difficult to treat,and there is a lack of specific effective drugs and measures to prevent endometrial fibrosis.In this study,the mechanism of endometrial oxidative stress and fibrosis regulation was studied in an IUA rat model constructed by Bushen Huoxue Formula intervention of mechanical injury combined with infection.A total of 72 SPF SD female rats aged 8-10 weeks were randomly divided into blank control group,model control group,low-dose,medium-dose,and high-dose groups of Bushen Huoxue Formula,and estrogen groups.The rats in the estrous cycle of the model control group and the positive control group were simulated with surgical injury and infection,and the sham operation group was treated with on-off abdominal treatment.After successful modeling,the model control group was administered intragastrically with purified water of 15μL·g^(-1)every day.The low-dose group was administered intragastrically with Bushen Huoxue Formula of 7.8 g·kg^(-1);the medium-dose group was administered intragastrically with Bushen Huoxue Formula of 15.6 g·kg^(-1),and the high-dose group was administered intragastrically with Bushen Huoxue Formula of 31.2 g·kg^(-1).The estrogen group was administered intragastrically with estradiol valerate of 4.2 mg·kg^(-1).After continuous intervention for 28 days,all rats were deprived of water and killed to collect blood and tissue.Hematoxylin-eosin(HE)staining calculated the number of uterine glands;Masson staining calculated the area of uterine collagen fibers.Combined with HE and Masson staining,semi-quantitative scores were performed on the degree of endometrial fibrosis.Immunohistochemistry was performed to detect the vascular endothelial growth factor(VEGF),stromal cell-derived factor-1(SDF-1),and transforming growth factor-β1(TGF-β1)expression in rats′uterine tissue.Enzyme-linked immunosorbent assay(ELISA)dected angiopoietin 1(IFN-γ),interleukin-1α(IL-1α),TGF-β1,tumor necrosis factor-α(TNF-α),collagen ty

关 键 词：宫腔粘连 补肾活血方 TGF-β1/Smads信号通路 ADAM17/Notch信号通路 

分 类 号：R285.5[医药卫生—中药学]