摘要
目的 探讨补肾养营活血方(BYHF)对大鼠肾虚血瘀型宫腔粘连的预防作用及机制。方法 将24只雌性SD大鼠随机分为对照组(Control)、模型组(Model)、补肾养营活血方组(BYHF),每组8只。采用宫腔机械搔刮损伤法结合肾上腺素皮下注射、羟基脲溶液灌胃制备肾虚血瘀型IUA病证结合动物模型。BYHF组予补肾养营活血方(2.943g·kg^(-1)·d^(-1))灌胃,对照组、模型组灌服等体积蒸馏水,连续3周。动情期麻醉后取材,ELISA检测肾虚(cAMP、cAMP/cGMP)、血瘀(ET、TXB2)证候指标,HE和Masson染色分别观察子宫内膜厚度、腺体数及纤维化程度,Western blot(WB)检测子宫内膜组织NLRP3、Caspase-1、IL-18蛋白表达。体外LPS与Nigericin诱导HESCs焦亡模型,予BYHF含药血清培养,Calcein AM/PI染色观察细胞焦亡,ELISA检测细胞上清Caspase-1、IL-1β和IL-18水平。结果 补肾养营活血方可以改善肾虚血瘀证候(P<0.01,P<0.05),减轻IUA大鼠子宫内膜纤维化程度(P<0.01),改善子宫内膜厚度和腺体数(P<0.05),抑制NLRP3(P<0.01)、IL-18(P<0.05)、Caspase-1(P<0.01)蛋白表达,其含药血清可抑制HESCs焦亡,减少Caspase-1、IL-1β和IL-18等细胞焦亡因子的表达(P<0.05)。结论 补肾养营活血方通过改善肾虚血瘀IUA模型大鼠肾虚血瘀证候,抑制细胞焦亡等方面来预防宫腔粘连、促进内膜再生。
Objective To explore the intervening effect and mechanism of Bushen Yangying Huoxue Formula(补肾养营活血方,BYHF)on intrauterine adhesions(IUA)rats with kidney deficiency and blood stasis.Methods A total of 24 adult female Sprague-Dawley(SD)rats were randomized into the Control group,Model group,and BYHF group,with 8 rats each.A rat model of IUA with kidney deficiency and blood stasis was established through mechanical damage,subcutaneous injection of epinephrine,and intragastric administration of hydroxyurea solution.The rats in the BYHF group were administered BYHF intragastrically(2.943g kg^(-1)·d^(-1))for 3 weeks and those from the Control and Model groups were administered the same volume of distilled water.Tissue samples were collected from anesthetized rats in estrous cycles.The syndrome of kidney deficiency(evaluated by cAMP,cAMP/cGMP)and blood stasis(evaluated by ET,TXB2)were assessed by ELISA.The thickness of the endometrium,number of glands and fibrosis rate were evaluated using HE and Masson staining,respectively.Western blot(WB)was utilized to detect the protein levels of NLRP3,Caspase-1,and IL-18.Human endometrial stromal cells(HESCs)were induced by LPS and Nigericin to establish in vitro pyroptosis models.BYHFcontaining serum was used to culture HESCs.Pyroptotic cells were visualized via staining with Calcein AM/PI.The concentrations of Caspase-1,IL-1β and IL-18 in the supernatants were assessed by ELISA.Results BYHF was able to alleviate the syndrome of kidney deficiency and blood stasis of IUA(P<0.01,P<0.05),reduce the degree of fibrosis(P<0.01),improve the thickness of the endometrium and the number of glands(P<0.05),and downregulate the expressions of NLRP3(P<0.01),Caspase-1(P<0.01),and IL-18(P<0.05).BYHF-containing serum inhibited pyroptosis of HESCs and the expression of the pyroptosis-related factors(Caspase-1,IL-1βand IL-18)(P<0.05).Conclusion BYHF could alleviate the syndrome of kidney deficiency and blood stasis,and inhibit pyroptosis for preventing IUA and promoting endometrial regeneration.
作者
陈琇涵
杨茜
李方远
CHEN Xiuhan;YANG Qian;LI Fangyuan(Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China;West China Second University Hospital,Chengdu 610041,China)
出处
《时珍国医国药》
北大核心
2025年第20期3846-3852,共7页
JOURNAL OF LI-SHIZHEN TRADITIONAL CHINESE MEDICINE
基金
中国博士后科学基金面上资助项目(2022M722295)
国家自然科学基金青年基金(82305302)
四川省重点研发项目(2024YFFK0167)。
