摘要
目的探索实验性自身免疫性甲状腺炎(EAT)诱导的小鼠卵巢储备功能下降(DOR)模型,并探讨接绪资冲颗粒(ZCWS)的保护机制。方法将96只昆明雌性小鼠随机分为J对照组、BIW对照组、J模型组和BIW模型组,通过抗原免疫(0.1 mg·1次·周^(-1)或2次·周^(-1))联合高碘水(0.64 g·L^(-1))喂养7-19周,验证模型。然后将30只小鼠随机分为ZCWS组、模型组和对照组,抗原免疫(0.2 mg·周^(-1))+高碘水喂养13周建立EAT诱导的DOR模型。ZCWS组灌胃中药混悬液(0.64 g·mL^(-1)),其余组灌胃等体积生理盐水,持续7周。通过HE染色、ELISA、IHC和Western blot检测甲状腺和卵巢组织形态及氧化应激相关指标。结果抗原免疫13周(0.1-0.2 mg·周^(-1))配合高碘水喂养可较稳定地构建EAT的DOR模型。ZCWS减轻甲状腺淋巴细胞浸润程度、滤泡结构破坏,血清TPOAb和TGAb水平(P<0.01);抑制MDA活力(P<0.01),增加GSH-Px和SOD活力(P<0.05);增加原始、初级和次级卵泡(P<0.05),减少闭锁卵泡(P<0.01);上调AMH和下调FSH(P<0.01);调节卵巢组织Bcl-2、Bax和Caspase-3蛋白表达;下调Keap1蛋白(P<0.01),上调Nrf2(P<0.01)、HO-1和LONP1蛋白(P<0.05)。上调卵巢组织Bcl-2蛋白(P<0.05),下调Bax(P<0.01)和Caspase-3(P<0.05)蛋白;下调Keap1蛋白(P<0.01),上调Nrf2蛋白(P<0.01)和HO-1、LONP1蛋白(P<0.05)。结论ZCWS通过激活Nrf2通路抑制氧化应激,减少卵泡闭锁,改善EAT小鼠的卵巢储备功能。
Objective To explore the model of decreased ovarian reserve(DOR)induced by experimental autoimmune thyroiditis(EAT)in mice and investigate the protective mechanism of Jiexu Zichong Granules(ZCWS).Methods Ninety-six Kunming female mice were randomly divided into the J control group,BIW control group,J model group,and BIW model group.The mice were immunized with antigen(0.1 mg,once or twice per week)combined with high iodine water(0.64 g/L)for 7-19 weeks to validate the model.Subsequently,30 mice were randomly divided into the ZCWS group,model group,and control group.The DOR model was established by immunization with the antigen(0.2 mg per week)combined with high iodine water feeding for 13 weeks.The ZCWS group received oral administration of traditional Chinese medicine suspension(0.64 g/mL),while the remaining groups received an equivalent volume of physiological saline for 7 weeks.Thyroid and ovarian tissue morphology and related indicators were detected using HE staining,ELISA,IHC,and Western blot.Results Antigen immunization for 13 weeks(0.1-0.2 mg per week)combined with high iodine water feeding stably constructed the EAT DOR model.ZCWS reduced thyroid lymphocyte infiltration,follicular structure destruction,and serum TPOAb and TGAb levels(P<0.01).It inhibited MDA activity(P<0.01),increased GSHPx and SOD activities(P<0.05),increased primordial,primary,and secondary follicles(P<0.05),and reduced atretic follicles(P<0.01).ZCWS upregulated AMH and downregulated FSH(P<0.01),regulated the expression of Bcl-2,Bax,and Caspase-3 proteins in ovarian tissue,downregulated Keap1 protein(P<0.01),and upregulated Nrf2(P<0.01),HO-1,and LONP1 proteins(P<0.05).It upregulated Bcl-2 protein(P<0.05),downregulated Bax(P<0.01),and Caspase-3(P<0.05)proteins,downregulated Keap1 protein(P<0.01),and upregulated Nrf2(P<0.01),HO-1,and LONP1 proteins(P<0.05)in ovarian tissue.Conclusion ZCWS improves ovarian reserve function in EAT mice by activating the Nrf2pathway,inhibiting oxidative stress,and reducing follicular atresia.
作者
李方远
陆华
张琦
杨茜
强玲侠
刘芊辰
LI Fangyuan;LU Hua;ZHANG Qi;YANG Qian;QIANG Lingxia;LIU Qianchen(Chengdu University of Traditional Chinese Medicine,Chengdu 610032,China;Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610032,China)
出处
《世界科学技术-中医药现代化》
北大核心
2025年第2期581-591,共11页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
四川省中医药科技产业创新团队专项(2022C003):中医药调控女性生殖健康产品开发创新团队,负责人:陆华
国家重点研发计划中医药现代化研究不孕症“肾阳虚证”辨证标准的系统研究(2018YFC1704305),负责人:陆华
四川省中医药院士后备人才培养项目(川中医药函[2020]73号),负责人:陆华。
关键词
抗原免疫
自身免疫性甲状腺炎
接绪资冲颗粒
卵巢储备
氧化应激
抗凋亡
Antigen immunization
Autoimmune thyroiditis
Jiexu Zichong granules
Ovarian reserve
Oxidative stress
Anti-apoptotic
作者简介
通讯作者:陆华(ORCID:0000-0002-2709-222X),研究员,主要研究方向:中医药对女性生殖调控的研究。
