银杏二萜内酯通过调控TLR4/MyD88/NF-κB信号通路改善多囊卵巢综合征大鼠卵巢功能OA
Effect of Ginkgo Diterpene Lactones on Improving Ovarian Function in Rats with Polycystic Ovary Syndrome by Regulating TLR4/MyD88/NF-κB Signaling Pathway
目的:探讨银杏二萜内酯对多囊卵巢综合征(polycystic ovary syndrome,PCOS)大鼠卵巢功能的影响及潜在机制.方法:取 48 只 SPF 级雌性 SD 大鼠按随机数字表法分为正常对照组、模型对照组、二甲双胍100 mg/kg 组(灌胃给药)、银杏二萜内酯1.2、2.4、4.8 mg/kg 组(腹腔注射给药),每组 8 只.造模大鼠以来曲唑1 mg/kg 灌胃联合高脂饲料喂养28 d 的方法复制 PCOS 大鼠模型.造模完成后,各组大鼠分别给予相应药物或生理盐水,1 次/d,连续给药 14 d.测定空腹血糖(FBG)、空腹胰岛素(FINS)并计算胰岛素抵抗指数(HOMA-IR);称体质量和卵巢质量,计算卵巢指数;ELISA 法测定血清性激素[雌二醇(E2)、睾酮(T)、促性腺激素释放激素(GnRH)、促卵泡生成素(FSH)、促黄体生成素(LH)]含量和卵巢组织中炎症因子[肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-6]含量;苏木素-伊红(HE)染色法观察卵巢病理学改变,计数囊性卵泡和黄体数;RT-PCR 法测定卵巢组织中 Toll 样受体4(Tlr4)、髓样分化因子 88(Myd88)、核因子-κB p65(Nfκb p65)mRNA 表达;Western blot 法测定 TLR4、MYD88、胞浆 NF-κB p65、胞核 NF-κB p65 蛋白表达.结果:与正常对照组相比,模型对照组大鼠 FBG、FINS 含量、HOMA-IR、体质量、卵巢指数显著升高(P<0.01),血清 T 含量、GnRH、LH 活力和 LH/FSH 比值显著升高,E2 含量、FSH 活力显著降低(P<0.01),卵巢组织中 TNF-α、IL-1β、IL-6 含量显著升高(P<0.01),卵巢组织呈现多囊性扩张,大卵泡增多、成熟卵泡减少、卵泡颗粒层排列紊乱且数目减少、黄体数目减少等病变,囊性卵泡数目显著升高、黄体数目显著降低(P<0.01),卵巢组织中 Tlr4、Myd88、Nfκb p65 mRNA 表达和TLR4、MYD88、胞浆 NF-κB p65、胞核 NF-κB p65 蛋白表达显著上调(P<0.01);与模型对照组相比,二甲双胍100 mg/kg 组和银杏二萜内酯2.4、4.8 mg/kg 组大鼠 FBG、FINS 含量、HOMA-IR、体质量、卵巢指数明显降低(P<0.05 或 P<0.01),血清 T 含量、GnRH、LH 活力和 LH/FSH 比值明显降低,E2 含量、FSH 活力明显升高(P<0.05或 P<0.01),卵巢组织中 TNF-α、IL-1β、IL-6 含量明显降低(P<0.05 或 P<0.01),卵巢组织病变明显改善,囊性卵泡数目显著降低、黄体数目显著升高(P<0.01),卵巢组织 Tlr4、Myd88、Nfκb p65 mRNA 表达和 TLR4、MYD88、胞浆 NF-κB p65、胞核 NF-κB p65 蛋白表达明显下调(P<0.05 或 P<0.01).结论:银杏二萜内酯能够通过调节内分泌水平,抑制炎症,从而有效改善 PCOS 大鼠卵巢功能,银杏二萜内酯上述作用具有一定的剂量依赖性,其作用机制可能与抑制 TLR4/MyD88/NF-κB 信号通路有关.
Objective:To investigate the effect and potential mechanism of ginkgo diterpene lactones on ovarian function in rats with polycystic ovary syndrome(PCOS).Methods:According to the random number table method,48 specific-pathogen free(SPF)female Sprague Dawley(SD)rats were randomly divided into a normal control group,a model control group,a metformin group of 100 mg/kg(intragastric administration),ginkgo diterpene lactone groups of 1.2,2.4,and 4.8 mg/kg(intraperitoneal injection),with eight rats in each group.Modeling rats were intragastrically administered with 1 mg/kg letrozole combined with a high-fat diet for 28 days to establish a PCOS rat model.After mod-eling,rats in each group were given corresponding drugs or saline water,once a day for 14 days.The fasting blood glu-cose(FBG)and fasting insulin(FINS)were measured,and the homeostatic model assessment for insulin resistance(HOMA-IR)was calculated.The body mass and ovarian mass were weighed,and the ovarian index was calculated.The level of sex hormones[estradiol(E2),testosterone(T),gonadotropin-releasing hormone(GnRH),follicle stimulating hormone(FSH),and luteinizing hormone(LH)]in serum and the level of inflammatory factors[tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and IL-6]in ovarian tissue were detected by enzyme-linked immunosorbent assay(ELISA).The pathological changes in the ovary were examined by hematoxylin eosin(HE)staining,and the number of cystic follicles and corpus luteum was counted.The mRNA expression of toll-like receptor 4(Tlr4),myeloid differentia-tion factor 88(Myd88),and nuclear factor kappa B p65(Nfκb p65)in ovarian tissue was detected by reverse transcrip-tion polymerase chain reaction(RT-PCR).The protein expression of TLR4,MYD88,cytoplasmic NF-κB p65,and nucle-ar NF-κB p65 was detected by Western blot.Results:Compared with those in the normal control group,the FBG,FINS,HOMA-IR,body mass,and ovarian index of the rats were significantly increased in the model control group(P<0.01).The levels of T,GnRH,and LH in serum,and LH/FSH were significantly increased,while the level of E2 and FSH was significantly decreased(P<0.01).The level of TNF-α,IL-1β,and IL-6 in ovarian tissue was significantly increased(P<0.01).In ovarian tissue,polycystic dilation,increased large follicles,decreased mature follicles,disordered arrange-ment and decreased number of follicular granulosa layer,and decreased number of corpus luteum were observed.The number of cystic follicles was significantly increased,while the number of corpus luteum was significantly decreased(P<0.01).The mRNA expression of Tlr4,Myd88,and Nfκb p65 and the protein expression of TLR4,MYD88,cytoplasmic NF-κB p65,and nuclear NF-κB p65 in ovarian tissue were significantly upregulated(P<0.01).Compared with those in the model control group,the FBG,FINS,HOMA-IR,body mass,and ovarian index of the rats in the metformin group of 100 mg/kg and the ginkgo diterpene lactones groups of 2.4 and 4.8 mg/kg were significantly decreased(P<0.05 or P<0.01).The levels of T,GnRH,and LH in serum,and LH/FSH were significantly decreased,and the level of E2 and FSH was significantly increased(P<0.05 or P<0.01).The level of TNF-α,IL-1β,and IL-6 in ovarian tissue was sig-nificantly decreased(P<0.05 or P<0.01).The pathological changes in ovarian tissue were significantly improved.The number of cystic follicles was significantly decreased,while the number of corpus luteum was significantly increased(P<0.01).The mRNA expression of Tlr4,Myd88,and Nfκb p65 and the protein expression of TLR4,MYD88,cytoplasmic NF-κB p65,and nuclear NF-κB p65 in ovarian tissue were significantly downregulated(P<0.05 or P<0.01).The effects of the ginkgo diterpene lactone groups of 1.2,2.4,and 4.8 mg/kg on various detection indices(excluding the body mass,GnRH,and IL-6)showed a dose-dependent way(P<0.05 or P<0.01).Conclusion:Ginkgo diterpene lac-tones can effectively improve the ovarian function of PCOS rats by regulating the level of endocrine and inhibiting inflam-mation.The above effects of ginkgo diterpene lactones are dose-dependent,and their mechanism may be related to the in-hibition of the TLR4/Myd88/NF-κB signaling pathway.
张红霞;王莉平;曹亚茹;冯梅梅
石家庄市第四医院,石家庄 050010邯郸市中心医院,邯郸 056001石家庄市第四医院,石家庄 050010石家庄市第四医院,石家庄 050010
多囊卵巢综合征银杏二萜内酯卵巢功能炎症Toll样受体4/髓样分化因子88/核因子-κB信号通路
Polycystic ovary syndromeGinkgo diterpene lactoneOvarian functionInflammationToll-like receptor 4/myeloid differentiation factor 88/nuclear factor kappa B signaling pathway
《中药药理与临床》 2026 (4)
51-58,8
邯郸市科学技术研究与发展计划项目(编号:23422083221)石家庄市科学技术研究与发展指导计划课题(编号:141463073).
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