蛇床子素通过Nrf2/HO-1/NLRP3轴对大鼠卵巢储备功能减退的改善作用研究

Study on the Improvement Effects of Osthole on Diminished Ovarian Reserve Rats by Nrf2/HO-1/NLRP3 Axis

目的:探讨蛇床子素(OST)对卵巢储备功能减退(DOR)大鼠的改善作用及其可能机制。方法:取80只大鼠,64只以50 mg·(kg·d)^(-1)雷公藤多苷混悬液灌胃建立DOR大鼠模型,随机分为模型组、OST低剂量组、OST中剂量组、OST高剂量组,每组16只大鼠,其余16只为空白组。OST低、中、高剂量组按照2 mL·d^(-1)分别腹腔注射20、40和80μg·mL^(-1) OST溶液,空白组、模型组注射等体积生理盐水,1次·d^(-1),给药21 d。比较各组大鼠卵巢组织学形态变化、脏器指数和血清中抗苗勒管激素(AMH)、雌二醇激素(E2)、促黄体生成激素(LH)、卵泡刺激素(FSH)、白细胞介素4(IL4)、γ干扰素(IFN-γ)、肿瘤坏死因子-α(TNF-α)、白细胞介素10(IL-10)水平,以及卵巢组织中超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性、丙二醛(MDA)水平和转录因子核因子E2相关因子2(Nrf2)、血红素氧合酶1(HO-1)、核苷酸结合寡聚化结构域受体3(NLRP3)蛋白表达水平。结果:HE染色结果显示,OST各剂量组卵巢组织卵泡和黄体数较模型组增加;与模型组比,OST各剂量组AMH、E2、IL-4、IL-10水平,卵巢指数,子宫指数,SOD、CAT活性和Nrf2、HO-1蛋白表达水平均升高,LH、FSH、IFN-γ、TNF-α、MDA水平和NLRP3蛋白表达水平均降低(P<0.05)。AMH、E2、IL-4、IL-10水平,卵巢指数,子宫指数,SOD、CAT活性和Nrf2、HO-1蛋白表达水平与OST呈剂量依赖性升高,LH、FSH、IFN-γ、TNF-α、MDA水平和NLRP3蛋白表达水平与OST呈剂量依赖性降低(P<0.05)。结论:OST可有效改善DOR大鼠内分泌代谢,减轻卵巢炎症反应和氧化应激损伤,恢复卵巢储备功能,可能通过调控Nrf2/HO-1/NLRP3信号通路发挥作用。

Objective: To investigate the improvement ef fect of osthole(OST) on diminished ovarian reserve(DOR) rats and its possible mechanism. Methods: Eighty rats were taken as experimental subjects, 64 of which were made DOR rat model by intragastric administration of 50 mg·(kg·d)^(-1) tripterygium glycosides suspension, randomly divided into model group, low dose of OST group, medium dose of OST group and high dose of OST group, there were 16 rats in each group, and the remaining 16 rats were blank group. The low, medium and high dose of OST groups were intraperitoneally injected with 20, 40 and 80 μg·mL^(-1) OST solution at 2 mL·d^(-1), the blank group and model group were injected with equal volume of saline, once per day, for 21 days. The morphological changes of the ovaries, organ index, the levels of anti-Müllerian hormone(AMH), estradiol(E2), luteinizinghormone(LH), follicle stimulating hormone(FSH), interleukin-4(IL-4),interferon-γ(IFN-γ), tumor necrosis factor-α(TNF-α), interleukin-10(IL-10) in serum, the activity of superoxide dismutase(SOD) and catalase(CAT), malondialdehyde(MDA) level and the expression levels of nuclear factor ervthroicl2-related factor 2(Nrf2), hemeoxygenasel-1(HO-1), nucleotide-binding oligomerization domainlike receptor protein 3(NLRP3) protein in ovarian tissue were compared. Results: The results of HE staining showed that the number of follicles and corpus luteum in the ovarian tissue of each dose group of OST increased compared with the model group. Compared with the model group, the levels of AMH, E2, IL-4, IL-10, ovarian index, uterine index, the activity of SOD and CAT, the expression levels of Nrf2, HO-1 protein increased, while the levels of LH, FSH, IFN-γ, TNF-α, MDA and the expression level of NLRP3 protein decreased in each dose group of OST(P<0.05). AMH, E2, IL-4, IL-10 levels, ovarian index, uterine index, the activity of SOD and CAT, the expression levels of Nrf2, HO-1 protein increased in a dose-dependent manner with OST, the levels of LH, FSH, IFN-γ, TNF-α, MDA and the expression level of NLRP3 protein decreased in a dose-dependent manner with OST(P<0.05). Conclusion: OST could effectively improve endocrine metabolism in DOR rats, reduce ovarian infl ammation and oxidative stress damage, and restore ovarian reserve function, which may play a role by regulating the Nrf2/HO-1/NLRP3 signaling pathway.