G蛋白偶联受体120调控NLRP3炎症小体参与多囊卵巢综合征炎症反应的分子机制探究  

Molecular Mechanism of G Protein-Coupled Receptor 120 Regulate NLRP3 Inflammasome-Mediated Inflammatory Response in Polycystic Ovary Syndrome

作　　者：阳丽[1] 吴湘[1] 李昂[1] 吴菲[1] 何薇薇[1] 张珂[1] YANG Li;WU Xiang;LI Ang(Department of Gynaecology,Hunan Maternal and Child Health Hospital,Changsha Hunan 410008,China)

机构地区：[1]湖南省妇幼保健院妇一科,湖南长沙410008

出　　处：《实用妇产科杂志》2025年第3期216-222,共7页Journal of Practical Obstetrics and Gynecology

基　　金：湖南省卫生健康委员会科研项目(编号:20210124)。

摘　　要：目的:探讨G蛋白偶联受体120(GPR120)调控核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体介导多囊卵巢综合征(PCOS)炎症反应的机制。方法:45只野生型雌性C57BL/6J小鼠随机分为:对照组、模型组、TUG-891组,每组15只。模型组和TUG-891组皮下植入35 d睾酮(T)连续释放药丸诱导PCOS模型,对照组接受安慰剂药丸。TUG-891组小鼠在植入T 21 d后每天腹腔注射TUG-891治疗,模型组与对照组腹腔注射等量0.9%氯化钠液。实验结束时(第35天)处死小鼠,收集血液样本和卵巢组织。通过免疫荧光染色检测卵巢组织中GPR120、NLRP3蛋白表达,天狼星红染色检测卵巢纤维化,免疫印迹分析检测组织中纤维化因子α-平滑肌肌动蛋白(α-SMA)、基质金属蛋白酶2(MMP2)、转化生长因子-β1(TGF-β1)和炎症因子NLRP3、Toll样受体4(TLR4)和肿瘤坏死因子-α(TNF-α)表达。从雌性小鼠中分离卵巢颗粒细胞(GCs),原代GCs接种在含10%胎牛血清和1%青霉素-链霉素溶液的DMEM-F12中培养。将细胞分为对照组、TUG-891组、T组和T+TUG-891组。分别通过MTT法和流式细胞术分析GCs活力和凋亡水平。结果:与模型组比较,TUG-891组小鼠的体质量、卵巢重量、卵巢系数和排卵前卵泡数显著增加(P<0.05),以及性激素T、雌二醇(E 2)、促黄体生成素(LH)、卵泡刺激素(FSH)水平和闭锁卵泡数量显著降低(P<0.05)。TUG-891组卵巢组织中天狼星红染色和α-SMA、MMP2、TGF-β1的相对表达较模型组显著降低(P<0.05)。与模型组比较,TUG-891组卵巢组织中GRP120表达显著增加(P<0.05)、NLRP3表达显著减弱(P<0.05)。体外实验中,与T组比较,T+TUG-891组GCs增殖明显增加(P<0.05),以及GCs凋亡和GCs中NLRP3、TLR4和TNF-α蛋白表达明显减少(P<0.05)。结论:GPR120激动剂TUG-891可能通过抑制NLRP3炎性体的产生,减轻高雄激素血症诱导的PCOS小鼠慢性轻度炎症,进而保护小鼠的卵巢和卵泡的生长发育能力。Objective:To explore the mechanism of G protein-coupled receptor 120(GPR120)regulating nod-like receptor family pyrin domain containing 3(NLRP3)inflammasome-mediated inflammatory response in polycystic ovary syndrome(PCOS).Methods:Forty-five wild-type female C57BL/6J mice were randomly divided into three groups:control group,model group,and TUG-891 group,with 15 mice in each group.The model and TUG-891 groups were subcutaneously implanted with 35-day testosterone(T)continuous-release pellets to induce the PCOS model,while control mice received placebo pills.Mice in the TUG-891 group were treated with daily intraperitoneal injections of TUG-891 starting from day 21 after T implantation.The model group and the control group were injected with the same amount of normal saline intraperitoneally.At the end of the experiment(day 35),the mice were euthanized,and blood and ovarian tissue samples were collected.Immunofluorescence staining was used to detect the expression of GPR120 and NLRP3 proteins in ovarian tissues.Sirius Red staining was performed to detect ovarian fibrosis.Western blot analysis was used to measure the expression of fibrosis factors:α-smooth muscle actin(α-SMA),matrix metalloproteinase 2(MMP2),transforming growth factor-β1(TGF-β1),and inflammatory factors NLRP3,Toll-like receptor 4(TLR4),and tumor necrosis factor-α(TNF-α)in tissues.Ovarian granulosa cells(GCs)were isolated from female mice and cultured in DMEM-F12 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin.The cells were divided into the control group,TUG-891 group,T group,and T+TUG-891 group.The vitality and apoptosis of GCs were analyzed using the MTT assay and flow cytometry,respectively.Results:Compared with the model group,the TUG-891 group showed significant increases in body growth,ovarian weight,ovarian index,and the number of pre-ovulatory follicles(P<0.05),as well as significant reductions in sex hormones T,estradiol(E 2),luteinizing hormone(LH),and follicle-stimulating hormone(FSH)levels and atretic folli

关 键 词：G蛋白偶联受体120 核苷酸结合寡聚化结构域样受体蛋白3 多囊卵巢综合征 炎症 

分 类 号：R711.75[医药卫生—妇产科学]