二甲双胍对多囊卵巢综合征模型大鼠炎症的影响及其机制研究

doi:10.12280/gjfckx.20250895

摘要/Abstract

摘要：

目的：探讨二甲双胍对多囊卵巢综合征（polycystic ovary syndrome，PCOS）模型大鼠炎症反应的作用及对丝裂原激活的蛋白激酶（mitogen-activated protein kinase，MAPK）/核因子κB（nuclear factor-κB，NF-κB）通路的调控作用。方法：50只雌性SD大鼠中，40只用来曲唑诱导PCOS模型后随机分为模型组和二甲双胍低、中、高剂量组（均n=10）；余10只为对照组。二甲双胍低、中、高剂量组分别给予50、100、200 mg/kg二甲双胍灌胃（体积均为5 mL/kg），对照组和模型组给予5 mL/kg生理盐水灌胃，各组均1次/d，连续4周。观察动情周期变化；采用酶联免疫吸附分析（enzyme-linked immunosorbent assay，ELISA）检测血清性激素[卵泡刺激素（follicle-stimulating hormone，FSH）、黄体生成素（luteinizing hormone，LH）、雌二醇（estradiol，E₂）、睾酮（testosterone，T）]及炎症因子[肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、白细胞介素-6（interleukin-6，IL-6）、IL-18]水平；HE染色观察卵巢形态；TUNEL染色观察卵巢细胞凋亡；蛋白质印迹（Western blotting）法检测卵巢组织中细胞外信号调节激酶（extracellular signal-regulated kinase，ERK）、磷酸化ERK（p-ERK）、c-Jun氨基端蛋白激酶（c-Jun N-terminal protein kinase，JNK）、p-JNK、p38 MAPK、p-p38 MAPK、NF-κB p65及p-NF-κB p65蛋白表达。结果：模型组动情周期紊乱，各给药组动情周期均有改善，二甲双胍高剂量组改善最显著。与对照组相比，模型组血清FSH、E₂水平降低，LH、T、炎症因子、细胞凋亡率及p-ERK、p-JNK、p-p38 MAPK、p-NF-κB p65蛋白表达水平升高（均P<0.05）；与模型组比较，各二甲双胍给药组FSH、E₂水平升高，LH、T、炎症因子、细胞凋亡率及p-ERK、p-JNK、p-p38 MAPK、p-NF-κB p65蛋白表达水平降低，且以二甲双胍高剂量组改善最显著（均P<0.05）。各给药组卵巢形态学异常改善，二甲双胍高剂量组改善最显著。结论：二甲双胍可改善PCOS大鼠动情周期，调节性激素，减少细胞凋亡，可能通过抑制MAPK/NF-κB通路缓解炎症反应发挥作用。

关键词: 多囊卵巢综合征, 大鼠, 炎症, 丝裂原激活蛋白激酶类, 核因子κB

Abstract:

Objective: To investigate the effect of metformin on the inflammatory response in polycystic ovary syndrome (PCOS) model rats and its regulatory role in the mitogen-activated protein kinase (MAPK)/nuclear factor-κB (NF-κB) pathway. Methods: Among 50 female SD rats, 40 rats were used to establish a PCOS model via letrozole induction and then randomly divided into the model group and low-, medium-, and high-dose metformin groups (n=10 each). The remaining 10 rats served as the control group. The low-, medium- and high-dose metformin groups were administered metformin via oral gavage at doses of 50 mg/kg, 100 mg/kg, and 200 mg/kg, respectively (all at a volume of 5 mL/kg). The control and model groups received 5 mL/kg normal saline via oral gavage. All treatments were administered once daily for four consecutive weeks. The estrous cycle changes were observed. Serum sex hormone levels [follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E₂), testosterone (T)] and inflammatory factor levels [tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-18] were measured by enzyme-linked immunosorbent assay (ELISA). Ovarian morphology was examined via hematoxylin and eosin (HE) staining, and ovarian cell apoptosis was assessed via TUNEL staining. The expression levels of extracellular signal-regulated kinase (ERK), phosphorylated ERK (p-ERK), c-Jun N-terminal protein kinase (JNK), p-JNK, p38 MAPK, p-p38 MAPK, NF-κB p65, and p-NF-κB p65 proteins in ovarian tissues were detected by Western blotting. Results: The estrous cycle was disordered in the model group, while all metformin-treated groups showed improvements, with the most significant improvement observed in the high-dose metformin group. Compared with the control group, the model group exhibited decreased serum levels of FSH and E₂, and increased levels of LH, T, inflammatory factors, apoptosis rate, and the expression of p-ERK, p-JNK, p-p38 MAPK and p-NF-κB p65 proteins (all P<0.05). Compared with the model group, all metformin-treated groups demonstrated increased levels of FSH and E₂, and decreased levels of LH, T, inflammatory factors, apoptosis rate, and the expression of p-ERK, p-JNK, p-p38 MAPK and p-NF-κB p65 proteins, with the most significant improvements observed in the high-dose metformin group (all P<0.05). Ovarian morphology abnormalities improved in all metformin-treated groups, most notably in the high-dose metformin group. Conclusions: Metformin can improve the estrous cycle, regulate sex hormones, and reduce cell apoptosis in PCOS rats. Its mechanism of action may involve alleviating the inflammatory response by inhibiting the MAPK/NF-κB pathway.

Key words: Polycystic ovary syndrome, Rats, Inflammation, Mitogen-activated protein kinases, Nuclear factor-κB

赵兴敏, 刘艳萍, 高争, 张国庆. 二甲双胍对多囊卵巢综合征模型大鼠炎症的影响及其机制研究[J]. 国际妇产科学杂志, 2026, 53(2): 143-149.

ZHAO Xing-min, LIU Yan-ping, GAO Zheng, ZHANG Guo-qing. The Effect and Mechanism of Metformin on Inflammation in Polycystic Ovary Syndrome Model Rats[J]. Journal of International Obstetrics and Gynecology, 2026, 53(2): 143-149.

图/表 7

参考文献 21

[1]	Siddiqui S, Mateen S, Ahmad R, et al. A brief insight into the etiology, genetics, and immunology of polycystic ovarian syndrome (PCOS)[J]. J Assist Reprod Genet, 2022, 39(11):2439-2473. doi: 10.1007/s10815-022-02625-7.
[2]	Armanini D, Boscaro M, Bordin L, et al. Controversies in the Pathogenesis, Diagnosis and Treatment of PCOS: Focus on Insulin Resistance, Inflammation, and Hyperandrogenism[J]. Int J Mol Sci, 2022, 23(8):4110. doi: 10.3390/ijms23084110.
[3]	Liu K, He X, Huang J, et al. Short-chain fatty acid-butyric acid ameliorates granulosa cells inflammation through regulating METTL3-mediated N6-methyladenosine modification of FOSL2 in polycystic ovarian syndrome[J]. Clin Epigenetics, 2023, 15(1):86. doi: 10.1186/s13148-023-01487-9. pmid: 37179374
[4]	Guo Y, Zhang H, Lv Z, et al. Up-regulated CD38 by daphnetin alleviates lipopolysaccharide-induced lung injury via inhibiting MAPK/NF-κB/NLRP3 pathway[J]. Cell Commun Signal, 2023, 21(1):66. doi: 10.1186/s12964-023-01041-3. pmid: 36998049
[5]	Zhao X, Li M, Lu Y, et al. Sirt1 inhibits macrophage polarization and inflammation in gouty arthritis by inhibiting the MAPK/NF-κB/AP-1 pathway and activating the Nrf2/HO-1 pathway[J]. Inflamm Res, 2024, 73(7):1173-1184. doi: 10.1007/s00011-024-01890-9. pmid: 38739197
[6]	Zhao H, Xing C, Zhang J, et al. Comparative efficacy of oral insulin sensitizers metformin, thiazolidinediones, inositol, and berberine in improving endocrine and metabolic profiles in women with PCOS: a network meta-analysis[J]. Reprod Health, 2021, 18(1):171. doi: 10.1186/s12978-021-01207-7. pmid: 34407851
[7]	Kukaev E, Kirillova E, Tokareva A, et al. Impact of Gut Microbiota and SCFAs in the Pathogenesis of PCOS and the Effect of Metformin Therapy[J]. Int J Mol Sci, 2024, 25(19):10636. doi: 10.3390/ijms251910636.
[8]	Balasubramanian A, Pachiappan S, Mohan S, et al. Therapeutic exploration of polyherbal formulation against letrozole induced PCOS rats: A mechanistic approach[J]. Heliyon, 2023, 9(5):e15488. doi: 10.1016/j.heliyon.2023.e15488.
[9]	Dabravolski SA, Nikiforov NG, Eid AH, et al. Mitochondrial Dysfunction and Chronic Inflammation in Polycystic Ovary Syndrome[J]. Int J Mol Sci, 2021, 22(8):3923. doi: 10.3390/ijms22083923.
[10]	袁山政, 张燕雨, 翟亚悦, 等. 循环炎症因子与PCOS风险之间的关联评估:一项双样本孟德尔随机化研究[J]. 生殖医学杂志, 2024, 33(9):1214-1220. doi: 10.3969/j.issn.1004-3845.2024.09.013.
[11]	刘聪, 杜宇坤, 折慧芝, 等. 二甲双胍联合枸橼酸氯米芬治疗对PCOS合并不孕症患者胰岛素、HOMA-IR水平及卵巢排卵功能的影响[J]. 生殖医学杂志, 2025, 34(4):430-435. doi: 10.3969/j.issn.1004-3845.2025.04.002.
[12]	Kumari R, Singh A, Banerjee BD, et al. Impact of Metformin Therapy on Chronic Inflammatory Markers Serum Fractalkine and CRP Levels in Polycystic Ovary Syndrome: A Pilot Study on Chronic Inflammatory Markers and Clinical Outcomes[J]. J Obstet Gynaecol India, 2025, 75(2):115-121. doi: 10.1007/s13224-025-02100-0.
[13]	Yousuf SD, Ganie MA, Urwat U, et al. Oral contraceptive pill (OCP) treatment alters the gene expression of intercellular adhesion molecule-1 (ICAM-1), tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1) and plasminogen activator inhibitor-1 (PAI-1) in polycystic ovary syndrome (PCOS) women compared to drug-naive PCOS women[J]. BMC Womens Health, 2023, 23(1):68. doi: 10.1186/s12905-023-02187-5. pmid: 36793022
[14]	Bansal B, Thazhuthadath Kishore A, Kathiresan S, et al. A Systematic Review of Inflammatory Markers in Polycystic Ovary Syndrome (PCOS) and Meta-Analysis of Interleukin-6 (IL-6) in Case-Control Studies[J]. Cureus, 2025, 17(4):e82350. doi: 10.7759/cureus.82350.
[15]	朱松楠, 张红媛, 李艳. 血清NLRP3、IL-18水平与多囊卵巢综合征患者不孕的关系[J]. 中国现代医学杂志, 2024, 34(13):1-6. doi: 10.3969/j.issn.1005-8982.2024.13.001.
[16]	符山花, 包利利, 赵达, 等. 抑制NLRP3炎症小体激活可调节自噬改善多囊卵巢综合征颗粒细胞凋亡[J]. 中国免疫学杂志, 2024, 40(8):1646-1652. doi: 10.3969/j.issn.1000-484X.2024.08.013.
[17]	Liu Z, Wang RH, Wang KH. Formononetin ameliorates polycystic ovary syndrome through suppressing NLRP3 inflammasome[J]. Mol Med, 2025, 31(1):27. doi: 10.1186/s10020-025-01092-x. pmid: 39871124
[18]	李春颖, 尹燕燕, 季梦垚, 等. 基于MAPK/NF-κB信号通路探讨调气止咳方对咳嗽变异性哮喘大鼠气道炎症的影响及机制[J]. 中药新药与临床药理, 2024, 35(7):1008-1015. doi: 10.19378/j.issn.1003-9783.2024.07.009.
[19]	柯玲玲, 王敏, 高洁, 等. STAT1对肺结核大鼠巨噬细胞凋亡及MAPK/NF-κB信号通路的机制研究[J]. 解剖学研究, 2024, 46(2):115-122. doi: 10.20021/j.cnki.1671-0770.2024.02.04.
[20]	曹静, 刘海波, 安琪, 等. 二甲双胍缓解小鼠放射性皮炎引起的病理性疼痛:基于抑制p38 MAPK/NF-κB信号通路[J]. 南方医科大学学报, 2023, 43(10):1815-1820. doi: 10.12122/j.issn.1673-4254.2023.10.22.
[21]	黄辉, 于大海, 黄炫赓, 等. 二甲双胍对糖基化终末产物诱导牙周膜成纤维细胞凋亡及p38 MAPK/NF-κB通路的影响[J]. 山东医药, 2021, 61(14):19-23. doi: 10.3969/j.issn.1002-266X.2021.14.005.

组别	n	FSH（ng/mL）	LH（mg/mL）	E₂（pg/mL）	T（ng/mL）
对照组	10	6.34±0.54	8.42±0.92	182.59±15.58	1.96±0.23
模型组	10	2.96±0.39^a	21.46±1.58^a	124.79±10.73^a	3.49±0.31^a
二甲双胍低剂量组	10	3.75±0.42^ab	17.48±1.33^ab	139.68±11.85^ab	3.01±0.29^ab
二甲双胍中剂量组	10	4.46±0.48^abc	14.26±1.29^abc	152.36±12.96^abc	2.71±0.26^abc
二甲双胍高剂量组	10	5.57±0.53^abcd	11.38±1.11^abcd	166.96±13.38^abcd	2.27±0.25^abcd
F		81.570	162.299	30.220	50.387
P		<0.001	<0.001	<0.001	<0.001

组别	n	FSH（ng/mL）	LH（mg/mL）	E₂（pg/mL）	T（ng/mL）
对照组	10	6.34±0.54	8.42±0.92	182.59±15.58	1.96±0.23
模型组	10	2.96±0.39^a	21.46±1.58^a	124.79±10.73^a	3.49±0.31^a
二甲双胍低剂量组	10	3.75±0.42^ab	17.48±1.33^ab	139.68±11.85^ab	3.01±0.29^ab
二甲双胍中剂量组	10	4.46±0.48^abc	14.26±1.29^abc	152.36±12.96^abc	2.71±0.26^abc
二甲双胍高剂量组	10	5.57±0.53^abcd	11.38±1.11^abcd	166.96±13.38^abcd	2.27±0.25^abcd
F		81.570	162.299	30.220	50.387
P		<0.001	<0.001	<0.001	<0.001

组别	n	TNF-α	IL-6	IL-18
对照组	10	42.89±4.24	53.52±5.42	19.89±1.24
模型组	10	83.76±7.61^a	117.15±9.69^a	43.75±3.57^a
二甲双胍低剂量组	10	72.23±6.31^ab	96.43±9.11^ab	36.23±3.31^ab
二甲双胍中剂量组	10	61.68±5.11^abc	83.14±8.58^abc	29.68±2.07^abc
二甲双胍高剂量组	10	52.21±4.85^abcd	67.46±6.47^abcd	23.21±1.85^abcd
F		78.512	95.394	142.592
P		<0.001	<0.001	<0.001

组别	n	TNF-α	IL-6	IL-18
对照组	10	42.89±4.24	53.52±5.42	19.89±1.24
模型组	10	83.76±7.61^a	117.15±9.69^a	43.75±3.57^a
二甲双胍低剂量组	10	72.23±6.31^ab	96.43±9.11^ab	36.23±3.31^ab
二甲双胍中剂量组	10	61.68±5.11^abc	83.14±8.58^abc	29.68±2.07^abc
二甲双胍高剂量组	10	52.21±4.85^abcd	67.46±6.47^abcd	23.21±1.85^abcd
F		78.512	95.394	142.592
P		<0.001	<0.001	<0.001

组别	n	p-ERK	p-JNK	p-p38 MAPK	p-NF-κB p65
对照组	10	0.21±0.03	0.19±0.02	0.26±0.04	0.28±0.03
模型组	10	1.07±0.09^a	0.94±0.08^a	0.84±0.07^a	0.91±0.09^a
二甲双胍低剂量组	10	0.82±0.06^ab	0.76±0.07^ab	0.71±0.06^ab	0.77±0.07^ab
二甲双胍中剂量组	10	0.69±0.05^abc	0.57±0.05^abc	0.57±0.05^abc	0.58±0.06^abc
二甲双胍高剂量组	10	0.43±0.04^abcd	0.38±0.04^abcd	0.42±0.04^abcd	0.46±0.05^abcd
F		324.005	253.897	168.355	149.563
P		<0.001	<0.001	<0.001	<0.001