目的 探究大豆异黄酮（soy isoflavone, SIF）对高脂饲料诱导的肥胖小鼠慢性炎症的缓解作用,并阐明其调控脂肪组织巨噬细胞极化的信号通路机制。方法 将30只C57BL/6小鼠随机分为对照组、模型组、10 mg/kg SIF组、20 mg/kg SIF组和罗格列酮组（阳性药）。除对照组外均给予高脂饲料喂养12 w;检测小鼠体质量、附睾脂肪组织重量、血清总胆固醇（total cholesterol, TC）、甘油三酯（triglyceride, TG）、低密度脂蛋白胆固醇（low-density lipoprotein cholesterol, LDL-C）水平及附睾脂肪组织炎性因子水平,对模型组与20 mg/kg SIF组脂肪组织进行转录组测序及GO/KEGG富集分析。采用免疫组化染色检测脂肪组织CD86阳性率,Western blot检测脂肪组织中iNOS、CD86及过氧化物酶体增殖物激活受体γ（peroxisome proliferator-activated receptor γ, PPARγ）蛋白表达。体外培养RAW264.7巨噬细胞,分为对照组、模型组、10 μmol/L SIF、30μmol/L SIF 和30 μmol/L SIF+PPARγ抑制剂GW9662（10 μmol/L）组;除对照组外,各组经LPS诱导M1极化;采用流式细胞术检测M1型极化比例,Western blot检测iNOS、CD86及PPARγ表达,ELISA检测RAW264.7巨噬细胞炎性因子水平,通过分子对接和表面等离子体共振（surface plasmon resonance,SPR）试验验证SIF与PPARγ的相互作用。结果 与模型组相比,SIF干预可降低肥胖小鼠体质量、附睾脂肪组织质量及血清TC、TG、LDL-C水平,下调脂肪组织炎症因子水平;转录组测序显示,SIF干预后附睾脂肪组织中“炎症反应”“巨噬细胞M1极化”相关GO条目及PPARγ信号通路显著富集;免疫组化结果显示SIF可降低附睾脂肪组织CD86阳性率,下调附睾脂肪组织iNOS、CD86表达,上调PPARγ表达。体外实验表明,SIF可降低LPS诱导的RAW264.7细胞M1型极化比例,下调iNOS、CD86蛋白表达及TNF-α、IL-1β、IL-6水平,而 GW9662可逆转上述效应。分子对接显示SIF与PPARγ具有良好结合亲和力,SPR实验进一步证实两者存在直接相互作用（Kd值为1.2 μmol/L）。结论 SIF可直接靶向结合并激活PPARγ,进而抑制脂肪组织巨噬细胞M1型极化,缓解肥胖相关慢性炎症。
Abstract
Objective To investigate the alleviating effects of soy isoflavones (SIF) on chronic inflammation in obese mice induced by a high-fat diet and elucidate the molecular mechanisms Methods Thirty C57BL/6 mice were randomly divided into control group, model group, 10 mg/kg SIF group, 20 mg/kg SIF group, and rosiglitazone group (positive control). All groups except the control group were fed a high-fat diet for 12 weeks. Body weight, epididymal adipose tissue weight, serum levels of total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C), as well as inflammatory factor in adipose tissue, were measured. Transcriptome sequencing and GO/KEGG enrichment analysis were performed on epididymal adipose tissues from the model group and the 20 mg/kg SIF group. Immunohistochemical staining was used to detect the positive rate of CD86 in epididymal adipose tissue, and Western blot was employed to assess the protein expression of iNOS, CD86, and PPARγ in epididymal adipose tissue. RAW264.7 macrophages were cultured in vitro and divided into control group, model group, 10 μmol/L SIF, 30 μmol/L SIF, and 30 μmol/L SIF + PPARγ inhibitor GW9662 (10 μmol/L)groups. Except for the control group, M1 polarization was induced by LPS in all groups. The proportion of M1-polarized cells was detected by flow cytometry, and the expression levels of iNOS, CD86, and PPARγ were assessed by Western blot. Inflammatory factors levels in RAW264.7 macrophages were measured by ELISA. Molecular docking and surface plasmon resonance (SPR) assays were performed to validate the interaction between SIF and PPARγ. Results Compared with the model group, SIF intervention reduced body weight, epididymal adipose tissue weight, serum levels of TC, TG, and LDL-C, and inflammatory factors levels in epididymal adipose tissue. Transcriptome sequencing revealed that after SIF intervention, GO terms related to inflammatory response and M1 macrophage polarization,as well as the PPARγ signaling pathway, were significantly enriched in epididymal adipose tissue. Immunohistochemical results showed that SIF reduced the positive rate of CD86 in epididymal adipose tissue. SIF downregulated the expression of iNOS and CD86, while upregulating PPARγ expression in epididymal adipose tissue. In vitro experiments demonstrated that SIF reduced the proportion of LPS-induced M1-polarized RAW264.7 cells, downregulated the protein expression of iNOS and CD86, and decreased the levels of TNF-α, IL-1β, and IL-6. These effects were reversed by GW9662. Molecular docking indicated a favorable binding affinity between SIF and PPARγ, and SPR assays further confirmed their direct interaction (Kd value of 1.2 μmol/L). Conclusion SIF can inhibit M1 polarization of tissue macrophages in adipose tissue, thereby alleviating obesity-related chronic inflammation. The mechanism is associated with direct binding to and activation of PPARγ.
关键词
肥胖 /
大豆异黄酮 /
慢性炎症 /
巨噬细胞 /
PPARγ /
小鼠
Key words
obesity /
soy isoflavones /
chronic inflammation /
macrophages /
PPARγ /
mouse
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] Kopczyńska J, Kowalczyk M.The potential of short-chain fatty acid epigenetic regulation in chronic low-grade inflammation and obesity[J]. Front Immunol, 2024, 15:1380476.
[2] Cruz KJC, de Oliveira ARS, Morais JBS, et al. Role of microRNAs on adipogenesis, chronic low-grade inflammation, and insulin resistance in obesity[J]. Nutrition, 2017, 35:28–35.
[3] 吴欧, 芈肖肖, 张虎, 等. TLR5在肥胖相关高血压发病机制中的作用:现有证据和未来挑战[J]. 中国免疫学杂志, 2025, 41:484-489.
[4] Allocca S, Monda A, Messina A, et al.Endocrine and metabolic mechanisms linking obesity to type 2 diabetes: implications for targeted therapy[J]. Healthcare (Basel), 2025, 13:1437.
[5] Hu F, Tong J, Deng B, et al.MiR-495 regulates macrophage M1/M2 polarization and insulin resistance in high-fat diet-fed mice via targeting FTO[J]. Pflugers Arch, 2019, 471:1529–1537.
[6] Dousdampanis P, Aggeletopoulou I, Mouzaki A.The role of M1/M2 macrophage polarization in the pathogenesis of obesity-related kidney disease and related pathologies[J]. Front Immunol, 2024, 15:1534823.
[7] Chylikova J, Dvorackova J, Tauber Z, et al.M1/M2 macrophage polarization in human obese adipose tissue[J]. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub, 2018, 162:79–82.
[8] Pawlak M, Lefebvre P, Staels B.Molecular mechanism of PPARα action and its impact on lipid metabolism, inflammation and fibrosis in non-alcoholic fatty liver disease[J]. J Hepatol, 2015, 62:720–733.
[9] Yang XF, Shang DJ.The role of peroxisome proliferator-activated receptor γ in lipid metabolism and inflammation in atherosclerosis[J]. Cell Biol Int, 2023, 47:1469–1487.
[10] Lin G, Lin L, Chen X, et al.PPAR-γ/NF-kB/AQP3 axis in M2 macrophage orchestrates lung adenocarcinoma progression by upregulating IL-6[J]. Cell Death Dis, 2024, 15:532.
[11] Balfour JA, Plosker GL.Rosiglitazone[J]. Drugs, 1999, 5:921–930.
[12] Zhou L, Liu G, Jia Z, et al.Increased susceptibility of db/db mice to rosiglitazone-induced plasma volume expansion: role of dysregulation of renal water transporters[J]. Am J Physiol Renal Physiol, 2013, 305:F1491–F1497.
[13] Cheng D, Gao H, Li W.Long-term risk of rosiglitazone on cardiovascular events - a systematic review and meta-analysis[J]. Endokrynol Pol, 2018, 6:381-394.
[14] Tolman KG.The safety of thiazolidinediones[J]. Expert Opin Drug Saf, 2011, 1:419-428.
[15] Harsløf T, Wamberg L, Møller L, et al.Rosiglitazone decreases bone mass and bone marrow fat[J]. J Clin Endocrinol Metab, 2011, 9:1541–1548.
[16] Akhlaghi M, Zare M, Nouripour F.Effect of soy and soy isoflavones on obesity-related anthropometric measures: a systematic review and meta-analysis of randomized controlled clinical trials[J]. Adv Nutr, 2017, 8:705–717.
[17] Zhu J, Zhao Q, Qiu Y, et al.Soy isoflavones intake and obesity in Chinese adults: a cross-sectional study in Shanghai, China[J]. Nutrients, 2021, 13:2715.
[18] Barańska A, Błaszczuk A, Polz-Dacewicz M, et al.Effects of soy isoflavones on glycemic control and lipid profile in patients with type 2 diabetes: a systematic review and meta-analysis of randomized controlled trials[J]. Nutrients, 2021, 13:1886.
[19] Zhang J, Zhu Y, Pan L, et al.Soy isoflavone improved female sexual dysfunction of mice via endothelial nitric oxide synthase pathway[J]. Sex Med, 2019, 7:345-351.
[20] Nagarajan S, Stewart BW, Badger TM.Soy isoflavones attenuate human monocyte adhesion to endothelial cell-specific CD54 by inhibiting monocyte CD11a[J]. J Nutr, 2006, 136:2384–2390.
[21] Engin A.Reappraisal of adipose tissue inflammation in obesity[J]. Adv Exp Med Biol, 2024, 1460:297–327.
[22] Ren Y, Zhao H, Yin C, et al.Adipokines, hepatokines and myokines: focus on their role and molecular mechanisms in adipose tissue inflammation[J]. Front Endocrinol (Lausanne), 2022, 13:873699.
[23] Shapouri-Moghaddam A, Mohammadian S, Vazini H, et al.Macrophage plasticity, polarization, and function in health and disease[J]. J Cell Physiol, 2018, 233:6425–6440.
[24] Huang C, Pang D, Luo Q, et al.Soy isoflavones regulate lipid metabolism through an AKT/mTORC1 pathway in diet-induced obesity (DIO) male rats[J]. Molecules, 2016, 21:586.
[25] Llaneza P, González C, Fernández-Iñarrea J, et al.Soy isoflavones improve insulin sensitivity without changing serum leptin among postmenopausal women[J]. Climacteric, 2012, 15:611–620.
[26] 邵国权, 明哲, 王文晟, 等. 大豆异黄酮改善代谢紊乱的研究进展[J]. 食品工业科技, 2026, 47:1–11.
[27] 苑慧敏, 李小琼, 朱立颖, 等. 大豆异黄酮对2型糖尿病小鼠脂代谢及肠道菌群结构的影响[J]. 营养学报, 2021, 43:556–565.
[28] 李立科, 罗启慧, 唐秀莹, 等. 大豆异黄酮对肥胖大鼠肠道瘦素介导Janus激酶/信号转导及转录激活因子信号转导通路的影响[J]. 动物营养学报, 2016, 28:2448–2457.
[29] Tan J, Huang C, Luo Q, et al.Soy isoflavones ameliorate fatty acid metabolism of visceral adipose tissue by increasing the AMPK activity in male rats with diet-induced obesity (DIO)[J]. Molecules, 2019, 24:2809.
[30] Kim SE, Kawaguchi K, Hayashi H, et al.Remission effects of dietary soybean isoflavones on DSS-induced murine colitis and an LPS-activated macrophage cell line[J]. Nutrients, 2019, 11:1746.
[31] Abernathy LM, Fountain MD, Rothstein SE, et al.Soy isoflavones promote radioprotection of normal lung tissue by inhibition of radiation-induced activation of macrophages and neutrophils[J]. J Thorac Oncol, 2015, 10:1703–1712.
[32] de Ganuza CR, López S, Mendoza G. Exploring the anti-inflammatory effects of genistein in an in vitro lipopolysaccharide-induced macrophage model[J]. Sci Rep, 2026, 16:11592.
[33] Xu Y, Zhang D, Yang H, et al.Hepatoprotective effect of genistein against dimethylnitrosamine-induced liver fibrosis in rats by regulating macrophage functional properties and inhibiting the JAK2/STAT3/SOCS3 signaling pathway[J]. Front Biosci (Landmark Ed), 2021, 26:1572–1584.
[34] Li Y, Li H, Chen X, et al.CDK5 regulates PPARgamma/NF-kappaB signaling to exacerbate obesity-
related osteoarthritis via modulating macrophage polarization and chondrocyte apoptosis[J]. Cell Mol Biol Lett, 2025, 30:135.
[35] Zhang LY, Xue HG, Chen JY, et al.Genistein induces adipogenic differentiation in human bone marrow mesenchymal stem cells and suppresses their osteogenic potential by upregulating PPARγ[J]. Exp Ther Med, 2016, 11:1853–1858.
[36] Xu B, Xing A, Li S.The forgotten type 2 diabetes mellitus medicine: rosiglitazone[J]. Diabetol Int, 2021, 13:49–65.
基金
吉林省科技发展计划项目（No.20250601005RC）; 吉林省科技发展计划项目（No.242736NY0102125693）
PDF(11103 KB)
