摘要
目的基于巨噬细胞极化与半乳糖凝集素3(Gal-3)/Toll样受体4(TLR4)/核因子κB(NF-κB)信号通路探究中药复方汉唐平改善db/db肥胖小鼠脂肪组织慢性炎症的作用机制。方法将40只db/db小鼠随机分为模型组、二甲双胍组(30 mg·kg^(-1))、汉唐平组(50.6 g·kg^(-1))、GB1107组(30 mg·kg^(-1)),每组10只;另外选取10只C57BL/6J小鼠作为对照组;各组均按照10 mL·kg^(-1)给药体积进行灌胃,每日1次,连续12周;对照组、模型组给予等量蒸馏水灌胃。给药期间每2周称量并记录小鼠体质量,取材后称取各组小鼠双侧附睾脂肪组织质量并记录;采用全自动生化仪检测血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)水平;ELISA法检测血清肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、Gal-3水平;HE染色法观察附睾脂肪组织病理变化;免疫组化法检测附睾脂肪组织巨噬细胞标记物分化簇68(CD68)表达情况;免疫荧光法检测附睾脂肪组织巨噬细胞极化标记物分化簇86(CD86)、分化簇206(CD206)表达情况;RT-qPCR法检测附睾脂肪组织促炎因子TNF-α、IL-1β、IL-6及抗炎因子白细胞介素10(IL-10)mRNA表达水平;Western Blot法检测附睾脂肪组织Gal-3/TLR4/NF-κB信号通路相关蛋白的表达水平。结果与对照组比较,模型组小鼠的体质量、附睾脂肪组织质量及血清TC、TG、LDL-C、TNF-α、IL-1β、IL-6、Gal-3水平均显著升高(P<0.001);附睾脂肪细胞大小不一,排列紊乱,质地疏松,细胞横截面积显著增大(P<0.001),且脂肪间质内出现大量呈花冠样结构浸润的炎症细胞;附睾脂肪组织的CD68表达水平显著升高(P<0.001),周围大量巨噬细胞浸润,形成典型花冠样结构;附睾脂肪组织CD86表达水平显著升高(P<0.001),CD206表达水平显著降低(P<0.001),M1型巨噬细胞释放的促炎因子TNF-α、IL-1β、IL-6 mRNA表达水平显著升高(P<0.001),而M2型巨噬细胞释放的抑炎因子IL-10 mRNA表达水平显著降低(P<0.001);附睾脂肪组织Gal-3、TLR4、MyD88蛋白表达水平及IκB-α、NF-κB P65蛋白磷酸化水平均显著升高(P<0.001)。与模型组比较,各给药组小鼠体质量、附睾脂肪组织质量及血清TC、TG、LDL-C、TNF-α、IL-1β、IL-6、Gal-3水平均显著降低(P<0.01,P<0.001);附睾脂肪细胞形态均明显改善,大小趋于均等,排列较为有序,细胞横截面积显著减小(P<0.001),同时花冠样结构浸润的炎症细胞显著减少;附睾脂肪组织的巨噬细胞数量减少,CD68表达水平显著降低(P<0.01,P<0.001);附睾脂肪组织CD86表达水平均显著降低(P<0.001),CD206表达水平显著升高(P<0.01,P<0.001),TNF-α、IL-1β、IL-6 mRNA表达水平显著降低(P<0.01,P<0.001),IL-10 mRNA表达水平显著升高(P<0.05,P<0.01);附睾脂肪组织Gal-3、TLR4、MyD88蛋白表达水平及IκB-α、NF-κB P65蛋白磷酸化水平均显著降低(P<0.05,P<0.01,P<0.001)。结论中药复方汉唐平能够有效减轻db/db肥胖小鼠的体质量及脂肪积聚,纠正脂代谢紊乱,抑制附睾脂肪组织中巨噬细胞浸润和M1型极化,降低促炎因子表达水平,从而改善脂肪组织的慢性炎症反应,其作用机制可能与调控Gal-3/TLR4/NF-κB信号通路的活性有关。
Objective To investigate the mechanism by which the Chinese herbal compound Hantangping(HTP)improves chronic adipose tissue inflammation in db/db obese mice,focusing on macrophage polarization and the Galectin-3(Gal-3)/Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway.Methods Forty db/db mice were randomly divided into four groups(n=10 per group):model,metformin(30 mg·kg^(-1)),HTP(50.6 g·kg^(-1)),and GB1107(30 mg·kg^(-1)),with an additional 10 C57BL/6J mice serving as controls.All groups received intragastric administration(10 mL·kg^(-1))once daily for 12 weeks,while the control and model groups received distilled water.Body mass was measured biweekly.Bilateral epididymal adipose tissue mass was recorded post-harvest.Serum total cholesterol(TC),triglycerides(TG),low-density lipoprotein(LDL-C),TNF-α,IL-1β,IL-6,and Gal-3 levels were analyzed using ELISA.Hematoxylin-eosin(HE)staining assessed adipose tissue histopathology.Immunohistochemistry quantified CD68,CD86,and CD206 expression.RT-qPCR measured TNF-α,IL-1β,IL-6,and IL-10 mRNA levels.Western Blot analyzed Gal-3,TLR4,and NF-κB p65 protein expression.Results Compared with controls,the model group exhibited significantly increased body mass,epididymal adipose tissue mass,and serum TC,TG,LDL-C,TNF-α,IL-1β,IL-6,and Gal-3 levels(P<0.001).Epididymal adipocytes displayed irregular sizes,disorganized arrangement,loose texture,and markedly enlarged cross-sectional areas(P<0.001),with abundant crown-like inflammatory infiltrates.CD68 expression was elevated(P<0.001),indicating extensive macrophage infiltration forming crown-like structures.CD86(M1 marker)expression increased(P<0.001),while CD206(M2 marker)decreased(P<0.001).Pro-inflammatory cytokine(TNF-α,IL-1β,IL-6)mRNA levels rose(P<0.001),whereas anti-inflammatory IL-10 mRNA declined(P<0.001).Gal-3,TLR4,MyD88 protein expression and IκB-α/NF-κB p65 phosphorylation were significantly upregulated(P<0.001).Compared with the model group,all treatment groups showed reduced body mass,epididymal fat mass,and serum TC,TG,LDL-C,TNF-α,IL-1β,IL-6,and Gal-3 levels(P<0.01,P<0.001).Adipocyte morphology improved,with more uniform sizes,organized arrangement,and decreased cross-sectional areas(P<0.001).Crown-like inflammatory infiltrates diminished,along with reduced macrophage numbers and CD68 expression(P<0.01,P<0.001).CD86 expression decreased(P<0.001),while CD206 increased(P<0.01,P<0.001).Pro-inflammatory cytokine mRNA levels declined(P<0.01,P<0.001),whereas IL-10 mRNA increased(P<0.05,P<0.01).Gal-3,TLR4,MyD88 protein expression and IκB-α/NF-κB p65 phosphorylation were suppressed(P<0.05,P<0.01,P<0.001).Conclusion HTP effectively reduces adiposity,corrects dyslipidemia,and mitigates chronic adipose inflammation by suppressing macrophage infiltration and M1 polarization,downregulating pro-inflammatory cytokines,and inhibiting the Gal-3/TLR4/NF-κB pathway.
作者
张钰莹
袁颢瑜
黄薇宇
王保华
李赛美
ZHANG Yuying;YUAN Haoyu;HUANG Weiyu;WANG Baohua;LI Saimei(The First School of Clinical Medicine,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)
出处
《中药新药与临床药理》
北大核心
2025年第10期1636-1645,共10页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金青年基金项目(82104791)
广东省基础与应用基础研究基金面上项目(21202104030001057)。
