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目的 探索激活α7烟碱型乙酰胆碱受体(α7nAChR)优化肥胖小鼠棕色脂肪组织(BAT)产热作用机制。方法 采用高脂饮食诱导肥胖模型，50只8周龄C57BL/6J小鼠随机分为低脂饮食组(Control组)、高脂饮食组(HFD组)、高脂饮食+β3受体激动剂组(HFD+β3组)、高脂饮食+β3受体激动剂+α7nAChR选择性激动剂组(HFD+β3+GTS-21组)、高脂饮食+β3受体激动剂+α7nAChR选择性抑制剂组(HFD+β3+α-BGT组)。苏木精-伊红(HE)染色观察BAT的形态结构，透射电子显微镜分析脂肪细胞内线粒体数量及脂滴形态变化；ELISA检测BAT中肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-1β、IL-10、转化生长因子β(TGF-β)、环磷酸腺苷(cAMP)以及去甲肾上腺素(NE)水平；RT-qPCR检测BAT中血管内皮生长因子(VEGF-A)、一氧化氮合酶2(NOS2)、精氨酸酶1(Arg1) mRNA的表达水平、以及解偶联蛋白1(UCP1)、含PR结构域蛋白16(PRDM16)、过氧化物酶体增殖物激活受体γ辅激活因子1α(PGC-1α)表达水平；免疫组织化学方法检测棕色脂肪中的CD31、CD86和CD206等巨噬细胞标志物的表达；Western blot检测UCP1、α7nAChR、核因子κB p65(NF-κB p65)、磷酸化Janus激酶2(p-JAK2)和磷酸化信号转导及转录激活因子3(p-STAT3)水平。结果 与Control组比较，HFD组脂肪细胞中脂滴面积增大、线粒体数量减少，促炎因子TNF-α、IL-1β上调，抑炎因子IL-10、TGF-β及Arg1、VEGF-A下调(均P<0.01)。与HFD+β3组相比，HFD+β3+GTS-21组脂肪细胞中脂滴面积减小，线粒体数量增加，促炎因子TNF-α、IL-1β、NOS2及NF-κB p65表达下降，抑炎因子IL-10、TGF-β及Arg1、p-STAT3、pJAK2磷酸化水平升高(均P<0.01)。结论 激活α7nAChR联合β3受体激动剂可有效促进产热反应，改善脂肪组织局部低氧，使局部炎症相关表型向M2样表型偏移，从而缓解BAT炎症状态，增强其产热能力。
Abstract：Objective To investigate the mechanism by which activation of α7nicotinic acetylcholine receptors(α7nAChR) optimizes brown adipose tissue(BAT) thermogenesis in obese mice.Methods Obesity was induced in C57BL/6J mice via a high-fat diet. Fifty 8-week-old mice were randomly assigned to five groups: low-fat diet(Control), high-fat diet(HFD), high-fat diet plus a β3-adrenergic receptor agonist(HFD+β3), high-fat diet plus a β3-adrenergic receptor agonist and an α7nAChR selective agonist(HFD+β3+GTS-21), and high-fat diet plus a β3-adrenergic receptor agonist and an α7nAChR selective antagonist(HFD+β3+α-BGT). Haematoxylin and eosin(HE) staining was used to evaluate BAT morphology. Transmission electron microscopy was performed to assess mitochondrial number and lipid droplet morphology in adipocytes. ELISA was used to measure levels of tumour necrosis factor-α(TNF-α), interleukin(IL)-1β,IL-10, transforming growth factor-β(TGF-β), cyclic adenosine monophosphate(cAMP), and norepinephrine(NE). RT-qPCR was conducted to determine mRNA expression levels of vascular endothelial growth factor A(VEGF-A), nitric oxide synthase 2(NOS2), arginase 1(Arg1), uncoupling protein 1(UCP1), PR domain-containing protein 16(PRDM16), and peroxisome proliferator-activated receptor γ coactivator-1α(PGC-1α) in BAT. Immunohistochemistry was used to detect macrophage markers(CD31, CD86, and CD206). Protein expression levels of UCP1, α7nAChR, nuclear factor-κB p65(NF-κB p65), phosphorylated Janus kinase 2(p-JAK2), and phosphorylated signal transducer and activator of transcription 3(pSTAT3) were analyzed by Western blot.Results Compared with the Control group, the HFD group showed enlarged lipid droplets and reduced mitochondrial numbers in adipocytes, accompanied by increased TNF-α and IL-1β and decreased IL-10, TGF-β, Arg1, and VEGF-A(all P<0. 01). Compared with the HFD+ β3 group, the HFD+β3+GTS-21 group showed a smaller lipid droplet area and a higher mitochondrial number, along with reduced levels of pro-inflammatory factors, NOS2, and NF-κB p65, and increased levels of anti-inflammatory factors, Arg1, and phosphorylation of JAK2 and STAT3(all P<0. 01).Conclusion Activation of α7nAChR in combination with a β3-adrenergic receptor agonist effectively enhances thermogenesis, alleviates local adipose tissue hypoxia, and shifts the local inflammatory phenotype toward an M2-like state, thereby reducing inflammation in brown adipose tissue and improving its thermogenic capacity.
[1]Busebee B, Ghusn W, Cifuentes L, et al. Obesity:a review of pathophysiology and classification[J]. Mayo Clin Proc, 2023, 98(12):1842-57. doi:10. 1016/j. mayocp. 2023. 05. 026.
[2]Harms M, Seale P. Brown and beige fat:development, function and therapeutic potential[J]. Nat Med, 2013, 19(10):1252-63. doi:10. 1038/nm. 3361.
[3]Cheng L, Wang J, Dai H, et al. Brown and beige adipose tissue:a novel therapeutic strategy for obesity and type 2 diabetes mellitus[J]. Adipocyte, 2021, 10(1):48-65. doi:10. 1080/21623945. 2020. 1870060.
[4]Caca J, Bartelt A, Egea V. Hypoxia regulates brown adipocyte differentiation and stimulates miR-210 by HIF-1α[J]. Int J Mol Sci, 2024, 26(1):117. doi:10. 3390/ijms26010117.
[5]Pellegrinelli V, Figueroa-Juárez E, Samuelson I, et al. Defective extracellular matrix remodeling in brown adipose tissue is associated with fibro-inflammation and reduced diet-induced thermogenesis[J]. Cell Rep, 2023, 42(6):112640. doi:10. 1016/j. celrep. 2023. 112640.
[6]Li L, Ma L, Zhao Z, et al. IL-25-induced shifts in macrophage polarization promote development of beige fat and improve metabolic homeostasis in mice[J]. PLoS Biol, 2021, 19(8):e3001348. doi:10. 1371/journal. pbio. 3001348.
[7]马度芳，姜萍，王永成，等.肾上腺素能系统介导的棕色/米色脂肪产热在治疗肥胖中的应用局限性[J].中华内分泌代谢杂志，2017, 33(7):625-8. doi:10. 3760/cma. j. issn. 1000-6699. 2017. 07. 018.[7]Ma D F, Jiang P, Wang Y C, et al. Application limitations of sympathetic nerve-mediated thermogenesis of brown/beige adipose tissue in the treatment of obesity[J]. Chin J Endocrinol Metab,2017, 33(7):625-8. doi:10. 3760/cma. j. issn. 1000-6699. 2017. 07. 018.
[8]Zouali M. Pharmacological and electroceutical targeting of the cholinergic anti-inflammatory pathway in autoimmune diseases[J]. Pharmaceuticals(Basel), 2023, 16(8):1089. doi:10. 3390/ph16081089.
[9]Roa-Vidal N, Rodríguez-Aponte A S, Lasalde-Dominicci J A, et al. Cholinergic polarization of human macrophages[J]. Int J Mol Sci, 2023, 24(21):15732. doi:10. 3390/ijms242115732.
[10]Youssef M E, El-Mas M M, Abdelrazek H M, et al. α7-nAChRsmediated therapeutic angiogenesis accounts for the advantageous effect of low nicotine doses against myocardial infarction in rats[J]. Eur J Pharmacol, 2021, 898:173996. doi:10. 1016/j.ejphar. 2021. 173996.
[11]Buettner R, Scholmerich J, Bollheimer L C. High-fat diets:modeling the metabolic disorders of human obesity in rodents[J].Obes Res, 2007, 15(4):798-808.
[12]Shin W, Okamatsu-Ogura Y, Machida K, et al. Impaired adren--ergic agonistdependent beige adipocyte induction in aged mice[J]. Obesity(Silver Spring), 2017, 25(2):417-23. doi:10. 1002/oby. 21727.
[13]Al-Sharea A, Lee M K S, Whillas A, et al. Nicotinic acetylcholine receptor alpha 7 stimulation dampens splenic myelopoiesis and inhibits atherogenesis in Apoe-/-mice[J]. Atherosclerosis,2017, 265:47-53. doi:10. 1016/j. atherosclerosis. 2017. 08. 010.
[14]Liu Z, Wang Y, Wang Y, et al. Dexmedetomidine attenuates inflammatory reaction in the lung tissues of septic mice by activating cholinergic anti-inflammatory pathway[J]. Int Immunopharmacol, 2016, 35:210-6. doi:10. 1016/j. intimp. 2016. 04. 003.
[15]Becher T, Palanisamy S, Kramer D J, et al. Brown adipose tissue is associated with cardiometabolic health[J]. Nat Med,2021, 27(1):58-65. doi:10. 1038/s41591-020-1126-7.
[16]Della Guardia L, Shin A C. Obesity-induced tissue alterations resist weight loss:a mechanistic review[J]. Diabetes Obes Metab,2024, 26(8):3045-57. doi:10. 1111/dom. 15637.
[17]Zhou Q, Lu Z, Wang B, et al. Endothelial SIRT3 deficiency predisposes brown adipose tissue to whitening in diet-induced obesity[J]. Int J Biol Sci, 2025, 21(8):3444-60. doi:10. 7150/ijbs. 110741.
[18]AlZaim I, de Rooij L P M H, Sheikh B N, et al. The evolving functions of the vasculature in regulating adipose tissue biology in health and obesity[J]. Nat Rev Endocrinol, 2023, 19(12):691-707. doi:10. 1038/s41574-023-00893-6.
[19]Chavakis T, Alexaki V I, Ferrante A W Jr. Macrophage function in adipose tissue homeostasis and metabolic inflammation[J].Nat Immunol, 2023, 24(5):757-66. doi:10. 1038/s41590-023-01479-0.
[20]Pan D, Li G, Jiang C, et al. Regulatory mechanisms of macrophage polarization in adipose tissue[J/OL]. Front Immunol,2023, 14:1149366. doi:10. 3389/fimmu. 2023. 1149366.
[21]Huynh P M, Wang F, An Y A. Hypoxia signaling in the adipose tissue[J]. J Mol Cell Biol, 2025, 16(8):mjae039. doi:10. 1093/jmcb/mjae039.
[22]蒋皓冉，唐晓飞，吴界霖，等. β-石竹烯通过上调PPARγ/PGC-1α/UCP1通路促进肥胖小鼠白色脂肪棕色化作用[J].安徽医科大学学报，2024, 59(9):1591-8. doi:10. 19405/j. cnki.issn1000-1492. 2024. 09. 014[22]Jiang H R, Tang X F, Wu J L, et al. β-caryophyllene promotes white fat browning in obese mice through up-regulation of the PPARγ/PGC-1α/UCP1 pathway[J]. Acta Univ Med Anhui,2024, 59(9):1591-8. doi:10. 19405/j. cnki. issn1000-1492. 2024. 09. 014
[23]Keever K R, Cui K, Casteel J L, et al. Cholinergic signaling via the α7 nicotinic acetylcholine receptor regulates the migration of monocyte-derived macrophages during acute inflammation[J]. J Neuroinflammation, 2024, 21(1):3. doi:10. 1186/s12974-023-03001-7.
[24]吕穆杰，田子芳，王苏童，等.黄芪桂枝五物汤对肥胖小鼠棕色脂肪组织产热的优化作用[J].中华中医药杂志，2023, 38(6):2838-42.[24]Lyu M J, Tian Z F, Wang S T, et al. Optimization of Huangqi Guizhi Wuwu Decoction on thermogenesis of brown adipose tissue in obese mice[J]. China J Tradit Chin Med Pharm, 2023, 38(6):2838-42.
基本信息:
DOI：10.19405/j.cnki.issn1000-1492.2026.07.005
中图分类号:R589.2
引用信息:
[1]田文成,王苏童,范和松,等.激活α7nAChR优化肥胖小鼠棕色脂肪组织产热[J].安徽医科大学学报,2026,61(07):1191-1199.DOI:10.19405/j.cnki.issn1000-1492.2026.07.005.
基金信息:
国家自然科学基金项目(编号：81874449、82274481); 山东省自然科学基金项目(编号：ZR2022LZY004)~~
2026-05-08
2026-05-08
2026-05-08