探讨植物乳植杆菌(Lactiplantibacillus plantarum, LP)对Th9细胞分化的调控作用及其在糖尿病相关甲状腺疾病中的潜在免疫调节作用,并初步探索其可能涉及的STING-NLRP3信号通路。分离小鼠脾脏CD4+初始T细胞,用TGF-β1和IL-4诱导分化为Th9细胞,设立对照组和LP干预组,培养48 h。采用流式细胞术检测Th9细胞比例;通过RT-PCR和Western Blot检测PU.1、IL-9、STING、NLRP3及其下游分子的mRNA和蛋白表达。构建2型糖尿病(type 2 diabetes mellitus,T2DM)和甲状腺结节(thyroid nodule,TN)小鼠模型,给予LP干预,检测血清炎症因子水平、Th9细胞比例以及STING-NLRP3信号通路相关分子表达变化。LP干预可显著抑制体外诱导的Th9细胞中IL-9的表达(P<0.01),同时降低STING和NLRP3蛋白表达水平(P<0.01)。LP通过下调STING表达,抑制NLRP3炎症小体的活化,进而减少IL-1β的产生,最终影响Th9细胞分化及IL-9的分泌。在T2DM合并TN小鼠模型中,LP干预可降低血清中促炎因子水平,减少脾脏组织中Th9细胞比例(P<0.01),并通过调节STING-NLRP3信号轴减轻甲状腺组织炎症反应。L. plantarum可抑制Th9细胞分化及功能,减轻糖尿病相关甲状腺疾病的炎症反应,这一作用可能与STING-NLRP3信号通路的调节有关,为相关疾病的防治提供新的潜在靶点。
To investigate the regulatory effects of Lactiplantibacillus plantarum (LP) on Th9 cell differentiation and its potential immunomodulatory role in diabetes-related thyroid diseases, and to preliminarily explore the potentially involved STING-NLRP3 signaling pathway.CD4+ naive T cells in the spleen of mice were isolated and differentiated into Th9 cells induced by TGF-β1 and IL-4, and the control group and LP intervention group were set up and cultured for 48 hours.Flow cytometry was used to detect the proportion of Th9 cells.The mRNA and protein expressions of PU.1, IL-9, STING, NLRP3 and their downstream molecules were detected by RT-PCR and Western blot.Mouse models of type 2 diabetes mellitus (T2DM) and thyroid nodules (TN) were constructed, and LP intervention was given to detect the changes in serum inflammatory factor levels, the proportion of Th9 cells and the expression of molecules related to STING-NLRP3 signaling pathway.LP intervention significantly inhibited the expression of IL-9 in Th9 cells induced in vitro (P<0.01), and reduced the protein expression levels of STING and NLRP3 (P<0.01).LP inhibits the activation of NLRP3 inflammasome by down-regulating STING expression, thereby reducing the production of IL-1β, and ultimately affecting Th9 cell differentiation and IL-9 secretion.In a mouse model of T2DM combined with TN, LP intervention reduced the level of pro-inflammatory factors in serum, reduced the proportion of Th9 cells in spleen tissues (P<0.01), and alleviated the inflammatory response of thyroid tissue by regulating the STING-NLRP3 signaling axis.Lactobacillus plantarum can inhibit the differentiation and function of Th9 cells, alleviate the inflammatory response in diabetes-associated thyroid diseases.This effect may be related to the regulation of the STING-NLRP3 signaling pathway, providing new potential targets for the prevention and treatment of related diseases.
[1] TOULIS K, TSEKMEKIDOU X, POTOLIDIS E, et al.Thyroid autoimmunity in the context of type 2 diabetes mellitus:Implications for vitamin D[J].International Journal of Endocrinology, 2015, 2015:710363.
[2] ZHENG Y, LEY S H, HU F B.Global aetiology and epidemiology of type 2 diabetes mellitus and its complications[J].Nature Reviews Endocrinology, 2018, 14(2):88-98.
[3] WANG X, BAO W, LIU J, et al.Inflammatory markers and risk of type 2 diabetes:A systematic review and meta-analysis[J].Diabetes Care, 2013, 36(1):166-175.
[4] FERRARI S M, FALLAHI P, GALDIERO M R, et al.Immune and inflammatory cells in thyroid cancer microenvironment[J].International Journal of Molecular Sciences, 2019, 20(18):4413.
[5] SWANSON K V, DENG M, TING J P Y.The NLRP3 inflammasome:Molecular activation and regulation to therapeutics[J].Nature Reviews Immunology, 2019, 19(8):477-489.
[6] REN W X, SUN Y, ZHAO L, et al.NLRP3 inflammasome and its role in autoimmune diseases:A promising therapeutic target[J].Biomedicine & Pharmacotherapy, 2024, 175:116679.
[7] KELLEY N, JELTEMA D, DUAN Y H, et al.The NLRP3 inflammasome:An overview of mechanisms of activation and regulation[J].International Journal of Molecular Sciences, 2019, 20(13):3328.
[8] ZHANG Z L, ZHOU H F, OUYANG X H, et al.Multifaceted functions of STING in human health and disease:From molecular mechanism to targeted strategy[J].Signal Transduction and Targeted Therapy, 2022, 7:394.
[9] HE X, WEDN A, WANG J, et al.IUPHAR ECR review:The cGAS-STING pathway:Novel functions beyond innate immune and emerging therapeutic opportunities[J].Pharmacological Research, 2024, 201:107063.
[10] LI N, ZHOU H, WU H M, et al.STING-IRF3 contributes to lipopolysaccharide-induced cardiac dysfunction, inflammation, apoptosis and pyroptosis by activating NLRP3[J].Redox Biology, 2019, 24:101215.
[11] ZHONG W Z, RAO Z Q, RAO J H, et al.Aging aggravated liver ischemia and reperfusion injury by promoting STING-mediated NLRP3 activation in macrophages[J].Aging Cell, 2020, 19(8):e13186.
[12] LI J H, CHEN S Q, XIAO X, et al.IL-9 and Th9 cells in health and diseases:From tolerance to immunopathology[J].Cytokine & Growth Factor Reviews, 2017, 37:47-55.
[13] ZHANG W C, TANG T T, NIE D A, et al.IL-9 aggravates the development of atherosclerosis in ApoE-/-mice[J].Cardiovascular Research, 2015, 106(3):453-464.
[14] MAANIITTY E, SINISILTA S, JALKANEN J, et al.Distinct circulating cytokine levels in patients with angiography-proven coronary artery disease compared to disease-free controls[J].International Journal of Cardiology Cardiovascular Risk and Prevention, 2024, 22:200307.
[15] SIMONOVIC S Z, MIHALJEVIC O, MAJSTOROVIC I, et al.Cytokine production in peripheral blood cells of patients with differentiated thyroid cancer:Elevated Th2/Th9 cytokine production before and reduced Th2 cytokine production after radioactive iodine therapy[J].Cancer Immunology, Immunotherapy, 2015, 64(1):75-82.
[16] WANG J, JI H F, WANG S X, et al.Probiotic Lactobacillus plantarum promotes intestinal barrier function by strengthening the epithelium and modulating gut microbiota[J].Frontiers in Microbiology, 2018, 9:1953.
[17] ZENG Z B, HUANG Z H, YUE W, et al.Lactobacillus plantarum modulate gut microbiota and intestinal immunity in cyclophosphamide-treated mice model[J].Biomedicine & Pharmacotherapy, 2023, 169:115812.
[18] XIA Y J, CHEN Y, WANG G Q, et al.Lactobacillus plantarum AR113 alleviates DSS-induced colitis by regulating the TLR4/MyD88/NF-κB pathway and gut microbiota composition[J].Journal of Functional Foods, 2020, 67:103854.
[19] SPECTOR L, SUBRAMANIAN N.Revealing the dance of NLRP3:Spatiotemporal patterns in inflammasome activation[J].Immunometabolism, 2025, 7(1):e00053.
[20] LI W Y, SHEN N, KONG L Q, et al.STING mediates microglial pyroptosis via interaction with NLRP3 in cerebral ischaemic stroke[J].Stroke and Vascular Neurology, 2024, 9(2):153-164.
[21] CHEN J, GUAN L, TANG L, et al.T helper 9 cells:A new player in immune-related diseases[J].DNA and Cell Biology, 2019, 38(10):1040-1047.
[22] VYAS S P, GOSWAMI R.A decade of Th9 cells:Role of Th9 cells in inflammatory bowel disease[J].Frontiers in Immunology, 2018, 9:1139.
[23] MOHAMMED HUSSEIN S M, ABDELMAGEED R M.The relationship between type 2 diabetes mellitus and related thyroid diseases[J].Cureus, 2021, 13(12):e20697.
[24] EOM Y S, WILSON J R, BERNET V J.Links between thyroid disorders and glucose homeostasis[J].Diabetes & Metabolism Journal, 2022, 46(2):239-256.
[25] YANG B, YUE Y, CHEN Y, et al.Lactobacillus plantarum CCFM1143 alleviates chronic diarrhea via inflammation regulation and gut microbiota modulation:A double-blind, randomized, placebo-controlled study[J].Frontiers in Immunology, 2021, 12:746585.
[26] WU Y Q, LI A, LIU H W, et al.Lactobacillus plantarum HNU082 alleviates dextran sulfate sodium-induced ulcerative colitis in mice through regulating gut microbiome[J].Food & Function, 2022, 13(19):10171-10185.
[27] KIM H I, KIM J K, KIM J Y, et al.Lactobacillus plantarum LC27 and Bifidobacterium longum LC67 simultaneously alleviate high-fat diet-induced colitis, endotoxemia, liver steatosis, and obesity in mice[J].Nutrition Research, 2019, 67:78-89.
[28] ZHENG D P, LIWINSKI T, ELINAV E.Interaction between microbiota and immunity in health and disease[J].Cell Research, 2020, 30(6):492-506.
