摘要
目的:观察前列消汤干预转化生长因子-β1(TGF-β1)-Smad同源物2/3(Smad2/3)信号通路调控自身免疫性前列腺炎(EAP)辅助性T细胞17(Th17)分化机制。方法:120只无特定病原体(SPF)级雄性C57BL/6小鼠随机分为空白组、模型组、阳性对照(吡非尼酮)组、前列消汤低/中/高剂量组,每组20只,采用免疫佐剂结合中医湿热证候因素诱导建立EAP小鼠湿热证模型,造模结束后给予相应药物处理;苏木精-伊红(HE)染色观察前列腺、脾脏组织病理改变,实时荧光定量逆转录聚合酶链反应(RT-PCR)与蛋白质印迹法(Western Blotting)检测前列腺组织中TGF-β1、TGF-β1受体Ⅰ(TGF-β1RⅠ)、白细胞介素17-A(IL-17A)、核转录因子维甲酸相关孤核受体γt(RORγt)、Smad2磷酸化(p-Smad2)、Smad2、p-Smad3和Smad3 mRNA及蛋白表达水平。结果:与空白组比较,模型组小鼠前列腺指数和湿热证候积分明显增高,前列腺和脾脏组织出现大量炎症细胞浸润,提示EAP小鼠湿热证模型制作成功,TGF-β1、TGF-β1RⅠ、IL-17A、RORγt、p-Smad2和p-Smad3表达显著增高(P<0.05);与模型组比较,吡非尼酮组及前列消汤低/中/高剂量组均能不同程度改善炎症浸润程度,TGF-β1、TGF-β1RⅠ、IL-17A、RORγt、p-Smad2和p-Smad3表达显著降低(P<0.05)。结论:前列消汤能有效抑制自身免疫性前列腺炎Th17细胞异常分化功能,其作用机制可能与TGF-β1-Smad2/3信号通路相关。
Objective:To observe the effect of Qianliexiao Decoction on the regulation of the transforming growth factor-β1(TGF-β1)-Smad homolog 2/3(Smad2/3)signaling pathway in the differentiation of Th17 cells in experimental autoimmune prostatitis(EAP).Methods:A total of 120 specific-pathogen-free(SPF)male C57BL/6 mice were randomly divided into six groups:blank group,model group,positive control(pirfenidone)group,and low,medium,and high dose Qianliexiao Decoction groups,with 20 mice in each group.An EAP mouse model with dampness-heat syndrome was established through immune adjuvant combined with traditional Chinese medicine(TCM)dampness-heat factors.After model establishment,the corresponding treatments were administered.Hematoxylin-eosin(HE)staining was used to observe pathological changes in the prostate and spleen tissues.Real-time quantitative reverse transcription polymerase chain reaction(RT-PCR)and Western Blotting(WB)analysis were used to detect the expression levels of TGF-β1,TGF-β1 receptor I(TGF-β1RⅠ),interleukin 17-A(IL-17A),retinoid-related orphan receptorγt(RORγt),phosphorylated Smad2(p-Smad2),Smad2,phosphorylated Smad3(p-Smad3),and Smad3 mRNA and protein in prostate tissue.Results:Compared with the blank group,the model group showed significantly increased prostate index and dampness-heat syndrome scores,and extensive infiltration of inflammatory cells in the prostate and spleen tissues,indicating successful establishment of the EAP model in mice with dampness-heat syndrome.The expression levels of TGF-β1,TGF-β1RⅠ,IL-17A,RORγt,p-Smad2,and p-Smad3 were significantly increased(P<0.05).Compared with the model group,both the pirfenidone group and all doses of the Qianliexiao Decoction groups showed varying degrees of improvement in inflammatory infiltration,with significantly reduced expression of TGF-β1,TGF-β1RⅠ,IL-17A,RORγt,p-Smad2,and p-Smad3(P<0.05).Conclusion:Qianliexiao Decoction can effectively inhibit the abnormal differentiation of Th17 cells in EAP,and its mechanism of action may be related to the TGF-β1-Smad2/3 signaling pathway.
作者
张燕华
买鹏宇
朱闽
叶斌
张泽朝
ZHANG Yanhua;MAI Pengyu;ZHU Min;YE Bin;ZHANG Zechao(Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530011,China;Graduate School of Guangxi University of Chinese Medicine,Nanning 530001,China)
出处
《世界中医药》
北大核心
2024年第22期3437-3443,3451,共8页
World Chinese Medicine
基金
国家自然科学基金项目(81660796,81860847)
广西自然科学基金项目(2023GXNSDA026014)
广西自然科学基金青年科学基金项目(2021GXNSFBA196078)
广西中医药大学“桂派杏林拔尖人才”项目(2022C024)
广西研究生教育创新计划项目(YCSZ2022020)。
