大黄灵仙方调控IL-2/STAT5信号通路干预Th17/Treg细胞平衡防治肝内胆管炎症  

Mechanism of Dahuang Lingxian(大黄灵仙) Formula in Interfering with Th17/Treg Cell Balance to Prevent and Treat Intrahepatic Bile Duct Inflammation Based on IL-2/STAT5 Signaling Pathway

作　　者：庞浇安 俞渊[1] 胡嗣钦[1] 马天如 叶桂源 常明 李敏鹏 潘孟[1] 陈伟棠 PANG Jiaoan;YU Yuan;HU Siqin;MA Tianru;YE Guiyuan;CHANG Ming;LI Minpeng;PAN Meng;CHEN Weitang(The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530200;Fangchenggang Hospital of Traditional Chinese Medicine,Fangchenggang 538000)

机构地区：[1]广西中医药大学第一附属医院,南宁530200 [2]防城港市中医医院,防城港538000

出　　处：《中药药理与临床》2025年第1期63-70,共8页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金(编号:82360889);广西自然科学基金(编号:2020GXNSFAA238012);2024“岐黄工程”高层次人才团队培育项目(编号:202411);广西中医药大学第一附属医院院级博士启动基金项目(编号:2020BS004);广西中医药大学引进博士科研启动基金项目(编号:2020BS030);基于BOPPPS模式下提高中医外科专业研究生规范化培训教育质量的研究(编号:JGY2020106)。

摘　　要：目的:探讨大黄灵仙方调控IL-2/STAT5信号通路对肝内胆管炎症模型大鼠Th17/Treg细胞平衡的影响。方法:采用随机数字表法将大鼠随机分为正常对照组、模型对照组、胆宁片0.5 g/kg组、大黄灵仙方2.4、4.8 g/kg组,每组10只。造模大鼠在胆总管一次性注射LPS 1.25 mg/kg构建大鼠肝内胆管炎症动物模型。造模成功后各组分别灌胃给予相应浓度药物,正常对照组、模型对照组予等体积生理盐水灌胃,1次/d,连续7 d。HE染色观察肝组织病理变化程度;微板法测定血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBil)、直接胆红素(DBil)的含量;ELISA法检测血清中白介素-2(IL-2)、IL-6、IL-10、IL-17、肿瘤坏死因子-α(TNF-α)的含量;Western blot法检测胆管组织中IL-2、STAT5、p-STAT5蛋白表达;RT-qPCR法检测胆管组织中的Il2、Stat5 mRNA的表达;流式细胞术检测外周血和脾脏CD4、辅助性T细胞17(Th17)、调节性T细胞(Treg)水平,计算Th17/Treg比值。结果:与正常对照组比较,模型对照组大鼠的肝窦、胆管细胞变性增生,胆小管严重淤血扩张,汇管区周围存在大量炎性细胞浸润,血清中ALT、AST、TBil、DBil、IL-6、IL-17、TNF-α活力或含量明显升高,胆管STAT5、p-STAT5蛋白表达上调及Stat5 mRNA表达上调(P<0.01),血清中IL-2、IL-10含量及胆管Il2、Stat5 mRNA表达下调(P<0.01),大鼠外周血和脾脏中CD4、Treg细胞数量减少,Th17细胞数量增加,且Th17/Treg比值显著升高(P<0.01);与模型对照组比较,胆宁片组0.5 g/kg、大黄灵仙方2.4、4.8 g/kg的大鼠肝脏病理损伤程度均有改善,极少见细胞坏死及淤血扩张,血清中ALT、AST、TBil、DBil、IL-6、IL-17、TNF-α活力或含量升高(P<0.05或P<0.01),胆管STAT5、p-STAT5蛋白表达下调,Stat5 mRNA表达下调(P<0.05或P<0.01),Il2、Stat5 mRNA的表达上调,血清中IL-2、IL-10含量升高,大鼠外周血和脾脏中CD4、Treg细胞数量增加,Th17细胞数量减少,且Th17/TregObjective:To explore the effect of Dahuang Lingxian(大黄灵仙) Formula on the helper T cell 17(Th17)/regulatory T cell(Treg) cell balance in a rat model of intrahepatic bile duct inflammation by regulating the IL-2/STAT5 signaling pathway. Methods:The rats were randomly divided into a normal control group, a model control group, a Danning Tablets group(0.5 g/kg),and Dahuang Lingxian Formula groups(2.4 g/kg and 4.8 g/kg) by random number table method, with 10 rats in each group. All model rats were injected with LPS at 1.25 mg/kg into the common bile duct to establish a rat model of intrahepatic bile duct inflammation. After successful modeling, all rats were administered with corresponding drugs by gavage, while rats in the normal control group and model control group were given an equal volume of normal saline by gavage once per day for 7 consecutive days. HE staining was used to observe the degree of pathological changes in liver tissues. The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBil),and direct bilirubin(DBil) in serum were measured by the microplate method. Enzyme-linked immunosorbent assay(ELISA) was used to detect the content of interleukin-2(IL-2),interleukin-6(IL-6),interleukin-10(IL-10),interleukin-17(IL-17),and TNF-α in serum. Western blot and real-time fluorescence quantitative PCR(RT-qPCR) were used to detect the expression levels of IL-2,STAT5 protein, and mRNA in bile duct tissues. Flow cytometry was used to detect the levels of CD4,Th17,and Treg in peripheral blood and spleen, and the Th17/Treg ratio was calculated. Results:Compared with the normal control group, the rats in the model control group showed degeneration and proliferation in liver sinusoids and bile duct cells, severe congestion and dilation of the bile canaliculi, and a large number of inflammatory cell infiltrates around the portal area. The activities or content of ALT,AST,TBil, DBil, IL-6,IL-17,and TNF-α were significantly increased, the protein expression levels of STAT5 and

关 键 词：大黄灵仙方 胆管细胞炎症 白介素-2/信号转导及转录激活因子5 辅助性T细胞17/调节性T细胞 

分 类 号：R73[医药卫生—肿瘤]