黄芪-败酱草药对对HT-29细胞炎症模型JAK1/STAT6/SOCS1信号通路的影响  

Effect of Drug Pair Huangqi(Radix Astragali)&Baijiangcao(Herba Patriniae)on HT-29 Cell Inflammation Model by Regulating JAK1/STAT6/SOCS1 Signaling Pathway

作　　者：孙大娟[1] 魏秀楠 程艳[1] 迟莉丽[1] SUN Dajuan;WEI Xiunan;CHENG Yan;CHI Lili(The Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan,250014;Shandong University of Traditional Chinese Medicine)

机构地区：[1]山东中医药大学附属医院,山东省济南市250014 [2]山东中医药大学

出　　处：《中医杂志》2023年第3期295-302,共8页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81673969,82205057)。

摘　　要：目的探讨黄芪-败酱草药对治疗溃疡性结肠炎(UC)的可能作用机制。方法脂多糖(LPS)20μg/ml干预HT-29细胞24 h,诱导建立炎症细胞模型。采用8个不同浓度(1.125、2.25、4.5、9、18、36、72、144 mg/ml)黄芪-败酱草干预炎症细胞模型24 h,CCK-8法检测细胞活力,选取细胞活力最高的3个黄芪-败酱草浓度用于后续实验。实验分为空白组、模型组、美沙拉嗪组及黄芪-败酱草低、中、高剂量组6组。除空白组外,其余各组细胞建立炎症模型后,美沙拉嗪组加入1 mg/ml的5-氨基水杨酸100μl,黄芪-败酱草低、中、高剂量组分别加入筛选浓度的低、中、高浓度黄芪-败酱草100μl,空白组、模型组均加入100μl完全培养基,干预24 h。ELISA法检测炎性因子白细胞介素13(IL-13)、白细胞介素6(IL-6)、白细胞介素18(IL-18)、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)表达;Western blot法检测咬合蛋白(Occludin)、紧密连接蛋白1(Claudin-1)、磷酸化蛋白质酪氨酸激酶1(p-JAK1)、磷酸化信号转导和转录激活因子6(p-STAT6)、蛋白质酪氨酸激酶1(JAK1)、信号转导和转录激活因子6(STAT6)、细胞因子信号传导抑制蛋白1(SOCS1)蛋白表达,qRT-PCR法检测JAK1、STAT6、SOCS1 mRNA表达。结果最终选择黄芪-败酱草浓度2.25、4.5、9 mg/ml作为低、中、高浓度进行后续实验。与模型组比较,黄芪-败酱草高剂量组和美沙拉嗪组IL-6、IL-1β、IL-18、TNF-α表达减少,IL-13表达增多,Occludin、Claudin-1、SOCS1蛋白表达及SOCS1 mRNA表达上调,p-JAK1、p-STAT6蛋白表达及JAK1、STAT6 mRNA表达下调(P<0.05或P<0.01)。与美沙拉嗪组比较,黄芪-败酱草高剂量组IL-6、IL-1β、TNF-α升高,IL-13表达降低(P<0.05或P<0.01),Occludin、Claudin-1蛋白表达,JAK1、STAT6、SOCS1 mRNA表达差异均无统计学意义(P>0.05)。结论黄芪-败酱草药对可能通过JAK1/STAT6/SOCS1通路下调致炎因子表达,从而发挥抗炎、修复溃疡性结肠炎肠黏�Objective To explore the possible mechanism of drug pair Huangqi(Radix Astragali)&Baijiangcao(Herba Patriniae)in the treatment of ulcerative colitis(UC).Methods After 24-hour intervention of 20μg/ml lipopolysaccharide(LPS)on HT-29 cells,an inflammatory cell model was established.Huangqi&Baijiangcao at eight different concentrations(1.125,2.25,4.5,9,18,36,72,144 mg/ml)were used on the inflammatory cell model for 24 hours,and the cell viability was detected by CCK-8 method to obtain the top three for follow-up experiments.There were six groups including blank group,model group,mesalazine group and Huangqi&Baijiangcao low-,middle-and high-dose groups.After modeling,100μl of 1 mg/ml 5-aminosalicylic acid was given to the mesalazine group,and 100μl of the obtained low,medium and high concentrations of Huangqi&Baijiangcao was given to the corresponding three Huangqi-Baijiangcao groups,respectively;100μl complete medium was used in the blank group and model group.All groups were intervened for 24 hours.The expression of inflammatory factors interleukin 13(IL-13),interleukin 6(IL-6),interleukin 18(IL-18),tumor necrosis factorα(TNF-α)and interleukin 1β(IL-1β)was detected by ELISA.The protein expression of Occludin,Claudin-1,phosphorylated protein tyrosine kinase 1(p-JAK1),phosphorylated signal transducer and activator of transcription 6(p-STAT6),protein tyrosine kinase 1(JAK1),signal transducer and activator of transcription 6(STAT6),and suppressor of cytokine signaling 1(SOCS1)was detected by Western blot,and the mRNA expression of JAK1,STAT6,SOCS1 was detected by qRT-PCR.Results Finally,2.25 mg/ml,4.5 mg/ml and 9 mg/ml of Huangqi&Baijiangcao were obtained as low,medium and high concentrations.Compared to those in the model group,the expressions of IL-6,IL-1β,IL-18 and TNF-αdecreased in high-dose Huangqi&Baijiangcao group and mesalamine groups,while the expression of IL-13 increased;the protein expression of Occludin,Claudin-1 and SOCS1,as well as the SOCS1 mRNA expression were up-regulated,while p-JAK1,p-STAT6

关 键 词：溃疡性结肠炎 黄芪 败酱草 炎性因子 肠黏膜屏障 JAK1/STAT6/SOCS1通路 

分 类 号：R285[医药卫生—中药学]