黄芪甲苷调控JAK2/STAT3/CXCL12信号通路抑制炎症细胞浸润改善小鼠胃炎的机制研究  

Study on the Mechanism of AstragalosideⅣRegulating JAK2/STAT3/CXCL12 Signaling Pathway to Inhibit Inflammatory Cell Infiltration and Improve Gastritis in Mice

作　　者：江晓涛 杨泽虹 王姝烨 安金琪 黄远程 邹雨杉 文艺[2] 李培武[2] JIANG Xiao-Tao;YANG Ze-Hong;WANG Shu-Ye;AN Jin-Qi;HUANG Yuan-Cheng;ZOU Yu-Shan;WEN Yi;LI Pei-Wu(The First Clinical Medical School of Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学第一临床医学院,广东广州510006 [2]广州中医药大学第一附属医院,广东广州510405

出　　处：《广州中医药大学学报》2024年第2期447-456,共10页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：广州中医药大学“双一流”与高水平大学学科协同创新团队项目(编号:2021xk47);广州中医药大学第一附属医院创新强院工程(二期)第二批临床研究专项(编号:2019IIT19)。

摘　　要：【目的】探讨黄芪甲苷对胃炎的抑炎作用及机制。【方法】(1)体外实验:应用1、2.5、5、10、20μg·mL-1脂多糖(LPS)诱导GES-1细胞24 h,确定LPS诱导GES-1细胞炎症模型的最佳浓度。在GES-1细胞炎症模型的基础上,使用0.1、1、10μg·mL-1浓度的黄芪甲苷分别干预24 h,明确黄芪甲苷最佳的干预浓度。采用细胞计数试剂盒8(CCK-8)方法测定黄芪甲苷对细胞活性的影响;采用AutoDock软件对黄芪甲苷和信号传导和转录激活因子3(STAT3)进行分子对接,验证两者的相互作用;采用Western Blot法检测磷酸化STAT3(p-STAT3)和CXCL12蛋白表达水平。(2)体内实验:采用LPS法诱导胃炎小鼠模型,应用黄芪甲苷和p-STAT3抑制剂干预1周后,免疫组织化学法和Western Blot法检测胃黏膜p-STAT3、CXCL12蛋白表达水平,酶联免疫吸附分析(ELISA)检测血浆中CXCL12的含量。【结果】(1)体外实验结果显示:10μg·mL-1是LPS诱导GES-1细胞炎症模型的最佳浓度,选用黄芪甲苷10μg·mL-1开展后续实验。分子对接结果显示,黄芪甲苷与STAT3具有较高的亲和力。与空白对照组比较,LPS诱导组GES-1细胞p-STAT3、CXCL12蛋白表达水平显著升高(P<0.001);黄芪甲苷组和p-STAT3抑制剂组p-STAT3、CXCL12蛋白表达水平较LPS诱导组显著降低(P<0.01)。(2)体内实验结果显示:与空白对照组比较,模型组小鼠胃黏膜p-STAT3、CXCL12蛋白水平和血浆中CXCL12含量显著上调(P<0.01或P<0.001);黄芪甲苷和p-STAT3抑制剂组胃炎小鼠胃黏膜p-STAT3、CXCL12蛋白表达水平和血浆中CXCL12含量显著下调(P<0.05或P<0.01或P<0.001)。【结论】黄芪甲苷通过抑制JAK2/STAT3信号通路降低CXCL12的表达,抑制炎症细胞浸润,达到改善胃炎小鼠胃黏膜炎症的作用。Objective To investigate the anti-inflammatory effect and mechanism of astragalosideⅣon gastritis.Methods(1)In vitro experiments:1,2.5,5,10,20μg·mL-1 lipopolysaccharide(LPS)was applied to induce GES-1 cells for 24 hours,to determine the optimal concentration of LPS for inducing the inflammation model of GES-1 cells.On the basis of the inflammation model of GES-1 cells,astragalosideⅣat concentrations of 0.1,1 and 10μg·mL-1 was used to intervene for 24 hours,respectively,to clarify the optimal concentration of astragalosideⅣfor intervention.The effects of astragalosideⅣon cell activity were determined by the cell counting kit 8(CCK-8)method;molecular docking of astragalosideⅣand signal transducer and activator of transcription 3(STAT3)was performed by using AutoDock software to validate the interaction between the two;and Western Blot was used to detect the phosphorylation of STAT3(p-STAT3)and the protein expression level of CXCL12.(2)In vivo experiments:gastritis was induced in a mouse model by the LPS method,and after 1 week of intervention with astragalosideⅣand protein expression levels of p-STAT3 inhibitor,p-STAT3 and CXCL12 in the gastric mucosa were detected by immunohistochemistry and Western Blot,and CXCL12 content in plasma was detected by enzyme-linked immunosorbent assay(ELISA).Results(1)The results of in vitro experiments showed that 10μg·mL-1 was the optimal concentration for LPS-induced inflammation model of GES-1 cells,and AstragalosideⅣ10μg·mL-1 was chosen to carry out the subsequent experiments.The molecular docking results showed that astragalosideⅣhad a high affinity for STAT3.Compared with the blank control group,the protein expression levels of p-STAT3 and CXCL12 in GES-1 cells in the LPS-induced group were significantly increased(P<0.001);the protein expression levels of p-STAT3 and CXCL12 in the astragalosideⅣgroup and the p-STAT3 inhibitor group were significantly lower than those in the LPS-induced group(P<0.01).(2)The results of in vivo experiments showed that co

关 键 词：黄芪甲苷 胃炎 抑炎作用 STAT3信号通路 CXCL12 GES-1细胞 小鼠 

分 类 号：R285.5[医药卫生—中药学]