溃结宁膏调控microRNA-155介导JAK/STAT通路对HT-29细胞炎症模型的影响及作用机制研究  

Mechanism of Kuijiening Plaster on inflammation model of HT-29 cells by regulating microRNA-155 mediated JAK/STAT pathway

作　　者：袁宇晴 娄余 龙丹 徐寅[1] 朱莹 YUAN Yuqing;LOU Yu;LONG Dan;XU Yin;ZHU Ying(Department of Spleen and Stomach Diseases,the First Hospital of Hunan University of Chinese Medicine,Changsha,410021,China;Department of Spleen and Stomach Diseases,the First Affiliated Hospital of Guangxi University of Chinese Medicine)

机构地区：[1]湖南中医药大学第一附属医院脾胃病科,长沙410021 [2]广西中医药大学第一附属医院脾胃病科

出　　处：《中国中西医结合消化杂志》2023年第4期285-292,共8页Chinese Journal of Integrated Traditional and Western Medicine on Digestion

基　　金：国家自然科学基金项目(No:81874466);湖南省自然科学基金面上项目(No:2021JJ30531);湖南省教育厅科学研究项目重点项目(No:20A378)。

摘　　要：目的:研究溃结宁膏对HT-29细胞炎症模型的影响并基于microRNA-155(miR-155)/酪氨酸蛋白激酶(janus kinase,JAK)/信号转导及转录激活因子(signal transducer and activator of transcription,STAT)通路探讨其机制。方法:体外培养HT-29细胞建立溃疡性结肠炎炎症模型,分为空白对照组、炎症模型组、模型对照组、miR-155 mimics组、miR-155 inhibitor组、穴位敷贴组、柳氮磺吡啶(SASP)组。乳酸脱氢酶法检测细胞损伤,ELISA检测细胞培养液IL-8、IL-6的含量。qRT-PCR检测miR-155、JAK2、STAT3 mRNA的表达,免疫荧光检测细胞因子信号传导抑制蛋白1(suppressor of cytokine signaling,SOCS1)、JAK2/p-JAK2、STAT3/p-STAT3蛋白的表达水平。结果:与空白对照组比较,模型组和模型对照组细胞乳酸脱氢酶释放率,IL-6、IL-8含量,miR-155、JAK2/p-JAK2、STAT3/p-STAT3 mRNA和蛋白表达量明显增加,差异有统计学意义(P<0.05)。与模型对照组比较,穴位敷贴组细胞乳酸脱氢酶释放率,IL-6、IL-8含量,JAK2/p-JAK2,STAT3/p-STAT3 mRNA和蛋白表达显著减少,miR-155 mRNA表达量显著降低,SOCS1蛋白表达显著增加,差异有统计学意义(P<0.05)。与miR-155 mimics组比较,miR-155 inhibitor组细胞中JAK2/p-JAK2、p-STAT3蛋白的表达显著降低,SOCS1蛋白的表达明显增加,差异有统计学意义(P<0.05)。结论:溃结宁膏穴位敷贴可能通过调控miR-155/JAK/STAT信号通路调节HT-29细胞炎症反应。Objective:To investigate the effect of Kuijiening Plaster on the inflammation model of HT-29 cells and explore the mechanism based on microRNA-155(miR-155)/janus kinase(JAK)/signal transducer and activator of transcription(STAT)pathway.Methods:HT-29 cells were cultured in vitro to establish an inflammatory model of ulcerative colitis.The cells were divided into a blank control group,an inflammation model group,the model control group,the miR-155 mimics group,the miR-155 inhibitor group,the acupuncture point dressing group,and the salazosulfapyridine(SASP)group.The release of lactate dehydrogenase(LDH)was used to detect cell damage,and ELISA was used to detect the content of interleukin IL-8 and IL-6.The mRNA expression of miR-155,JAK2,STAT3 was detected by qRT-PCR,and protein expression of suppressor of cytokine signaling 1(SOCS1),JAK2/p-JAK2,STAT3/p-STAT3 was detected by immunofluorescence.Results:Compared with the blank control group,the LDH release rate,IL-6 and IL-8 contents,the mRNA and protein expression of miR-155,JAK2/p-JAK2,STAT3/p-STAT3 in the model and model control groups were significantly increased(P<0.05).Compared with the model control group,the cell LDH release rate,content of IL-6,IL-8,mRNA and protein expression of JAK2/p-JAK2,STAT3/p-STAT3 were significantly decreased,mRNA expression of miR-155 was significantly decreased,and protein expression of SOCSl was significantly increased in the acupressure group(P<0.05).Compared with the miR-155 mimics group,the protein expression of JAK2/p-JAK2 and p-STAT3 was significantly decreased and the protein expression of SOCS1 was significantly increased in the miR-155 inhibitor group(P<o.05).Conclusion:Kuijiening Plaster acupoint application serum may regulate the inflammatory response of HT-29 cells by regulating miR-155/JAK/STAT signal pathway.

关 键 词：溃疡性结肠炎 溃结宁膏 microRNA-155 酪氨酸蛋白激酶 信号转导及转录激活因子 HT-29细胞 炎症 

分 类 号：R574.62[医药卫生—消化系统]