基于JAK-STAT信号通路的溃结宁膏对HT-29细胞炎症模型的影响  

Effects of Kuijiening Plaster on Inflammatory Model of HT-29 Cells Based on JAK-STAT Signaling Pathway

作　　者：陈凯 朱莹 CHEN Kai;ZHU Ying(The First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《中国中医药信息杂志》2023年第2期112-117,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(81874466);湖南中医药大学一流学科开放基金(2021ZYX37)。

摘　　要：目的观察溃结宁膏穴位敷贴大鼠血清对人结肠癌HT-29细胞炎症反应和凋亡的影响,基于JAKSTAT信号通路探讨其作用机制。方法将HT-29细胞分为空白组、模型组、空白血清组、溃结宁膏血清组和柳氮磺吡啶(SASP)血清组。采用脂多糖诱导炎症细胞模型,CCK8法检测细胞存活率,TUNEL检测细胞凋亡情况,ELISA检测细胞上清液白细胞介素(IL)-6、IL-10、IL-13含量,RT-PCR检测细胞Janus激酶(JAK)1、JAK2、信号传导及转录激活因子(STAT)3、STAT6基因表达,Western blot检测JAK1、JAK2、STAT3、p-STAT3蛋白表达。结果与空白组比较,模型组和空白血清组细胞存活率明显降低(P<0.05),细胞凋亡率明显升高(P<0.05),细胞上清液IL-6和IL-13含量增加、IL-10含量减少(P<0.05),细胞JAK1、JAK2、STAT3、STAT6基因及JAK1、JAK2、STAT3、p-STAT3蛋白表达明显升高(P<0.05);与模型组及空白血清组比较,溃结宁膏血清组和SASP血清组细胞存活率明显增加(P<0.05),细胞凋亡率明显降低(P<0.05),细胞上清液IL-6和IL-13含量减少、IL-10含量增加(P<0.05),细胞JAK1、JAK2、STAT3、STAT6基因及JAK1、JAK2、STAT3、p-STAT3蛋白表达明显降低(P<0.05)。结论溃结宁膏穴位敷贴大鼠血清可能通过调控JAK-STAT信号通路调节HT-29细胞炎症反应及凋亡。Objective To observe the effects of acupoint application rat serum of Kuijiening Plaster on inflammatory response and apoptosis of HT-29 cells;To discuss its mechanism of action based on JAK-STAT signaling pathway.Methods HT-29 cells were divided into blank group,model group,blank serum group,Kuijiening Plaster serum group,and sulfadiazine(SASP)serum group.Inflammatory cell model was induced by LPS.CCK8 method was used to detect cell viability,TUNEL staining was used to detect the apoptosis,ELISA was used to detect the contents of IL-6,IL-10 and IL-13 in cell culture medium,RT-PCR was used to detect cellular Janus kinase(JAK)1,JAK2,signal transducer and activator of transcription(STAT)3,STAT6 gene expression,Western blot was used to detect JAK1,JAK2,STAT3 and p-STAT3 protein expression.Results Compared with the blank group,the cell survival rate of model group and blank serum group significantly decreased(P<0.05),the apoptosis rate significantly increased(P<0.05),the contents of IL-6 and IL-13 in supernatant increased,the content of IL-10 reduced(P<0.05),and the expressions of JAK1,JAK2,STAT3 and STAT6 gene and JAK1,JAK2,STAT3 and p-STAT3 protein in cells significantly increased(P<0.05).Compared with the model group and the blank serum group,the cell survival rate in Kuijiening Plaster serum group and SASP serum group significantly increased(P<0.05),the apoptosis rate significantly decreased(P<0.05),the contents of IL-6 and IL-13 in cell supernatant reduced,the content of IL-10 increased(P<0.05),and the expressions of JAK1,JAK2,STAT3 and STAT6 gene and JAK1,JAK2,STAT3 and p-STAT3 protein in cells significantly decreased(P<0.05).Conclusion Acupoint application rat serum of Kuijiening Plaster may regulate the inflammatory response and apoptosis of HT-29 cells by regulating JAK-STAT signaling pathway.

关 键 词：溃疡性结肠炎 溃结宁膏 JAK-STAT信号通路 HT-29细胞 

分 类 号：R285.5[医药卫生—中药学]