通管丸含药血清对巨噬细胞炎症模型磷酸化IRAK4、磷酸化IKKs蛋白表达的影响  

Effect of Tongguan Pills(通管丸)Containing Serum on the Expression of Phosphorylated IRAK4 and Phosphorylated IKKs in Macrophage Inflammatory Model

作　　者：龚倩 匡继林[2] 张翼[2] GONG Qian;KUANG Ji-lin;ZHANG Yi(Hunan University of Chinese Medicine,Changsha Hunan 410208,China;The Second Affiliated Hospital of Hunan University of Chinese Medicine,Changsha Hunan 410005,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]湖南中医药大学第二附属医院,湖南长沙410005

出　　处：《中医药导报》2021年第1期47-50,共4页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：湖南省自然科学基金项目(2017JJ2209),项目名称:基于TLR2/MyD88/NF-κB信号通路的通管丸治疗输卵管炎性阻塞性大鼠的作用机制。

摘　　要：目的:探究通管丸治疗输卵管炎性阻塞性不孕的可能作用机制。方法:将10只大鼠随机分为实验组和对照组。实验组大鼠予通管丸混悬液灌胃,对照组大鼠予等量生理盐水灌胃,连续灌胃3 d后制备血清。将RAW264.7细胞随机分为9组:正常培养组、无药血清组(5%、10%、20%、40%)、含药血清组(5%、10%、20%、40%)。正常培养组添加胎牛血清,其余各组予以相应浓度的血清,CCK8法检测最佳含药血清浓度。将RAW264.7分为造模组和非造模组,造模组添加脂多糖,非造模组添加等量的PBS,ELISA法检测两组TNF-α、IL-1β的含量,确定巨噬细胞炎症模型是否造模成功。最后将RAW264.7分成空白对照组、PPAR-γ激动剂组、NF-κB阻断剂组、无药血清组和含药血清组。空白对照组予以正常RAW264.7细胞和胎牛血清;其余各组予以造模后的RAW264.7细胞和相应的药物。培养后采用Western blotting测定磷酸化IRAK4、IKKs蛋白的表达。结果:与空白对照组比较,无药血清组、NF-κB阻断剂组、PPAR-γ激动剂组、含药血清组中磷酸化IRAK4、IKKs蛋白表达均增加(P<0.05);与无药血清组比较,NF-κB阻断剂组、PPAR-γ激动剂组、含药血清组中磷酸化IRAK4、IKKs蛋白表达均降低(P<0.05)。结论:通管丸可能通过抑制TLR2/MyD88/NF-κB信号通路中"关键元件"磷酸化IRAK4、IKKs蛋白的表达,从而抑制该信号通路的活化,减少炎症因子及其炎症介质的表达,达到抑制输卵管炎症反应的目的。Objective:To explore the possible mechanism of Tongguan Pills in the treatment of tubal inflammatory obstructive infertility.Methods:A total of 10 rats were randomly divided into experimental group and control group.The rats in the experimental group were given the Tongguan Pill suspension by gavage,and the rats in the control group were given the same amount of normal saline by gavage,and serum was prepared after continuous gavage for 3 days.RAW264.7 cells were randomly divided into 9 groups:normal culture group,non-drug serum group(5%,10%,20%,40%),drug-containing serum group(5%,10%,20%,40%).Fetal bovine serum was added to the normal culture group,and the other groups were given corresponding concentrations of serum.The CCK8 method was used to detect the optimal drug-containing serum concentration.RAW264.7 was divided into modeling group and non-modeling group.Lipopolysaccharide(LPS)was added in the modeling group,and PBS was added in the non-modeling group.The levels of TNF-αand IL-1βin both groups were detected by ELISA to determine whether the macrophagic inflammatory model was successfully established.Finally,RAW264.7 was divided into blank control group,PPAR-γagonist group,NF-κB blocker group,non-drug serum group and drug-containing serum group.The blank control group received normal RAW264.7 cells and fetal bovine serum,and the remaining groups received modeled RAW264.7 cells and corresponding drugs.Western blotting was used to detect the expression of phosphorylated IRAK4 and IKKS after culture.Results:Compared with the blank control group,the expression of phosphorylated IRAK4 and IKKs protein increased in non-drug serum group,NF-κB blocker group,PPAR-γagonist group,and drug-containing serum group(P<0.05).Compared with non-drug serum group,the expression of phosphorylated IRAK4 and IKKs protein in the NF-κB blocker group,PPAR-γagonist group,and drug-containing serum group were all decreased(P<0.05).Conclusion:Tongguan Pills may inhibit the expression of IRAK4 and IKKs proteins phosphorylated by"k

关 键 词：输卵管炎性阻塞性不孕 通管丸 磷酸化IRAK4 磷酸化IKKs 信号通路 

分 类 号：R285.5[医药卫生—中药学]