对黄芪调节革兰阳性菌脓毒症炎症反应部分机制的探讨  

The partial mechanism by which astragalus regulates the inflammatory response in gram - positive bacterial sepsis

作　　者：路玲[1] 王勇强[1] 高红梅[1] 

机构地区：[1]天津市第一中心医院ICU,天津300192

出　　处：《中国急救医学》2015年第1期9-14,共6页Chinese Journal of Critical Care Medicine

基　　金：天津市卫生局攻关课题（2010KG103）;国家临床重点专科建设经费资助（天津市第一中心医院重症医学科）

摘　　要：目的观察脂磷壁酸（LTA）对单核细胞系（U937）刺激后中药黄芪对其所表达的炎性因子的影响，探讨其调节革兰阳性菌脓毒症炎症反应的部分机制。方法体外培养U937细胞，选取不同浓度LIA进行孵育，在不同时间点进行相关测定。MTr法检测细胞的增殖活性，酶联免疫吸附测定法（ELISA法）检测上清液中促炎因子IL-8及抗炎因子IL-10的表达，实时荧光定量聚合酶链反应（RT—PCR）检测细胞核内IL-8mRNA及IL-10mRNA的表达。结果LTA（3—30mg／mL）刺激U937细胞32h内，细胞增殖活性无明显变化（P〉0．05）；而以LTA60mg／mL组及3～30mg／mLLTA刺激40h时，细胞增殖活性明显降低（P〈0．05）。ELISA结果显示，LTA能明显上调U937表达促炎因子IL-8及抗炎因子IL-10，且其表达与LTA浓度及时间呈正相关，其中LTA30mg／mL在8～32h时IL-8及IL-10表达有统计学意义（尸〈0．05），且以16h时IL-8的释放量为最高峰，以24h时IL—10的释放量为最高峰。黄芪可减低LTA刺激U937表达促炎因子IL-8含量，促进LTA刺激U937表达抗炎因子IL-10的释放量，且黄芪0．2mg／mL组在16h、24h及32h时与LTA30组比较差异有统计学意义（P〈0．05），IL-10的释放高峰提前至16h。RT—PCR结果显示，IJTA组和LTA＋黄芪组IL-8mRNA、IL-10mRNA表达及变化趋势与IL-8和IL-10一致。结论中药黄芪在革兰阳性菌脓毒症炎症反应中从细胞水平可抑制促炎因子基因及蛋白的表达，同时可促进细胞水平抗炎因子基因及蛋白的表达，并可使抗炎因子的释放高峰提前。Objective To investigate the expression of inflammatory factors in the U937 cells induced by lipoteichoic acid （LTA）, which was influenced by astragalus membranaceus. Methods The U937 cells was cultured in vitro with LTA. MTY assay was used to detect ceils proliferation activity. The expression of IL- 8 and IL- 10 was observed with enzyme -linked immunosorbent test （ELISA）. The expression of IL- 8 mRNA and IL- 10 mRNA were evaluated by Fluorescence Detection Quantitative Polymerase Chain Reaction （PCR）. Results U937 cells stimulated by LTA, the density between 3 mg/mL and 30 mg/mL within 32 h, did not shown apparent changes by MTT assay （P 〉 0. 05 ） ; but with 60 mg/mL LTA stimulated the cells, the number of cells reduced significantly （ P 〈 0. 05）. LTA can obviously increase U937 expressing pro -inflammatory factor IL- 8 and anti - inflammatory factor IL - 10. And with 30 mg/mL LTA stimulation, the expression of IL - 8 and IL - 10 was significantly increased in 8 h, 16 h, 24 h and 32 h, release of IL - 8 to peak in 16 h, release of IL - 10 for the highest peak in 24 h. Astragalus membranaceus could reduce the LTA to stimulate U937 expressing ｜L -8, can promote LTA to stimulate U937 expressing IL - 10, the peak of the release of IL-10ahead of time to 16 h. The expression of IL-8 mRNA and IL-10 mRNA fit in with the changing of IL - 8 and IL - 10. Conclusion Chinese traditional medicine astragalus membranaceus can inhibit cellular level pro - inflammatory factor gene and protein expression, at the same time can promote cell level anti - inflammatory factor gene and protein expression, and can make the anti - inflammatory factors release peak in advance.

关 键 词：脂磷壁酸(LTA) U937细胞系 促炎因子 抗炎因子 

分 类 号：R978.1[医药卫生—药品]