芪参合剂对耐药肺炎克雷伯菌脓毒症大鼠的固有免疫调节作用研究

A Study on the Regulating Action of Qishen Mixture for Innate Immunity of Drug-resistant Pneumoniae Klebsiella Sepsis Rats

目的:研究芪参合剂对耐药肺炎克雷伯菌脓毒症大鼠的固有免疫调节作用。方法:将50只SD大鼠随机分成正常组、模型组及芪参合剂低、中、高剂量组,每组10只。耐药肺炎克雷伯菌气管注射复制脓毒症大鼠模型,造模后6小时按分组灌胃方式给予相对应剂量芪参合剂或生理盐水,干预3天后取支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)进行白细胞总数及白细胞分类计数;取支气管肺泡灌洗液(BALF)、血液(blood)、脾脏(spleen)、肾脏(kidney)进行菌落形成单位(colony-forming units,CFU)测定;ELISA法检测BALF中肿瘤坏死因子α(tumor necrosis factorα,TNF-α)、白细胞介素6(interleukin-6,IL-6)和肺组织中CXC趋化因子配体(C-X-C motif chemokine ligand-1,CXCL1)、巨噬细胞炎症因子蛋白2(macrophage inflammatory protein 2,MIP-2)表达。结果:模型组大鼠BALF中白细胞总数及白细胞分类计数明显升高,BALF、blood、脾、肾组织中细菌负荷明显增加,且组织细胞因子TNF-α、IL-6、CXCL1、MIP-2表达明显升高。3种剂量芪参合剂干预后,能不同程度降低BALF中白细胞总数及白细胞分类计数,减轻主要脏器细菌负荷,且可以明显降低细胞因子TNF-α、IL-6、CXCL1、MIP-2表达,其中芪参合剂中剂量组效果最明显。结论:芪参合剂能通过调节固有免疫反应对脓毒症大鼠起到保护作用,其机制可能与降低主要脏器细菌负荷,减轻免疫细胞的过度活化,减少炎症因子的过度释放有关。

Objective:To investigate Qishen mixture in the regulation of innate immunity of drug-resistant pneumoniae Klebsiella sepsis rats.Methods:All 50 SD rats were randomly allocated to normal group,the model group,low,moderate and high dose groups of Qishen mixture,rat models with sepsis were duplicated by tracheal injection of drug-resistant pneumoniae Klebsiella,the rats were drenched with Qishen mixture or physiological saline six hours after modelling,BALF was taken out three days after the intervention,to detect total white blood cells(WBC)counting and differential WBC counting;BALF,blood,spleen and kidney were got to measure colony forming unit(CFU);ELISA method was used to detect the expressions of TNF-αand IL-6 in BALF,and CXCL1 and MIP-2 in lung tissue.Results:Total WBC counting and differential WBC counting rose notably in the BALF of the model group,bacterial load in the tissues of BALF,blood,spleen and kidney increased notably,and the expressions of TNF-α,IL-6,CXCL1 and MIP-2 in the tissue rose notably.The intervention with Qishen mixture in different dosages could reduce total WBC counting and differential WBC counting in BALF to different extents,relieve bacterial loads in the main viscera and lower the expressions of TNF-α,IL-6,CXCL1 and MIP-2 obviously,cespecially,the effects in moderate dose group of Qishen mixture the most notable.Conclusion:Qishen mixture could protect sepsis rats through regulating innate immunity,and its mechanism might be related to reducing bacterial load of the main viscera,relieving overactive immunological cells and the excessive release of inflammatory factors.