抗复感颗粒对肺炎幼龄大鼠的干预作用及机制研究

Intervention Effect and Mechanism of Kangfugan Granule on Immature Rats with Pneumonia

目的探讨抗复感颗粒对肺炎幼龄大鼠的干预作用及机制。方法将50只大鼠随机分为正常组、模型组、头孢曲松组、抗复感颗粒组、抗复感颗粒加头孢曲松联合用药(简称联合用药)组,每组10只。运用肺炎链球菌经气管滴注构建制备幼龄大鼠肺炎模型。抗复感颗粒组给予20 g/(kg·d)抗复感颗粒灌胃及0.05 mL生理盐水肌肉注射;联合用药组给予抗复感颗粒20 g/(kg·d)灌胃及肌肉注射头孢曲松100 mg/(kg·d);头孢曲松组给予0.6 mL/d生理盐水灌胃及肌肉注射头孢曲松100 mg/(kg·d);正常组、模型组均给予等量生理盐水灌胃及肌肉注射。连续给药5天后,观察大鼠一般情况;瑞氏-姬姆萨染色对肺泡灌洗液(BALF)中细胞的总数及分类细胞数计数;HE染色观察肺及大肠组织炎症细胞浸润;q RT-PCR法检测大鼠肺组织中肿瘤坏死因子(TNF-α)、IL-6 mRNA表达水平;系列生化法检测大鼠大便肠球菌、双歧杆菌、大肠杆菌、乳酸杆菌的变化。结果与模型组比较,各药物干预组肺组织炎症细胞浸润明显改善;抗复感颗粒组肠道轻度炎症细胞浸润,联合用药组肠道无明显异常。与正常组比较,模型组大鼠体重减轻(P<0.05),白细胞总数升高,其中中性粒细胞和淋巴细胞所占比例均升高(P<0.01),巨噬细胞比例降低(P<0.01);肺组织TNF-α、IL-6 mRNA水平升高(P<0.01);肠道中肠球菌、大肠杆菌数增高,乳酸杆菌降低(P<0.01)。与模型组比较,各药物干预组白细胞总数、中性粒细胞及淋巴细胞比例均降低,巨噬细胞比例增高(P<0.01),TNF-α、IL-6 mRNA水平降低(P<0.01);抗复感颗粒组、联合用药组大鼠体重增加,肠道中肠球菌、大肠杆菌数降低,乳酸杆菌增多(P<0.05,P<0.01)。联合用药组较头孢曲松组及抗复感颗粒组白细胞总数下降(P<0.01),联合用药组较抗复感颗粒组乳酸杆菌减少(P<0.01)。结论抗复感颗粒可改善肺炎幼龄大鼠肺组织炎症,降低肺组织炎性细胞因子IL-6、TNF-αm RNA水平,且不引起肠道微生态的紊乱。

Objective To investigate the effect and mechanism of Kangfugan granules(KFG)on immature rats with pneumonia. Methods Totally 50 immature rats were randomly divided into the normal group, model group, ceftriaxone(CRO)group, KFG group and KFG+CRO group,10 in each group. The pneumonia model of immature rats were established by tracheal infusion of Streptococcus pneumoniae. Rats in KFG group were given 20 g · kg-1· d-1 KFG by gastrogavage and 0.05 mL normal saline(NS) by intramuscular injection;the combination group were given 20 g · kg-1· d-1 KFG by gastrogavage and 100 mg/kg-1· d-1 ceftriaxone by intramuscular injection;the ceftriaxone group were given 0.6 mL/d NS by gastrogavage and 100 mg/kg-1· d-1 ceftriaxone by intramuscular injection;the normal group and model group were given the equal amount of NS by gastrogavage and intramuscular injection. After 5 days of continuous administration, the general conditions of rats were observed. The total number of bronchoalveolar lavage fluid(BALF)cel s was counted and differential cells counts were quantified. The infiltration of inflammatory cell in lung and intestine tissues were observed by HE staining. The mRNA expression levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in lung tissues were detected by quantitative real-time PCR. The changes of Enterococcus, Bifidobacterium, Escherichia coli and Lactobacil us in feces were detected by serial biochemical methods.Results Compared with model group, the inflammatory cell infiltration in lung tissue was significantly improved in each intervention group. A small amount of inflammatory cell infiltration was observed in the intestinal tract in KFG group, while no obvious changes of that in lung tissue in KFG+CRO group. Compared with normal group, the weight of rats decreased(P<0.05),the total number of leukocytes, the proportion of neutrophils and lymphocytes increased, the proportion of macrophages decreased in the model group(P<0.01), the mRNA levels of TNF-α and IL-6 increased(P<0.01), the number of Enterococcus and Escherichia coli in intestinal increased, the number of Lactobacillus decreased(all P<0.01). Compared with the model group, the total number of leukocytes, the proportion of neutrophils and lymphocytes decreased, the proportion of macrophages increased(all P<0.01), the mRNA levels of TNF-αand IL-6 decreased(P<0.01)in each intervention group. The weight of rats increased,the number of Enterococcus and Escherichia coli decreased, the number of Lactobacil us increased in KFG group and KFG+CRO group(P<0.05,P<0.01). Compared with KFG+CRO group, the total number of leukocytes decreased in CRO group and KFG group(P<0.01), the number of Lactobacillus decreased in KFG group(P<0.01). Conclusion KFG could improve lung inflammation in immature rats with pneumonia, reduce the mRNA levels of IL-6 and TNF-α in lung tissue, without causing the disorder of intestinal microecology.