益肺健脾方对慢性阻塞性肺疾病大鼠血清和支气管肺泡灌洗液中炎症因子水平的影响

Effect of Yifei Jianpi Prescription on the Levels of Inflammatory Factors in Serum and Bronchoalveolar Lavage Fluid of Rats with Chronic Obstructive Pulmonary Disease

目的观察益肺健脾方对慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)模型大鼠血清和支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中炎症因子水平的影响。方法将SD大鼠随机分为空白组,模型组,益肺健脾方低、中、高剂量组,地塞米松组和氨溴索组;除空白组外,其余6组大鼠均采用熏烟、气管内滴注脂多糖及番泻叶灌胃多因素联合制备肺脾两虚型COPD大鼠模型,共49 d。模型复制完成后,给予相应药物灌胃6周:空白组大鼠每日予生理盐水10 mL/kg灌胃;益肺健脾方低、中、高剂量组大鼠每日分别予益肺健脾方药液0.8、1.6、3.2 g/kg灌胃;地塞米松组大鼠每日予地塞米松混悬液0.05252mg/kg灌胃;氨溴索组大鼠每日予氨溴索混悬液1.4 mg/kg灌胃。采用苏木精—伊红染色法观察大鼠右肺下叶及支气管病理变化;阿尔辛蓝—过碘酸雪夫(Alcian blue-periodic acid Schiff,AB-PAS)染色法观察大鼠支气管杯状细胞增生情况;ELISA法检测大鼠血清和BALF中白细胞介素(interleukin,IL)-8、IL-10、IL-17、肿瘤坏死因子(tumor necrosis factor,TNF)-α、单核细胞趋化蛋白-1(monocyte chemoattractant protein-1,MCP-1)、瘦素(leptin)、C反应蛋白(C-reactive protein,CRP)、基质金属蛋白酶-9(matrix metallo proteinase-9,MMP-9)水平。结果苏木精—伊红染色结果显示,益肺健脾方高剂量和地塞米松能显著改善COPD大鼠气道重塑、炎症细胞浸润;AB-PAS染色显示,各用药组大鼠支气管杯状细胞增生明显减少。益肺健脾方高剂量能显著降低COPD大鼠血清和BALF中IL-8、IL-17、MCP-1、leptin、TNF-α、CRP、MMP-9水平(P<0.05),升高IL-10水平(P<0.05)。结论益肺健脾方能有效减轻大鼠气道炎症,改善气道重塑,其机制可能与调控IL-8、IL-10、IL-17、MCP-1、leptin、TNF-α、CRP、MMP-9水平有关。

Objective To investigate the effect of Yifei Jianpi prescription on the levels of inflammatory factors in serum and bronchoalveolar lavage fluid(BALF)of rats with chronic obstructive pulmonary disease(COPD).Methods Sprague-Dawley rats were randomly divided into blankgroup,model group,dexamethasone group,ambroxol group,and low-,middle-,and high-dose Yifei Jianpi prescription groups.All rats except those in the control group were exposed to tobacco smoke and were given intratracheal instillation of lipopolysaccharide and intragastric administration of senna for 49 days to establish a rat model of COPD with deficiency of both lung and spleen.After modeling,the drugs were given by gavage once a day for 6 weeks:the rats in the blank group were given normal saline 10 mL/kg;the rats in the low-,middle-,and high-dose Yifei Jianpi prescription groups were given Yifei Jianpi prescription at a dose of 0.8,1.6,and 3.2 g/kg,respectively;the rats in the dexamethasone group were given dexamethasone suspension 0.0525 mg/kg;the rats in the ambroxol group were given ambroxol suspension 1.4 mg/kg.Hematoxylin and eosin(HE)staining was used to observe the pathological changes of the right inferior lobe and the bronchus,and Alcian blue-periodic acid Schiff(AB-PAS)staining was used to observe the proliferation of goblet cells in the bronchus of rats;ELISA was used to measure the levels of interleukin-8(IL-8),interleukin-10(IL-10),interleukin-17(IL-17),tumor necrosis factor-α(TNF-α),monocyte chemotactic protein-1(MCP-1),leptin,C-reactive protein(CRP),and matrix metallopeptidase-9(MMP-9)in serum and BALF.Results HE staining showed that high-dose Yifei Jianpi prescription and dexamethasone significantly improved airway remodeling and inflammatory cell infiltration in rats with COPD,and AB-PAS staining showed that there was a significant reduction in the proliferation of goblet cells in the bronchus of rats in each administration group.High-dose Yifei Jianpi prescription significantly reduced the levels of IL-8,IL-17,MCP-1,leptin,TNF-α,CRP,and MMP-9(P<0.05)and increased the level of IL-10(P<0.05)in serum and BALF of COPD rats.Conclusion Yifei Jianpi prescription can effectively alleviate airway inflammation and improve airway remodeling in rats,possibly by regulating the levels of IL-8,IL-10,IL-17,MCP-1,leptin,TNF-α,CRP,and MMP-9.