Effects Of Qingfei Huatan Decoction On IL-8,IL-17 And IL-10 In COPD Model Rats

Objective:To replicate the rat model of chronic obstructive pulmonary disease(COPD)by endotracheal injection of lipopolysaccharide(LPS)combined with fumigation.The levels of IL-8,IL-17 and IL-10 in serum,bronchoalveolar lavage fluid(BALF)and MUC5 AC m RNA expression in lung tissues of COPD model rats were detected by ELISA and RT-PCR.To observe the changes of IL-8,IL-17 and IL-10 contents in serum and BALF and MUC5 AC m RNA expression level in lung tissue of the COPD model rats after Qingfei Huatan Decoction intervention,and to explore the effect of Qingfei Huatan Decoction from the aspects of airway inflammation and high secretion of airway mucus,so as to provide experimental basis for the clinical treatment of COPD patients with Traditional Chinese Medicine.1.Prepare COPD rat modelSixty SPF Wistar male rats aged 7 weeks and weighing 200±20g were randomly divided into 6 groups: group A was normal group,group B was model group,group C was Qingfei Huatan decoction low-dose group,group D was Qingfei Huatan decoction medium-dose group,group E was Qingfei Huatan decoction high-dose group,and group F was roxithromycin group,with 10 rats per group.Before the experiment,6 groups of rats were placed in the experimental environment for adaptive feeding for 1 week,free water and food intake.After 1 week of adaptive feeding,group A was fed as usual without intervention,and the other groups were modeled by endotracheal injection of LPS combined with smoking compound modeling method.LPS was dropped into the trachea of rats on the 1st and 14 th day of the experiment,and the rats were given cigarette smoking in self-made fumigation box on the2 nd to 13 th and 15 th to 28 th days to establish COPD rat model.On day 29,one rat was sacrificed in groups B,C,D,E and F for pathological examination to confirm whether the model was successful.2.Drug intervention of roxithromycin and Qingfei Huatan DecoctionGroup A and B were given 10ml/(kg·d)normal saline intragastric administration,group C,D and E were given 5.69g/(kg·d),11.37g/(kg·d)and 22.74g/(kg·d)Qingfei Huatan decoction,respectively,and group F was given 30.45mg/(kg·d)roxithromycin for 14 consecutive days.3.Specimen collection and testingAfter 14 days of administration,the same method was adopted to collect the samples of rats in each group: after anesthesia,blood samples were collected from the abdominal aorta,BALF was collected from the left lung and lung tissue was collected from the right lung.After the specimens were collected,the pathological changes of the lung tissues of rats were observed by light microscope.The contents of IL-8,IL-17 and IL-10 in the serum of the left lung BALF and blood centrifugation were determined by ELISA,and the m RNA expression level of MUC5 AC in the right lung tissues was detected by RT-PCR.Results:1.Preparation of rat model: According to the general manifestations of cough,phlegm,nasal and oral secretions,hair,activity status and other conditions of the rats,the symptoms were consistent with COPD.The right lung tissue was observed by light microscope,except group A,the pathological changes were consistent with COPD.2.Pathological scores of lung tissue inflammation:(1)Compared with group A,the inflammatory pathological reaction of lung tissue in group B was significantly higher,P<0.05.(2)Compared with group B,the inflammatory pathological reaction of lung tissue in group C,D,E and F was reduced,P<0.05.(3)Compared with group C,the pathological changes of lung inflammation in group D were not obvious,P>0.05;The pathological changes of lung inflammation in Group E were significantly reduced,P<0.05.(4)Compared with group D,the pathological changes of lung inflammation in Group E were significantly reduced,P<0.05.(5)Compared with group E,the pathological changes of lung inflammation in group F were not significant,P>0.05.3.Results of drug intervention:(1)compared with group A,the contents of IL-8 and IL-17 in serum and BALF and the expression level of MUC5 AC m RNA in lung tissue in group B were significantly increased,while the contents of IL-10 in serum and BALF were significantly decreased,P<0.05.(2)Compared with group B,the contents of IL-8 and IL-17 in serum and BALF and the expression level of MUC5 AC m RNA in lung tissue were decreased in groups C,D,E and F,while the contents of IL-10 in serum and BALF were increased,P<0.05.(3)Compared with group C,the contents of IL-8 and IL-17 in serum and BALF and the expression of MUC5 AC m RNA in lung tissue in groups D and E decreased,while the contents of IL-10 in serum and BALF increased,P<0.05.(4)Compared with group D,the contents of IL-8 and IL-17 in serum and BALF and the expression of MUC5 AC m RNA in lung tissue in group E were decreased,while the contents of IL-10 in serum and BALF were increased,P<0.05.(5)Compared with group E,the contents of IL-8,IL-17 and IL-10 in serum and BALF and the m RNA expression of MUC5 AC in lung tissue in group F were not significantly changed,P>0.05.(6)Compared with group A,the m RNA expression levels of MUC5 AC in serum,BALF,IL-8,IL-17 and IL-10 and lung tissues of rats in group E and F were significantly different(P<0.05).Conclusion:1.The COPD rat model can be successfully reproduced by intratracheal injection of LPS combined with fumigation.2.Qingfei Huatan Decoction can reduce the content of IL-8 and IL-17 inflammatory factors in serum and BALF of COPD model rats,increase the content of IL-10 anti-inflammatory medium in serum and BALF,and down regulate the expression level of MUC5 AC m RNA in lung tissue,so as to reduce airway inflammatory response and airway mucus hypersecretion,improve the pathological changes of lung tissue of COPD model rats and delay the progress of COPD.