| Objective:Through the establishment of asthma rat model,explore the effect of Jianpi Bufei Huatan Formula combined with hormone on airway inflammation,cell apoptosis,oxidative stress and Keap1/Nrf2/HO-1 signal pathway in asthma rats,and further explore the mechanism of Jianpi Bufei Huatan Formula combined with hormone on asthma.Method:Sixty rats were randomly divided into 6 groups using EXCEL software,with10 rats in each group,including normal control group(ZCDZ group),asthma model group(XCMX group),dexamethasone observation group(DMGC group),low dose of Jianpi Bufei Huatan Formula combined with dexamethasone group(D-JPBF group),medium dose of Jianpi Bufei Huatan Formula combined with dexamethasone group(ZJPBF group),and high dose of Jianpi Bufei Huatan Formula combined with dexamethasone group(G-JPBF group).In addition to ZCDZ group,rats in ZCDZ group were sensitized by intraperitoneal injection of ovalbumin(OVA)on the 1st and 8th day,and rats in ZCDZ group were replaced by normal saline;From the 15 th day,except for the ZCDZ group,rats in other groups were given ovalbumin atomization to stimulate asthma attack once every other day for 8 weeks;At the same time,starting from the15 th day,once a day,the rats in the experimental groups were given the drug administration experiment.The rats in the ZCDZ group and XCMX group were replaced with normal saline,and the other groups were given the corresponding western medicine by intraperitoneal injection and traditional Chinese medicine by gavage according to the calculated dose,lasting for 8 weeks.After the experiment,the serum,BALF,lung tissue and other samples of the experimental rats were taken for subsequent detection.The lung tissue structure of the experimental rats was observed by HE staining,the apoptosis of the bronchial epithelial cells of the experimental rats was observed by TUNEL staining,and the TNF in the serum and BALF of the experimental rats was detected by ELISA-α The expression of IL-6,ROS,MDA,SOD,CAT,GSH in serum and BALF of experimental rats were detected by biochemical method,the expression of Keap1,Nrf2,HO-1 protein in lung tissue of experimental rats was detected by WB method,and the expression of Keap1,Nrf2,HO-1 m RNA in lung tissue of experimental rats was detected by RT-PCR.Result:1.General conditions of experimental rats: rats in ZCDZ group were normal in spirit,growth and development,diet and daily activities,and no obvious abnormal symptoms;XCMX group was in low spirits,growth and development slightly lagged behind that of ZCDZ group,slow response,activity and food consumption gradually decreased,and even appeared sparse dry hair,as well as nasal congestion,sneezing,slightly short irregular breathing,scratching the ears and cheeks,licking limbs,restlessness and other symptoms;The symptoms of rats in G-JPBF group,Z-JPBF,D-JPBF group and DMGC group were improved compared with those in XCMX group.There was no obvious or slight shortness of breath,restlessness was still improved,and hair gloss was also improved,but there were still significant differences compared with ZCDZ group.2.Lung tissue structure of experimental rats: the lung tissue structure of rats in the ZCDZ group was relatively normal,the trachea wall structure at all levels was complete and the epithelial structure was complete,without airway mucosal edema,the alveolar septa were normal,the structure was clear,and there was no obvious inflammatory cell infiltration.In XCMX group,the lung tissue of rats showed consolidation,the bronchial lumen was significantly narrowed,the mucosa epithelium showed edema and thickening,the alveolar lumen became smaller,the cell components in the alveolar septum increased,and a large number of inflammatory cells infiltrated around the trachea and blood vessels at all levels.Compared with XCMX group,the lung tissue structure of rats in the treatment group was improved,and the lung tissue structure of rats in the G-JPBF group was closer to that in the ZCDZ group.3.The apoptosis of bronchial epithelial cells in experimental rats: compared with ZCDZ group,the average optical density of bronchial epithelial cells in XCMX group was higher;Compared with XCMX group,the mean optical density of bronchial epithelial cells in DMGC group decreased;Compared with DMGC group,the mean optical density of bronchus of rats in G-JPBF group decreased the most.4.TNF in serum and BALF of experimental rats-α、 IL-6 expression: compared with ZCDZ group,TNF in serum and BALF of rats in other groups-α、 The content of IL-6increased;Compared with XCMX group,TNF in serum and BALF of rats in each treatment group-α、 The content of IL-6 decreased;Compared with DMGC group,TNF in serum and BALF of rats in D-JPBF group-α 、 The expression of IL-6decreased the most.5.The expression of ROS,MDA,SOD,CAT and GSH in serum and BALF of experimental rats: compared with ZCDZ group,the expression of ROS,MDA in serum and BALF of other groups increased,while the expression of SOD,CAT and GSH decreased;Compared with XCMX group,the expression of ROS and MDA in serum and BALF of rats in the treatment group decreased,and the expression of SOD,CAT and GSH increased;Compared with DMGC group,the expression of ROS and MDA in serum and BALF of rats in G-JPBF group decreased the most,and the expression of SOD,CAT and GSH increased the most.6.The expression of Keap1,Nrf2 and HO-1 protein in lung tissue of experimental rats:compared with ZCDZ group,the expression of Keap1 protein in lung tissue of experimental rats in other groups decreased,and the expression of Nrf2 and HO-1protein increased;Compared with XCMX group,the expression of Keap1 protein in lung tissue of rats in the treatment group increased,and the expression of Nrf2 and HO-1 protein decreased;Compared with DMGC group,the expression of Keap1 protein in the lung tissue of rats in G-JPBF group increased the most,and the expression of Nrf2 and HO-1 protein decreased the most.7.The expression of Keap1,Nrf2 and HO-1 m RNA in lung tissue of experimental rats:compared with ZCDZ group,the expression of Keap1 m RNA in lung tissue of experimental rats in other groups decreased,and the expression of Nrf2 and HO-1m RNA increased;Compared with XCMX group,the expression of Keap1 m RNA in lung tissue of rats in the treatment group increased,and the expression of Nrf2 and HO-1 m RNA decreased;Compared with DMGC group,the expression of Keap1 m RNA in lung tissue of rats in G-JPBF group increased the most,and the expression of Nrf2 and HO-1 m RNA decreased the most.Conclusion:1.Jianpi Bufei Huatan Decoction combined with hormone has definite therapeutic effect on asthma rats.2.Jianpi Bufei Huatan Decoction combined with hormone can significantly improve the structure of lung tissue in asthmatic rats.3.Jianpi Bufei Huatan Decoction combined with hormone can effectively reduce the apoptosis of bronchial epithelial cells in asthmatic rats.4.Jianpi Bufei Huatan Decoction combined with hormone can regulate TNF in serum and BALF of asthmatic rats-α、 The content and expression of IL-6 can further prove that the treatment of integrated traditional Chinese and western medicine has a role in reducing airway inflammation in asthmatic rats.5.Jianpi Bufei Huatan Decoction combined with hormone can reduce the expression of ROS and MDA in serum and BALF of asthmatic rats,increase the expression of SOD,CAT and GSH,and reduce the oxidative stress reaction of asthmatic rats.6.The combination of Jianpi Bufei Huatan Formula and hormone may improve the lung tissue structure of asthmatic rats,reduce the apoptosis of bronchial epithelial cells,reduce airway inflammation,and reduce oxidative stress by regulating the Keap1/Nrf2/HO-1 signal pathway. |
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