Effects Of Chai Pu Tang-containing Serum On The Expression Of MAL, CADM1 And TSLP In Airway Epithelial Cells Of Mice Induced By Asthma Seru

Objective:To investigate the effect of Chaipu decoction containing serum on theexpression of CADM1,MAL,TSLP,YKL-40 in mouse airway epithelial cells induced by liver depression asthma serum.Methods: The mouse model of liver depression and asthma was established to obtain liver depression asthma serum and Chai Pu decoction medicated serum.The medicated serum and liver-stagnation asthma serum were cocultured with mouse airway epithelial cells.To detect the effect of Chaipu decoction containing serum on the expression of CADM1,MAL,TSLP,YKL-40 in mouse airway epithelial cells of mice.The first stage: 24 clean Balb/c mice weighing 20±2g were adaptively fed for 1 week,and they were randomly divided into three groups: normal group,liver depression asthma group,and Chaipu decoction group.The model of liver depression and Chaipu decoction group was established by chronic mild and unpredictable stress(CUMS)method from 1 to 28 days.On the 29 th and 42 nd day,the mice were sensitized by abdominal injection of ovalbumin(OVA)and aluminum hydroxide.On the 43 rd day,each mouse in each group was separately placed in a semi-sealed atomization box.The normal group was atomized with normal saline.The mice in the liver depression asthma group and Chaipu decoctiongroup were stimulated with 1% OVA atomization once a day for half a hour for 7 days.The mouse model of liver depression asthma was established.The mouse model of liver depression asthma was established.Chaipu decoction 0.3ml was injected into the stomach half an hour before each atomization,and the normal group and the liver depression asthma group were given the same amount of normal saline.The second stage: 12 hour after the last administration,the mice in each group were anesthetized and blood was collected from the orbit,and the serum was separated.The obtained taken were normal serum,liver depression asthma serum and Chai Pu decoction medicated serum.After blood was taken from the orbit,the lung tissue of the mouse was taken to make a slice for HE staining detection.The third stage: The purchased normal mouse bronchial epithelial cells were divided into normal group,liver depression asthma group,Chaipu decoction group.The normal group was treated with 2 ml of normal group serum;the liver depression asthma group was treated with liver depression asthma serum 2ml treatment;the Chaipu decoction group was treated with 2 ml of liver depression asthma serum +2 ml of Chaipu decoction containing serum.The serum of each experimental group was configured at a concentration of 20%,and the same amount of medium was added in each group.After 24 hours of culture,the cell culture medium and cells were collected.The fourth stage:The morphological changes of cells were observed by HE staining microscope and image analysis system.The concentration of TSLP and YKL-40 in the culture supernatant of airway epithelial cells was detected by ELISA method.The expression of MAL,CADM1,TSLP and YKL-40 protein was detected by Western Blot,and the expression of MAL,CADM1,TSLP and YKL-40 mRNA was detected by RT-q PCR.Results: 1.HE :the airway of the mice in the normal group was not abnormal,the lumen structure was regular,and the airway wall was not thickened.The airway structure of the mice in the liver depression asthma group was irregular,the lumen was narrow,and the tube wall was thickened,there were a large number of inflammatory cell infiltration in the lumen,goblet cell degeneration and proliferation,a large amount of mucus secretion,epithelial cell arrangement disorder,extracellular matrix deposition,airway smooth muscle hypertrophy and hyperplasia.The bronchial wall in the Chaipu decoction group was smoother,and pathological changes in Chaipu decoction group were significantly less than those in liver depression asthma group.ELISA: The concentrations of TSLP and YKL-40 in the airway epithelial cell culture supernatant of mice in the liver depression asthma group were significantly higher than those in the normal group(P<0.05),and the airway of the mice was treated with Chaipu decoction containing serum.The concentration of TSLP and YKL-40 in the epithelial cell culture supernatant was reduced significantly(P<0.05).3.WB: The expression of CADM1 protein in the liver depression asthma group was lower than that in the normal group(P<0.05),between chaipu Decoction group and liver stagnation asthma group.The expression of MAL protein in the liver depression asthma group was lower than that in the normal group(P<0.05),and the expression of MAL protein in chaipu Decoction group was higher than the liver depression asthma group(P<0.05).The protein expressions of TSLP and YKL-40 in the liver-stagnation asthma group were higher than those in the normal group.The protein expression of TSLP and YKL-40 in chaipu Decoction group were lower than that in the liver depression asthma group.4.RT-qPCR: The expression of CADM1 mRNA in the liver depression asthma group was lower than that in the normal group(P<0.05),between chaipu Decoction group and liver stagnation asthma group.The expression of MAL mRNA in the liver depression asthma group was lower than that in the normal group(P<0.05),and the MAL mRNA expression in chaipu Decoction group was higher than that in the liver stagnation asthma group(P<0.05).The expression of TSLP and YKL-40 mRNA in the liver depression asthma group was higher than that in the normal group.The expression of TSLP and YKL-40 mRNA in chaipu Decoction group were lower than that in the liver depression asthma group.Conclusion: 1.Liver depression asthma serum can reduce the expression of CADM1 and MAL in airway epithelial cells to decrease,while the expression of TSLP and YKL-40 to increase.2.Chaipu decoction containing serum can reduce the expression of TSLP and YKL40 and raise the expression of MAL in airway epithelial cells cultured with liver depression asthma serum.