Mechanism Research On The Intervention Remodeling Airway Of PI3K-Akt/PKB By Applying Pingchuanning To Regulating P85?mTOR?ASK1 Which Existing In The Lung Tissues Of Asthma Rats

Objective: In this study,we investigated the effect of Pingchuanning on PI3K-Akt/PKB signal transduction pathway related proteins in lung tissue of cold asthmatic rats to explore the mechanism of airway remodeling in asthma.The expression of P85,m TOR,ASK1,Caspase 8 and TSC1 in lung tissue of cold asthma rats was detected by immunohistochemistry and fluorescence quantitative PCR.The effect of Pingchuanning on the expression of P85,m TOR,ASK1,Caspase 8 and TSC1 was also studied to explore the mechanism of Pingchuanning on delaying and improving airway remodeling in cold asthma rats.Method: 1.105 healthy male SD rats aged 6-8 weeks and weighing 180-230 g were selected and purchased from the Experimental Animal Center of Anhui Medical University.They were randomly divided into seven groups: normal group,model group,Pingchuanning high-dose group,Pingchuanning medium-dose group,Pingchuanning low-dose group,Guilong Kechuanning group and dexamethasone group.2.Rats were sensitized by intraperitoneal injection of 1 ml of suspension(composed of normal saline and 10% ovalbumin)on the 1st and 8th days after a week of adaptive feeding.In the normal group,1 ml of normal saline was used to replace the suspension for intraperitoneal injection.3.From the 15 th day,the rats of the other six groups except the normal group were subjected to continuous atomization allergy for one week to prepare the cold asthma model.The specific operation was as follows: the rats were subjected to ultrasonic atomization with 1% ovalbumin suspension,and the rats were subjected to cold stimulation for 30 min every day.After 22 days,rats were given intragastric administration.The dosage was converted according to the body surface area of rats and human,and maintained for 4 weeks.Normal group and model group were given the same method to get normal saline for intragastric administration.4.After 28 days of continuous intragastric administration,ovalbumin was stimulated for the last time in rats.Within 2 hours after stimulation,the rats were dissected and their lungs were taken out.The left lungs of rats were fixed in formalin and the right lungs were frozen in a refrigerator at-80 C.HE staining was used to observe the pathological changes of lung tissue in experimental rats,immunohistochemistry was used to detect the expression of P85 and m TOR in lung tissue,and RT-q PCR was used to detect the expression of ASK1,TSC1 and Caspase 8.Results: After HE staining,compared with the normal group,the bronchial structure of the model group was changed,and a variety of inflammatory cell infiltration,mucus embolism and exfoliated bronchial epithelial cells were observed.The smooth muscle of the trachea was obviously proliferated,mucosal folds were increased,and the lumen was narrowed.The airway remodeling of the remaining five drug groups was improved to varying degrees compared with the model group.RT-q PCR detection: Compared with the normal group,the expression of ASK1 and Caspase 8 in the other six groups increased(P<0.01),with the most significant increase in the model group(P<0.01);compared with the model group,the expression of ASK1 and Caspase 8 in each drug group decreased(P<0.01),and the expression of ASK1 and Caspase 8 in the high dose group of Pingchuanning was significantly lower than that in the dexamethasone group and Guilong Kechuanning group(P<0.01)?Compared with the normal group,the expression of TSC1 in the other six groups decreased(P<0.01),with a significant decrease in the model group(P<0.01);compared with the model group,the expression of TSC1 in each drug group increased(P<0.01);the expression of TSC1 in Pingchuanning high-dose group was higher than that in dexamethasone group and Guilong Kechuanning group(P<0.01),and the expression of TSC1 in these two groups was higher than that in Guilong Kechuanning group(P<0.01).Tiao was higher than Pingchuanning medium and low dose group(P<0.01).Immunohistochemical assay: Compared with the normal group,the expression of p85 in the other six groups decreased in varying degrees(P<0.01),especially in the model group(P<0.01).Compared with the model group,the expression of p85 increased in the other five drug treatment groups(P<0.01),and the up-regulation of p85 expression in the high and middle dose groups of Pingchuanning was higher than that in the dexamethasone group and Guilong Kechuanning group(P<0.01),while the up-regulation of p85 expression in the two groups was higher than that in the low dose group of Pingchuanning(P<0.01).Compared with the normal group,the expression of m TOR increased in the other six groups(P<0.01),especially in the model group(P<0.01).Compared with the model group,the expression of m TOR in the other five drug groups was down-regulated in varying degrees(P<0.01).The down-regulated expression of m TOR in the high and middle dose groups of Pingchuanning was better than that in the dexamethasone group and Guilong Kechuanning group(P<0.01).Conclusion: Pingchuanning can reduce airway inflammation and reduce airway hyperresponsiveness by regulating P85,m TOR,ASK1,TSC1 and Caspase8 in PI3K-Akt/PKB signaling pathway in rats with cold asthma.And can reduce the effect of airway remodeling to achieve asthma treatment.