Pingchuanning Regulates IRE-1α/XBP-1s/NF-κB P65 In Lung Tissue Of Rats With Cold Asthma Study On The Effect Mechanism Of On Airway Inflammation

Objective:the purpose of this experiment is to explore the effects of Pingchuanning on airway morphology,inflammatory factors IL-5,IL-17 in BALF,and the effect of the expression level of Hgsnat、Pdgfrb、Scara3、IRE-1α、XBP-1s、NF-κB p65 in lung tissue,and then study the possible mechanism of Pingchuanning on reducing the airway inflammation of bronchial asthma,so as to expand the target of Pingchuanning in treating bronchial asthma,so that this prescription can be better used in clinical practice.Methods:A total of 105 SPF healthy male SD rats weighing 150-180g and aged 6-8weeks were randomly divided into 7 groups.They were divided into normal group(NC),model group(MC),low dose group(PCN-L),medium dose group(PCN-M),high dose group(PCN-H),Guilong Kechuanning group(GK)and dexamethasone group(DEX),with 15 animals in each group.The model was built according to the design requirements and methods of cold croup model.First,the rats were fed regularly for a week.On the first and eighth days after a week of routine feeding,the rats in MC group,Psn-L group,PSN-M group,PSN-H group,GK group and DEX group were intraperitoneally injected 1ml 10%ovalin suspension(made of ovalin,dry aluminum hydroxide powder and normal saline)to sensitized;From the 15th to the 21st day,the six groups of rats except the normal group were stimulated by ultrasonic atomization with 1%ooprotein suspension at 30 minutes/day,and the rats were placed in a cold box for the same time and the same number of cold stimulation,so as to build a cold asthma model.In NC group,the same volume of normal saline was used for intraperitoneal injection and atomization.21 days after modeling,the rats in normal group and model group were given the same amount of normal saline intragastric administration every day.The rats in Pingchanning low-dose,medium-dose and high-dose groups,dexamethasone group and Guilong Kechuanning group were converted into the equivalent dose of rats according to the normal adult dose,and the rats were given intragastric administration according to the dose conversion ratio.The proportion of the rats in low-dose,medium-dose and high-dose Pingchanning groups was converted to1:2:4.The rats were given intragastric treatment,once a day,and after continuous intragastric administration for 28 days,the rats were anesthetized by intraperitoneal injection of 20mg/100g sodium barbiturate with a concentration of 2%.The rats were sacrificed after fixed sampling according to the test requirements.During the modeling process,attention should be paid to recording the changes of body weight,mental state and death of rats in each group.HE staining was used to observe the pathological changes of smooth muscle thickness and inflammatory cell infiltration degree in lung tissue.The expressions of IL-5 and IL-17 in the supernatant of BALF were detected by ELISA.The expression levels of HgsnatmRNA,PdgfrbmRNA,Scara3mRNA,IRE-1αmRNA,XBP-1smRNA and NF-κB p65mRNA in lung tissues were detected by RT-PCR.The expressions of IRE-1α,XBP-1s and NF-κB p65 in lung tissues of rats were detected by Western blot.Results:1.HE staining:In the normal group(NC),the lung tissue structure of rats was clear,the bronchial cavity structure was complete,the alveolar structure was complete,the lung bullae showed normal shape,no obvious abnormal inflammatory cell infiltration was observed,and there was no mucous substance in the cavity.Compared with the normal group(NC),the bronchial walls of rats in the model group(MC)were significantly thickened,the mucosa folds increased,the lumen was narrow and a large amount of mucus was visible,the perialveolar interstitium was significantly exudative edema,the alveolar collapse was serious,and a large number of inflammatory cells were infiltrated in the vascular and peribronchial area.Compared with model group(MC),Pingchanning high-dose,medium-dose and low-dose groups(PCN-H,M,L),Guilong Kechuanning group(GK)and dexamethasone group(DEX)were improved to varying degrees.2.ELISA detection:Compared with NC group,the expression levels of IL-5 and IL-17in bronchoalveolar lavage fluid(BALF)of rats in MC group were significantly increased(P<0.01);Compared with MC group,the expression levels of IL-5 and IL-17in BALF of rats in each drug intervention group showed a significantly decreased trend(P<0.01).3.RT-qPCR detection:Compared with NC group,the expression levels of IRE-1αmRNA,XBP-1smRNA and NF-κB p65 mrna in MC group were significantly increased(P<0.01).The expression levels of Hgsnat mRNA,Pdgfrb mRNA and Scara3mRNA were decreased(P<0.01).Compared with MC group,the expression levels of IRE-1αmRNA,XBP-1s mRNA and NF-κB p65mRNA in lung tissue of rats in drug intervention groups were significantly decreased(P<0.01).The expression levels of Hgsnat mRNA,Pdgfrb mRNA and Scara3mRNA showed an increasing trend(P<0.01,P<0.05).4.Western blot assay:Compared with NC group,the expression levels of IRE-1α,XBP-1s and NF-κB p65 in lung tissue of MC group were significantly increased(P<0.01).Compared with MC group,the expression levels of IRE-1α,XBP-1s and NF-κB p65 in lung tissue of rats in drug intervention groups were significantly decreased(P<0.01).Conclusion:Pingchuanning can regulate The expression of IRE-1α,XBP-1s,NF-κB p65 in lung tissue of rats with cold asthma,can also regulate IRE1 dependent decay(RIDD)response,thereby inhibiting airway inflammation,relieving airway remodeling,and improving asthma symptoms in cold asthmatic rats.