Study On The Mechanism By Yanghe Pingchuan Granules Improve Airway Remodeling Of Bronchialasthma Based On PI3K-Akt/PKB Signaling Pathway

Yanghe Pingchuan Granulesis(YPG)is the hospital preparation of the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine,and it is created on the basis of “Yanghe decoction”,“Pingchuan decoction”,the concept of“Invigorating the kidney and resolving phlegm” and years of clinical practice.The formula contains nine Chinese herbal medicines that can relieve cough and phlegm,and treat kidney-Yang deficiency from lung and kidney.Objective YPG is mainly used for the treatment of bronchial asthma(bronchial asthma,BA),and the curative effect is accurate,but the specific mechanism of the effect is not clear.This research is based on the PI3K/PKB signal pathway,to explore the potential molecular mechanism of Yanghe Pingchuan Granule in the treatment of BA.Methods1.Regulatory Effect of YPG on PI3K/PKB Signal Pathway in BA Rats with“Kidney-Yang Deficiency”BA rat model was induced by intraperitoneal injection and nebulization of ovalbumin(OVA),based on the mode of sensitization re-excitation.Ten normal rats were selected as the normal group,and the other BA rats were divided into the model group,the YPG administration group(14.76,7.38,3.69 g/kg)and Guilong Kechuanning group(1.0g/kg),10 rats in each group.Two weeks after the administration,histological ananlysis of bronchi tissue was by hematoxylin-eosin staining(HE),Realtime-PCR method was undertaken to determine the expression of phosphatidylinositide 3-kinases(PI3K),protein kinase B(PKB),adrenergic receptor(AR),and Proliferating Cell Nuclear Antigen(PCNA).Western blot(WB)method were undertaken to determine the expression of p-PI3 K,p-PKB,p-AR,p-PCNA protein.ELISA was used to detect the expression of phosphatidylinositol(3,4)bisphosphate(PI(3,4)P2),phosphatidylinosito(3,4,5)trisphosphate(PI(3,4,5)P3),epinephrine(EPI)and interleukin-type inflammatory cytokines.2.Regulation of serum of YPG on PI3K/PKB signaling pathway of ASMCs in OVA induced BA rats.BA rat model was induced by intraperitoneal injection and nebulization of ovalbumin(OVA),based on the mode of sensitization re-excitation.The primary cells of bronchial airway smooth muscle(ASMCs)in BA rats were taken and cultured for third generation cells,MTT method for screening the optimal concentration and the best action time of serum of YPG,ASMCs were divided into normal group,model group,serum of YPG group,PI3K/PKB pathway blocker group and serum of YPG+blocker combination group.After two weeks of intervention,Realtime-PCR method was undertaken to determine the expression of phosphatidylinositide 3-kinases(PI3K),protein kinase B(PKB)and Proliferating Cell Nuclear Antigen(PCNA).Western blot(WB)method were undertaken to determine the expression of p-PI3 K,p-PKB protein.ELisa was used to detect the expression of phosphatidylinositol(3,4)bisphosphate(PI(3,4)P2),phosphatidylinositol(3,4,5)trisphosphate(PI(3,4,5)P3)and interleukin-type inflammatory cytokines.Result1.Regulatory Effect of YPG on PI3K/PKB Signal Pathway in BA Rats with“Kidney-Yang Deficiency”Compared with the model group,the degree of lung lesion in the rats of each treatment group was improved,and it was dose-dependent.And YPG can significantly reduce the expression of PI3 K,PKB,PIP2,PIP3,PCNA and inflammatory mediators in BA rats lungs in a dose-dependent manner,and up-regulate the levels of EPI and AR;2.Regulation of serum of YPG on PI3K/PKB signaling pathway of ASMCs in OVA induced BA rats.The optimal concentration and the best action time of serum of YPG was screened by MTT method,and the optimal concentration and the best action time was 50% and 48 h respectively.The serum of YPG group,the pathway blocker group and the combination group could obviously decrease the levels of PI3 K,PKB,PIP2,PIP3 and PCNA inASMCs,and inhibit the PI3K/PKB signaling pathway,inhibits abnormal cell proliferation.ConclusionYPG has a definite therapeutic effect on kidney-yang-deficiency bronchial asthmatic rats.Inhibition of PI3K/PKB signaling pathway conduction and treatment of kidney yang deficiency may be the potential mechanism of YPG in the treatment of BA.