| Objective: To investigate the effect of Liuwei Buqi Formula on COPD cell senescence and PI3K/Akt/mTOR signaling pathway.This paper attempts to reveal the targets,links,and approaches of the Liuwei Buqi Formula in the treatment of COPD to obtain evidence for further optimizing the treatment of COPD.Methods: 60 rats were divided into six groups using the random table method:standard control group(NG),model group(MG),Guilong cough and allergy group(PC),and Liuwei Buqi Formula short-action group(LD),moderately dose group(MD),and long-action group(HD),each group consisted of ten rats.The remaining rats were modeled with COPD by combining LPS with cigarette smoke fumigation.On days 1 and15,the rats were anesthetized with 3% pentobarbital and smoked with cigarette smoke on the rest of the days for 5 weeks.Measurement of lung function at the end of moldinging.0.01ml/(g-d)of saline was delivered to rats in the NG and MG groups,while the rats in the PC specially prepared Guilong cough and allergy kapsul,0.62 g/kg-d,and the rats in the LD,MD,and HD groups were given Liuwei Buqi Formula suspension 0.28 g/kg-d and 0.55g/kg-d,and 1.07 g/kg-d were administered to the rats in LD,MD,and HD groups,respectively.The rats were observed daily for changes in biological characteristics.After the pharmacological intervention,lung function was assessed once more.HE staining was employed to observe the morphological alterations in the lung tissue.IL-6 and TNF-αlevels in serum and BALF were measured by ELISA,SOD levels in serum and BALF were measured by WST-1,MDA levels in serum and BALF were measured by TBA,pPI3 K,p-Akt,p-mTOR,SIRT-1,p53 and p21 mRNA levels in lung tissue were measured by Western Blot.mTOR,SIRT-1,p53 and p21 protein amounts by Western Blot,and PI3 K,Akt,mTOR,SIRT-1,p53 and p21 mRNA levels by RT-qPCR.Results:1.Rats in the NG group were full of energy,had bright skin color,normal respiration,diet,and excretion,and increased body weight.Compared with NG,the rats in the MG group were depressed and withered,with hand and hair loss,shortness of breath,open mouth breathing,sputum sounds,loss of appetite,irregular bowel movements,and weight loss.2.Lung function changes: The MG group’s FEV0.3,FEV0.3/FVC,and PEF results were substantially lower when compared to the NG group(P<0.05);the levels of FEV0.3,FEV0.3/FVC,and PEF increased to varying degrees in the PC,LD,MD,and HD groups if compared to the MG group(P<0.05);in contrast to the PC group,the LD and MD groups’ FEV0.3,FEV0.3/FVC,and PEF values rose in it(P<0.05),FEV0.3,FEV0.3/FVC,and PEF values were not altered markedly in the HD group(P>0.05);nevertheless,they did rise in the LD and MD groups as compared to the HD group(P<0.05).3.Histopathological changes of the lung: In the NG group,the lumen of the trachea and bronchi were normal,the mucosa of fine bronchi was intact and intact with no adhesion of cilia;the glandular structure was intact,the alveoli were well-shaped and normal in size,and the alveolar septum was intact without abnormality ies.Compared with the NG group,the bronchial structure of the MG group was destroyed,the lumen was narrowed,the bronchial mucosa was incomplete and exfoliated,the cilia were irregularly arranged and inverted,the glands were hypertrophic and hyperplastic,massive inflammatory infiltration had been observed in the upper and lower lung’s interstitial area and alveolar chamber,and the alveolar wall was collapsed and thinned.The PC,LD,MD,and HD groups displayed an assortment of distinctions from the MG group,which is declining in inflammatory infiltrates and improving in lung histopathological changes in the alveolar cavity and interstitium,with the most obvious improvement in the HD group.4.The serum and BALF levels’ of IL-6,TNF-α,SOD,and MDA: MDA,IL-6,and TNF-a levels were dramatically higher in the MG group compared to the NG group(P<0.05),and SOD levels were significantly minimized(P<0.05),suggesting the fact that the rats’ COPD model had been effectively constructed;compared with the MG group,decreasing in MDA,IL-6,TNF-α levels in the PC,LD,MD and HD groups(P<0.05)and the levels of SOD increased to different degrees(P<0.05),suggesting a certain therapeutic effect of drug intervention;the levels of MDA,IL-6,and TNF-in the bloodstream and the BALF of rats in the LD and MD groups increased(P<0.05)as opposed to the PC group,whereas the levels of SOD diminished(P<0.05),in the HD group,the levels of IL-6,TNF-α,and MDA diminished(P<0.05),yet the levels of SOD climbed(P<0.05);in opposition to the HD group,the levels of MDA,IL-6,and TNF-αin the LD and MD groups grew(P<0.05)and the levels of SOD activity decreased(P<0.05).5.Levels for aging-related molecular proteins in lung tissue,as well as the PI3K/Akt/mTOR pathway Particularly in comparison with the NG group,the appearance of p-PI3 K,p-Akt,p-mTOR,p21,and p53,were up substantially(P<0.05)and SIRT-1was down dramatically(P<0.05)in the lung tissues from rats in the MG group;compared with the MG group,the representation of p-PI3 K,p-Akt,p-mTOR,p21,and p53 were down sharply(P<0.05)and SIRT-1 was significantly up-regulated(P<0.05)in the lung tissues from rats in the PC,LD,M,D and HD group.Remarkably decreased levels of pPI3 K,p-Akt,p-mTOR,p21,and p53 mRNA were noted(P<0.05),and SIRT-1 was significantly up-regulated(P<0.05);the transcription of p-PI3 K,p-Akt,p-mTOR,p21,and p53 was up-regulated(P<0.05)even as SIRT-1 was weaker than expected(P<0.05)in the lung tissues of mice in the HD group compared well with PC group in the PC,LD,MD,and HD groups than with the MG group.Rats in the HD group had slightly higher compared SIRT-1 expression and down-regulated PI3 K,p-Akt,p-mTOR,p21,and p53 expression in their lung tissues(P<0.05);compared with HD group,p-PI3 K,p-Akt,pmTOR,p21,p53 expression was up-regulated in lung tissues of rats in LD and MD groups(P<0.05)and SIRT-1 expression was down-regulated(P<0.05).6.PI3K/Akt/mTOR pathway,aging-related molecule mRNA levels in lung tissue:Compared with the NG,PI3 K,Akt,mTOR,p21,p53 mRNA expression was significantly up-regulated(P<0.05)and SIRT-1 mRNA expression was significantly down-regulated(P<0.05)the MG group;compared with the MG,PI3 K,Akt,mTOR,p21,p53 mRNA expression was significantly down-regulated(P<0.05)and SIRT-1 mRNA expression was significantly up-regulated(P<0.05)in lung tissues of rats in the PC,LD,MD,and HD groups compared with the MG group;compared with the PC group,PI3 K,Akt,mTOR,p21 and p53 expression was up-regulated(P<0.05)and SIRT-1 expression was downregulated(P<0.05)in the LD and MD,and PI3 K,Akt,mTOR,p21,p53 expression was down-regulated and SIRT-1 was up-regulated in the HD(P<0.05);compared with the HD,PI3 K,Akt,mTOR,p21,p53 expression was up-regulated(P<0.05)and SIRT-1expression was down-regulated in lung tissues of rats in LD and MD groups(P<0.05).Conclusions :1.Liuwei Buqi Formula can relieve lung histopathological changes and increase lung function in COPD model rats.2.Liuwei Buqi Formula can relieve inflammation and oxidative stress in the lung tissue of COPD model rats.3.Liuwei Buqi Formula can delay the aging of lung tissue cells and increase lung function in COPD model rats.The mechanism is realized by regulating the PI3K/Akt/mTOR signaling pathway. |
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