Research Of The Efficacy And Mechanism On Traditional Chinese Medicine Renshen-pingfei Powder

Objective:To sort out the theory source,compatibility and application characteristics of Renshen-pingfei powder(RSPF)based on the documents from both ancient and modern times.The pulmonary fibrosis rats model was duplicated by bleomycin(BLM),and transforming growth factor ?1(TGF-?1)was used to stimulate human embryonic lung fibroblast(HELF)cells.To observe the effect and mechanism of RPSF on pulmonary fibrosis rats model in vivo,as well as the regulation on epithelial-mesenchymal transition(EMT)and autophagy related protein expressions in vitro.To explore the potential mechanism of RPSF on IPF,proving scientific basis for the application of RPSF on clinic.Methods:1.Related treatises about RSPF in the literature of traditional Chinese medicine in the past dynasties were collected to sort out the theory source of RSPF.The compatibility and application characteristics of RSPF in the ZhengYinMaiZhi were discussed.The significance and value of RPSF in treating relevant syndromes of modern disease pulmonary fibrosis were analyzed by sorting out the relevant literature in the open databases of CNKI,VIP,Wanfang,etc.2.The pulmonary fibrosis model in rats was duplicated by BLM,then interfered with psilocybin and different doses of RSPF.The measurement indicators were:rats weight and death records,rats lung function parameters,lung coefficient,hydroxyproline(HYP)content in lung tissue,pulmonary histopathology changes by HE and Masson staining,mRNA and proteins expressions o EMT and autophagy related by qPCR and WB respectively.3.HELF cells were cultured in vitro,a normal control group,a lOng/mL TGF-?1 treatment group,a TGF-?1+10?g/mL RPSF group,a TGF-?1+20?g/mL RPSF group,a TGF-?1+30?g/mL RPSF group were established.MTT method was used to detect HELF proliferation.qPCR and WB methods were used to detect mRNA and protein expressions of a-SMA,E-cadherin,LC3B,Beclin-1.Results:1.RSPF was first recorded in ZhengYinMaiZhi,mainly dealing with Fei Bi due to the damage of lung Qi,and Qi deficiency.According to the review of modern literature,pulmonary fibrosis can be classified into the category of Fei Bi.RSPF is used to treat pulmonary fibrosis due to the injury of Qi and Yin and the depression of lung-heat.2.Effects of RSPF on pulmonary fibrosis rats model induced by BLM:?survival rate:the survival rate was 57%in the model group,82%in the RSPF middle and RSPF high dose group,73%in the PSPF low dose group and the positivegroup.?lung function:Compared with the model group,the value of Tidal volume(TV)was significantly increased in the RSPF middle dose group(P<0.05)and the positive group(P<0.01);the value of Ventilation per minute(MV)was largely increased in the RSPF high dose group(P<0.05),as well as increased in the RSPF middle dose group and the positive group(P<0.01).?inflammatory cells in BALF:Compared with the model group,the number of inflammatory cells decreased significantly in the positive group(P<0.05)and in the RSPF middle and high dose groups(P<0.01).? lung coefficient and HYP content:Compared with the model group,the lung coefficient and HYP content of the rats decreased significantly in the positive group,and the RSPF low,middle and high dose groups(P<0.01).?lung histopathology:The alveolar structure in the model group was completely destroyed,with large areas of substantial changes and a large number of blue collagen deposition.Compared with the model group,the lung histopathological damage of rats in each group was alleviated,especially in the RSPF middle and high dose groups.?mRNA and protein expressions of TGF-(31:Compared with the model group,the mRNA expression of TGF-?1 in lung tissue decreased significantly in the RSPF middle dose group and the positive group(P<0.01);the protein expression of TGF-?1 decreased significantly in the positive group and RSPF middle and high dose groups(P<0.01).? EMT related mRNA and protein expressions:Compared with the model group,the mRNA expression of a-SMA in the lung tissue decreased significantly in the RSPF middle and high dose groups and the positive group(P<0.05);the protein expression of a-SMA decreased significantly in the RPSF middle dose group(P<0.01)and the high dose group(P<0.05).The mRNA expression of E-cadherin in the lung tissue significantly increased in the RSPF low,middle and high dose groups(P<0.05);the protein expression of E-cadherin in the lung tissue significantly increased in the RSPF low,middle and high dose groups(P<0.01).?autophagy related mRNA and protein expressions:Compared with the model group,the mRNA expression of lc3b in lung tissue increased significantly in the RSPF middle dose group(P<0.01),as well as in the RSPF high dose group and the positive group(P<0.05);the protein expression of lc3b in lung tissue increased significantly in the RSPF high dose group and positive group(P<0.05).The mRNA and protein expressions of beclin-1 in lung tissue increased significantly in the RSPF middle dose group(P<0.05).3.The effect of PSPF on HELF stimulated by TGF-?1 in vitro:Compared with the normal control group,the cell proliferation of TGFP-?1 treatment group raised significantly,mRNA and protein expressions of a-SMA were increased,mRNA and protein expressions of E-cadherin,lc3b and beclin-1 were decreased.Compared with the TGF-?1 treatment group,the cell proliferation was reduced when treated with different concentrations of RSPF,and the changes of above indicators were reversed in different concentrations of RSPF group as well.Conclusion:RSPF has the effect on pulmonary fibrosis rats through increasing rats survival rate,improving lung histopathology changes,reducing lung coefficient and HYP content,etc.RSPF could also inhibit HELF proliferation induced by TGF-?1 in vitro.The anti-pulmonary fibrosis mechanism of RSPF might be associated with up-regulating the protein expressions of lc3b and Beclin-1,inhibiting the EMT process of pulmonary epithelial cells.The above results provide scientific basis for RSPF in the treatment of pulmonary fibrosis on clinic.