Professor Miao Qing’s Experience In Applying Maxingshigan Decoction And Mechanism Of Modified Maxingshigan Decoction In Intervention Of Acute Lung Injury

Acute lung injury(ALI)is an acute diffuse inflammatory lung injury caused by various intrapulmonary and extrapulmonary factors,with severe hypoxemia and respiratory distress as the main clinical manifestations.ALI is a common clinical critical illness with complex pathogenesis,scarcity of treatments,and high mortality.Traditional Chinese medicine(TCM)has unique advantages in treating ALI,and it is particularly important to explore the basis and mechanism of TCM in treating acute lung injury.Professor Miao Qing,the supervisor,has rich experience in the diagnosis and treatment of respiratory diseases and proposes the treatment of ALI from both qi and blood-Modified Maxingshigan Decoction(MMD).This study summarizes the supervisor’s experience in using MMD to treat respiratory diseases and explores the mechanism of MMD in the treatment of ALI.While inheriting the academic experience of the supervisor,it provides a theoretical basis for MMD in the treatment of ALI.Objective1 To summarize Professor Miao Qing’s clinical experience in treating respiratory system diseases with MMD;2 To analyze the core target,key components,key biological processes and core signaling pathways of MMD in treating ALI by network pharmacology;3 To observe the effects of MMD on the degree of lung tissue injury,wet/dry weight ratio(W/D)value,bronchoalveolar lavage fluid(BALF)protein content,serum inflammatory factor content and PI3K/AKT signaling pathway by ALI rat model,and to explore the mechanism of MMD in treating ALI.Methods1 Through learning from teachers in outpatient department and ward,the author sorts out the clinical cases of the supervisor,summarizes the experience in treating respiratory system diseases with MMD,and attach the cases for supporting evidence.2 The active active components and targets of MMD were screened through the TCMSP database,and the active component-target interaction network map was constructed.TTD,GeneCards,OMIM,CTD,DisGeNET databases were searched to collect disease targets for ALI.The MMD active ingredient targets were intersected with ALI targets to obtain drug-disease targets,and PPI analysis of network targets was performed in the STRING database,network construction and analysis were performed in Cytoscape,and GO and KEGG analysis were performed in the DAVID database.3 ALI rat models were constructed using lipopolysaccharide,and 72 SD rats were randomly divided into 6 groups(blank group,model group,dexamethasone group,MMD high-dose group,MMD medium-dose group,and MMD low-dose group),with 12 rats in each group.Drug intervention for 5 days.The general condition of the rats was observed,and after the intervention,the blood,BALF,and lung tissue were collected.HE staining and microscopy were used to observe the pathomorphological changes of lung tissues in each group,and Smith score was performed for the degree of lung injury to detect W/D values,BALF protein content was determined by BCA,serum TNF-α,IL-6,and IL-1β expression levels were detected by ELISA,PI3K,p-PI3K,AKT,and p-AKT protein expression was detected by Western blot,and PI3K and AKT gene expression levels in lung tissues were detected by qRT-PCR.Results1 The supervisor believed that MMD had the effects of promoting lung and clearing heat,relieving asthma and cough,and treated various fever diseases with its combination of Qingdao Decoction,Xiaochaihu Decoction and Haoqin Qingdan Decoction;combined with Suying Dingchuan Pills,Dilong,Dioscorea nipponica and Zhisou Powder to treat various cough,cough and asthma,acute exacerbation of chronic obstructive pulmonary disease;and proposed to treat ALI from the treatment of qi and blood.2 Network pharmacology:a total of 148 effective active components of MMD were obtained,47 effective active components after weight removal,258 active component targets,1537 disease targets,and 165 drug-disease targets,of which 27 effective active components of MMD related to drug-disease targets.Quercetin,wogonin,kaempferol,β-sitosterol,naringenin,baicalein,and sesamin,may be the key active components of MMD in the treatment of ALI.There are 38 drug-disease core targets,of which AKT1,MAPK3,MYC,CASP3,TP53,EGFR,MAPK8,HIF1 A,JUN and TNF are key core targets.GO bioconcentration analysis showed 1933 biological processes,97 cellular components and 185 molecular functions,of which biological processes were highly correlated with hormone response,cytokine response,inorganic response,reactive oxygen species response and oxidative stress response,etc.and the gene groups involving cellular components were mainly concentrated in membrane rafts,membrane microstructure domains and transcriptional regulatory complexes;molecular functions were closely related to DNA transcription factor binding,transcription factor binding,kinase binding and oxidoreductase activity.177 ALI related pathways were obtained by KEGG pathway enrichment analysis,and it was speculated that the process of MMD in the treatment of ALI was related to cancer signaling pathways,lipids and atherosclerosis,PI3K/AKT signaling pathway,hepatitis B,interleukin 7 signaling pathway,TNF signaling pathway,cytomegalovirus infection,and small cell lung cancer.In the KEGG database enrichment analysis results,PI3K/AKT signaling pathway was an important pathway,and 15 out of 38 core targets were associated with PI3K/AKT signaling pathway.3 Animal experiments:(1)General conditions:the rats in the blank group had no special behavioral abnormalities;the rats in the other groups showed apathy,shortness of breath,shrugged hair,dim color,trembling all over,decreased activity,cyanosis of lips and four claws,and some rats showed fecal and urinary incontinence.On days 2-5 of drug administration,the mental status,activity,eating and drinking of rats in each group gradually improved,and the status of rats in each treatment group was significantly better than that in the model group.(2)Lung histopathology:compared with the blank group,abnormal lung structure in the model group,the alveolar wall thickened,the alveolar space and interstitial edema,inflammatory cell infiltration,and increased Smith score(P<0.05);compared with the model group,the lung histopathology of the MMD groups was improved to different extents,in which the alveolar and interstitial edema and inflammatory infiltration were significantly improved in the MMD high and medium dose groups;the Smith scores of the MMD groups decreased,including MMD high and medium dose group(P<0.05),and no statistical difference in MMD low dose group(P>0.05).(3)W/D value of lung tissue:Compared with the blank group,the W/D value of the model group was significantly higher(P<0.05),and compared with the model group,the W/D value of the MMD groups decreased(P<0.05).(4)BALF protein content:compared with the blank group,the BALF protein content in the model group was significantly increased(P<0.05),and compared with the model group,the BALF protein content in the MMD groups decreased(P<0.05).(5)Serum inflammatory factor levels:Compared with the blank group,the levels of TNF-α,IL-6,and IL-1β in the model group were significantly higher(P<0.05);compared with the model group,the levels of TNF-α,IL-6,and IL-1β in the MMD groups were significantly lower(P<0.05).(6)PI3K,p-PI3K,AKT,p-AKT protein expression results:compared with the blank group,the PI3K phosphorylation level and AKT phosphorylation level in the model group were significantly increased(P<0.05);compared with the model group,the PI3K and AKT phosphorylation levels in the MMD high-dose group were significantly decreased(P<0.05),and there was no statistical difference in PI3K and AKT in MMD medium and low dose groups(P>0.05).(7)PI3K and AKT gene expression:compared with the blank group,PI3K and AKT mRNA expression in the model group was increased(P<0.05),and compared with the model group,PI3K and AKT mRNA expression levels in the MMD groups were decreased to different extents,with statistical differences in MMD high and medium dose groups(P<0.05)and no statistical differences in the MMD low dose group(P>0.05).Conclusion1 MMD is a common prescription for the supervisor in the treatment of fever,cough and asthma,which has a good clinical effect.2 MMD may regulate the expression of PI3K/AKT signaling pathway by combining core targets(AKT 1,MAPK 3,naringenin,MAPK 3,MYC,TNF,EGF,VE GF A,TP 53,etc.),thus inhibiting inflammatory response,affecting oxidative stress,promoting angiogenesis,regulating immune response,and playing a role in the treatment of ALI.In this study,the specific effect of the treatment of MMD in ALI was explained at the level of molecular mechanism,providing ideas for further experimental validation.3 Experimental results:MMD has a significant protective effect on ALI rats,which can reduce the degree of lung tissue injury,improve pulmonary edema,reduce the levels of inflammatory cytokines TNF-α,IL-6 and IL-1β,and inhibit the expression of PI3K/AKT signaling pathway.