Clinical Observation And Mechanism Of Honghua Injection Combined With Conventional Western Medicine In The Treatment Of Community-acquired Pneumonia(Blood Stasis Blocking Collateral Syndrome)

Objective1.A prospective,randomized,parallel controlled,double-blind trial was conducted to evaluate the clinical efficacy and safety of Honghua Injection combined with conventional treatment of Western medicine in the treatment of CAP(blood stasis blocking collateral syndrome);2.Based on network pharmacology,the mechanism of Honghua injection in the treatment of community-acquired pneumonia was further elucidated;3.Based on Sunluo-microcirculation theory,The mechanism of Honghua injection in the treatment of CAP(blood stasis blocking collateral syndrome)was preliminarlly discussed from microcirculation disorders and related inflammatory signal transduction pathways.Methods1.Clinical observation of Honghua injection combined with conventional treatment of Western medicine in the treatment of CAP(blood stasis blocking collateral syndrome)Sixty patients were selected from the Respiratory Department of the Affiliated Hospital of Chengdu University of Traditional Chinese Medicine from June 2018 to June 2019.Strict inclusion and exclusion criteria were formulated in this study.All 60 patients met the inclusion criteria.Procplan was used to randomly divide the patients into control group and treatment group,with 30 patients in each group,and opaque sealed envelope was used for allocation and concealment.The gender,age and other general conditions of the patients were statistically analyzed.The control group was given conventional Western medicine treatment,and the treatment group was combined with Honghua injection for 7 days.The clinical efficacy and safety of Honghua injection in the treatment of patients with CAP(blood stasis blocking collateral syndrome)were evaluated through clinical efficacy,chest CT absorption rate,improvement time of main symptoms and physical and chemical indexes,length of hospitalization,total TCM syndrome score,changes in TCM syndrome frequency,and incidence of adverse events;2.Based on network pharmacology,the mechanism of Honghua injection in the treatment of community-acquired pneumonia was further elucidatedThe main active components of Honghua injection were collected from literatures,and the SDF structure of the components was obtained by using Pub Chem database.The action targets of the components were obtained by introducing the SDF structure into Swiss Target Prediction database.CAP targets were obtained using OMIM,DISGENET and GENECARDS databases.The Network diagram of "component-target-disease" was constructed by Cytoscape3.7.2 software,and the main active components of Safflower injection were analyzed by the function of Network Analyzer.The PPI Network of protein interaction was constructed by the String database platform.The GO and KEGG enrichment analysis of key target genes was conducted based on R language and the Bioconductor platform.3.Discussion on the mechanism of Honghua injection in the treatment of CAP(blood stasis blocking collateral syndrome)Sixty cases of CAP(blood stasis blocking collateral syndrome)patients who met the inclusion criteria were selected in the Respiratory Department of Affiliated Hospital of Chengdu University of Traditional Chinese Medicine from June 2018 to June 2019,and 30 cases of healthy people who underwent physical examination in the same period were selected as the healthy group.Peripheral venous blood(7m L)was extracted on an empty stomach.One of the blood samples was placed in the red tube,and the supernatant was taken after separation,and the relevant indicators were detected by ELISA.The other was placed in the sterile EDTAK3 anticoagulant sampling tube for mononuclear cell separation and fluorescence quantitative PCR detection.Peripheral venous blood was extracted after 7 days of treatment(same method as above).Peripheral venous blood was extracted after 7 days of treatment(same method as above).Interleukin 1 beta(IL-1 beta),interleukin 6(IL-6),interleukin 8(IL-8),tumor necrosis factor alpha(TNF alpha)were chosen as inflammation evaluation indexes,NO and ET-1 were used as indicators of microcirculation disorder,TLR4 m RNA,p38 MAPKm RNA,AKT1 m RNA and nf-kappa Bm RNA were selected as index of cell signal transduction pathways,preliminarily discuss the mechanism of Honghua injection in the treatment of CAP(blood stasis resistance network card).Results:1.Clinical efficacy and safety of Honghua injection in the treatment of CAP(blood stasis blocking collateral syndrome): a randomized,parallel controlled,double-blind trial(1)Baseline level: There were no significant differences between the two groups in gender,mean age,total score of TCM syndrome,WBC,N%,ESR,CRP,PCT,D-D,PT,APTT,TT,FIB(P > 0.05),indicating comparability.(2)Clinical efficacy: After 7 days of treatment,the total effective rate of the treatment group was 93%,and the total effective rate of the control group was 68%.The clinical efficacy of the treatment group was better than that of the control group(P < 0.05).(3)Chest CT absorption rate: After 7 days of treatment,the chest CT absorption rate of the treatment group was better than that of the control group(P < 0.05);(4)Time of Symptoms disappeared,WBC improvement and length of hospital stay:there were statistically differences in the time of of cough、sputum chest pain disappearance,fever subsidence,WBC returning to normal and length of hospital stay(P < 0.05);(5)Total scores of TCM syndromes: After 7d of treatment,the improvement of total scores of TCM syndromes in the treatment group was better than that in the control group(P < 0.05);(6)Laboratory indicators: after 7 days of treatment,WBC,N%,CRP,PCTESR,DD,FIB values in treatment group were better than those in control group(P < 0.05);(7)Single item of TCM syndrome : After 7 days of treatment,the improvement of chest pain in the treatment group was significantly better than that in the control group(P < 0.01).The improvement of cough,sputum and sore throat in the treatment group was better than the control group(P < 0.05).There was no significant difference of dry mouth,thirsty between the two groups(P>0.05).(8)Safety analysis:6 adverse events occurred in 5 patients,3 adverse events occurred in 2 patients in the treatment group and 3 in the control group.Safety analysis of the two groups showed no significant difference in the incidence of adverse events between the two groups(P > 0.05).There were no serious adverse events in the two groups in this study,and the test drugs were safe and reliable,and the safety evaluation was level 2.2.To investigate the mechanism of Honghua injection in the treatment of CAP(blood stasis blocking collateral syndrome)11 potential active ingredients of safflower injection were screened,249 target points of component action,904 target points of CAP disease,and a total of 70 intersection targets.The main core targets include AKT1,TNF,EGFR,SRC,CXCL8,STAT3.The core target is related to anti-tumor,immune response,inflammatory response and so on.Enrichment analysis of 125 related pathways,mainly including proteoglycans in cancer;PI3K-Akt signaling pathway;EGFR tyrosine kinase inhibitor resistance;endocrine resistance;focal adhesions;human cytomegalovirus infection;Kaposi West’s sarcoma-related herpes virus infection;AGE-RAGE signaling pathway in the complications of diabetes,etc.3.To investigate the mechanism of Honghua injection in the treatment of CAP(blood stasis blocking collateral syndrome)(1)Baseline comparison of the three groups: there was no significant difference in gender and mean age between the three groups,and they were comparable(P>0.05).(2)ELISA detection of related factors: Before treatment,the levels of TNF-α,IL-1β,IL-6,IL-8 and ET-1 in patients with CAP(blood stasis blocking collateral syndrome)were significantly higher than those in healthy groups(P < 0.01).The concentration of NO was significantly lower than that of healthy group(P < 0.01);After treatment,the decrease of serum ET1,TNF-α,IL-8,IL-1β and IL-6concentration in treatment group was better than that in control group(P < 0.05),and the increase of serum NO concentration in treatment group was better than that in control group(P < 0.01).(3)TLR4m RNA,p38 MAPKm RNA,Akt1 m RNA and NF-κ Bm RNA in peripheral blood mononuclear cells were detected by fluorescence quantitative PCR:Before treatment,the expression levels of TLR4 m RNA,p38 MAPKm RNA,Akt1 m RNA and NF-κBm RNA in CAP patients(blood stasis blocking collateral syndrome)were significantly higher than those in healthy group(P < 0.01).After the course of treatment,the expression of TLR4 m RNA,p38 MAPKm RNA,AKT1 m RNA and NF-κBm RNA in patients with CAP patients(blood stasis blocking collateral syndrome)in the treatment group was better than that in the control group(P < 0.01).Conclusion:1.Compared with western medicine treatment alone,Honghua injection combined with conventional treatment in the treatment of CAP(blood stasis blocking collateral syndrome)can effectively control inflammatory indexes,improve clinical symptoms and shorten the length of hospital stay.The overall efficacy is good,with the effective rate of 93% and the safety evaluation is Grade 2.Therefore,the method of activating blood circulation and removing blood stasis is effective in the treatment of CAP and worthy of clinical promotion.In the future,more clinical and basic studies can be carried out for further investigation.2.Honghua injection can act on CAP through multi-component,multi-target and multi-pathway,and the representative target genes AKT1,TNF,EGFR and PI3K-Akt signaling pathway can be further explored in future experiments.3.The mechanism of Honghua injection in the treatment of CAP(blood stasis syndrome)is related to its reduction of serum concentrations of inflammatory factors TNF-α,IL6,IL-1β,IL-8 and ET-1/NO ratio in patients with pneumonia.In addition,Honghua injection could inhibit the expressions of TLR4 m RNA,p38 MAPKm RNA,AKT1 m RNA and NF-κ Bm RNA in monocytes.The anti-inflammatory effect of Honghua injection may be related to the regulation of p13K/Akt /NF-κ B signal transduction pathway.