The Mechanism And Clinical Application Of Qingkailing Injection Based On Network Pharmacology And Meta-analysis

Qingkailing Injection has two varieties nowadays,Qingkailing Injecta and Qingkailing Freeze-dried Powder for Injection,both mainly consist of cholic acid,hyodeoxycholic acid,bubali cornu,baicalin,gardeniae fructus,lonicerae japonicae flos,isatidis radix,and margaritifera concha,the efficacy of which including heat-clearing,detoxicating,resoliving phlegm decoction,dredging collaterals and inducing resuscitation,having many kinds of pharmacological effects such as antibacterial,antiviral,antipyretic,anti-inflammatory,immunoregulation,liver protection,improving brain circulation and protecting the function of brain cell.Qingkailing Injection is widely applied in clinical treatment,and has good curative effect especially in cerebrovascular disease,the upper respiratory tract infection,hepatitis and high fever.The pharmacodyamic material basis,mechanism of action and clinical evaluation research of Qingkailing Injection has always been the hot spot of the academic research.Thus,based on the network pharmacology,by means of multilevel biological networks for herbs,compounds,targets and diseases of Qingkailing Injection,combining with gene functional annotation and KEGG pathway,we carried out research on the mechanism of action of Qingkailing Injection;taking advantage of evidence-based medicine methods,meta-analysis and system evaluation,we conducted scientific assessment of the methodological quality and relevant data of clinical randomized controlled trials(RCTs),producing evidence for clinical reference on Qingkailing Injection treatment of specific diseases.Objective1 To explore the mechanism of action of Qingkailing Injection,revealing the characteristics of pharmacological action of Qingkailing Injection from the microscopic view.2 To systematically evaluate the efficacy and safety of Qingkailing Injection in the treatment of acute upper respiratory tract infection,pneumonia and mumps.Methods1 Network pharmacologyChemical compounds and molecule information related to Qingkailing Injection were searched through public databases such as TCMSP,PubChem Compound and TCMID;active compounds were predicted and screened on the ground of ADME property parameters and literature research;targets,genes and diseases related to herbs were further obtained.The database was built using Microsoft Access 2010.After extracting data from the database,"herb-compound-target-disease" networks were built by Cytoscape 3.4.0 software;then,network topology characteristics analysis was conducted to identify hub nodes and key modules of the networks;further,GO functional enrichment analysis and KEGG pathway enrichment analysis were carried out to analyze the primary function of the genes sets.Thus,we could explore the mechanism of action of Qingkailing Injection from multiple levels and perspectives.2 Meta-analysisRCTs regarding Qingkailing Injection for the treatment of specific diseases were collected through comprehensive searches of many Chinese and English databases such as CNKI,WANFANG DATA,VIP,SinoMed,PubMed,Springer,Web of Science and Cochrane Central Register of Controlled Trials.Related RCTs were selected according to the inclusion and exclusion criteria.After extracting data and assessing the methodological quality of included studies,meta-analysis was developed among the included studies with good homogeneity,the results of which presented with forest plot.If there were big heterogeneity among the included studies,meta-regression,subgroup-analysis and sensitivity-analysis could be used to explore sources of heterogeneity;furthermore,the sensitivity-analysis could be used to test the stability of the results,and the funnel plot could be used to detect publication bias.ResultsThe mechanism research based on network pharmacology1 Database building:The "Qingkailing Injection Network Pharmacology Basic Information Database" contained 116 chemical compounds,1 271 targets and their genes,929 diseases and 3786 "compund-target-disease" interaction of 8 herbs.2 Disassembled prescription analysis of mechanism research of Qingkailing Injection:(1)The mechanism research of "cholic acid-hyodeoxycholic acid":The major chemical compounds cholic acid and hyodeoxycholic acid both had good drug-likeness.The"herb-compound-target-disease" network involved 6 targets and 8 diseases.The key targets were MR,LXRa and LXR?.The key diseases involved cerebral injury,atherosclerosis,and dyslipidemia,etc.GO biological processes mainly involved regulation of triglyceride biosynthetic process,regulation of cholesterol storage,regulation of cholesterol efflux and so on.KEGG pathways mainly involved insulin resistance and so on.(2)The mechanism research of "bubali cornu-margaritifera concha":6 chemical compounds generally had good drug-likeness.The "herb-compound-target-disease" network involved 765 targets and 595 diseases.The key targets included prostaglandin G/H synthase 2(COX-2)and peroxisome proliferator-activated receptor gamma(PPARy).Three diseases with the highest degree were schizophrenia,Alzheimer's disease and inflammation.GO biological processes mainly involved small molecule metabolic process,cell metabolic process and so on.KEGG pathways mainly involved metabolism pathway of various amino acids.(3)The mechanism research of "baicalin-gardeniae fructus":20 active chemical compounds generally had good drug-likeness.The "herb-compound-target-disease" network included 198 targets and 369 diseases.The key targets included COX-2,PPAR? and arachidonate 5-lipoxygenase(5-LOX).The key diseases primarily involved various cancers,diabetes,cardiovascular disease and so on.GO biological processes involved response to chemical stimulus,regulation of cell proliferation,regulation of programmed cell death and so on.The KEGG pathways mainly involved various cancer pathways and Hepatitis B.(4)The mechanism research of "lonicerae japonicae flos-isatidis radix":92 active chemical compounds generally had good drug-likeness.The "herb-compound-target-disease"network involved 665 targets and 609 diseases.The key targets included COX-2,heat shock protein HSP 90-alpha(HSP86),prostaglandin G/H synthase 1(COX-1)and PPARy.The key diseases primarily involved various cancers,cardiovascular disease,diabetes,Alzheimer's disease,inflammation and so on.GO biological processes involved cell surface receptor linked signaling pathway,response to chemical stimulus and so on.The primary KEGG pathways were cAMP signaling pathway,Calcium signaling pathway and various cancer pathways.3 Holistic analysis of mechanism research of Qingkailing Injection:116 active chemical compounds generally had good drug-likeness.The "herb-compound-target-disease" network involved 1 271 targets and 929 diseases.Three targets with the highest degree were COX-2,HSP86 and COX-1.The key diseases primarily involved various cancers,cardiovascular disease,diabetes,Alzheimer's disease,schizophrenia,inflammation and so on.GO biological processes primarily involved response to chemical stimulus,signal transmission,metabolic process and so on.The primary KEGG pathways were cAMP signaling pathway,cGMP-PKG signaling pathway,metabolism pathways concerning multiple amino acids,cancer pathways,and Hepatitis B.For module analysis,GO biological processes primarily involved regulation of heart rate,regulation of cardiac muscle contraction and so on.The primary KEGG pathways were adrenergic signaling in cardiomyocytes,cGMP-PKG signaling pathway and so on.For the 29 active chemical compounds with good blood-brain barrier permeability,GO biological processes primarily involved cholesterol metabolism process,cholesterol homeostasis and so on.The primary KEGG pathways were PPAR signaling pathway,synthesis pathways concerning various amino acids,Nicotine addiction,etc.Systematic review based on meta-analysis1 Qingkailing Injection for acute upper respiratory tract infection:42 RCTs involving 5 905 patients were included;"Qingkailing Injection(+ western medicine symptomatic treatment)" compared with "Ribavirin Injection(+ western medicine symptomatic treatment)"could improve the clinical total effective rate(RR = 1.19,95%Cl:1.16?1.22,P<0.000 01),etc.12 studies reported the occurrence of 42 cases adverse drug reactions/adverse drug events(ADRs/ADEs)in experimental groups and 11 studies reported the occurrence of 34 cases ADRs/ADEs in control groups.2 Qingkailing Injection for pneumonia:17 RCTs involving 1 702 patients were included;The Meta-analysis indicated that the combined use of Qingkailing Injection,antibiotics and western medicine symptomatic treatment could improve the clinical total effective rate(RR =1.23,950%CI:1.14?11.33,P<0.000 01),etc.4 studies reported the occurrence of ADRs/ADEs 27 cases in experimental groups and 64 cases in control groups,respectively.3 Qingkailing Injection for mumps:7 RCTs involving 580 patients were included;The Meta-analysis indicated that the combined use of Qingkailing Injection and western medicine symptomatic treatment could shorten the duration of fever(MD ?-1.16,950%CI:-1.36?-0.96,P<0.000 01),etc.ADRs/ADEs were not observed in 3 studies and were not mentioned in 4 studies.Conclusion1 The results of network hub nodes analysis,GO function annotation enrichment analysis and KEGG pathway enrichment analysis indicate that the mechanism of the four drug couples of Qingkailing Injection each has its own focus,which reflects the difference of efficacy among four drug couples and the mutual complement and reinforce among different mechanisms,embodying the meaning of compatibility of medicine and compound prescription in traditional Chinese medicine.2 The mechanism of action of Qingkailing Injection may be that,multiple components,by acting on key targets such as LXR,cox-2,HSP86 and PPARy,regulating multiple biological processes such as the cholesterol metabolism,cholesterol homeostasis,amino acid metabolism,cell proliferation and cell apoptosis,and participating in various pathways such as multiple signaling pathways,metabolism pathway and diseases pathway,play an effective role in the treatment of various diseases such as cancers,cardiovascular disease,diabetes,Alzheimer's disease,schizophrenia,inflammatory diseases,etc.3 Qingkailing Injection is mainly used for cerebrovascular disease in clinic,less used for treatment of cardiovascular disease,but our study find that the multiple targets and pathways Qingkailing Injection involving are closely associated with cardiovascular diseases such as myocardial infarction,which deserves further research.4 All of "herb-compound-target-disease" networks of Qingkailing Injection and its four drug couples have some scale-free and small-world network features,with low module properties.Thus,identification of hub nodes is a good method in its network analysis,and module analysis method can be chosen according to the specific situation.5 In clinical applications,compared with western medicine routine treatment or symptomatic treatment,combination of Qingkailing Injection can improve the effect of the treatment of acute upper respiratory tract infection,pneumonia and mumps.However,there was no exact conclusion concluded on safety based on the included studies.