Systemic Pharmacological Research And Experimental Validation Of Gegen Qinlian Decoction In The Treatment Of Irritable Bowel Syndrome

Irritable bowel syndrome is a chronic gastrointestinal disease,characterized by high abdominal pain,abdominal distention,defecation habits and stool consistency.It is particularly difficult in clinical treatment for long course and easy recurrence.GeGen QinLian decoction,one of the classic prescriptions,was used for the treatment of irritable bowel syndrome for thousands of years.However,it is difficult to further develop the true value of this prescription for its unclear mechanism.The aim of this study is to explore the specific mechanism of GeGen QinLian Decoction in the treatment of irritable bowel syndrome,and it is proved by the experiment of visceral highly sensitive rat model.It may provide a theoretical reference for Prescription Optimization and medical personnel medication in future.The study consists the following two part:1?The active components and corresponding targets of GeGen QinLian decoction were screened by Systems Pharmacology.489 chemical compositions of GGQLD were obtained from the database of Traditional Chinese Medicine,and 100 active components were met by the screening conditions(OB?33%,DL?0.18,Caco2?0.4).Eventually,30 candidate compounds were selected by a series of addition and deletion.It were found that the regulation of hormone level,immune anti-inflammatory,repair of vascular endothelial cells and kinase are closely related to the pathogenesis of irritable bowel syndrome by the analysis of the pharmacological activity of these compounds,which proved that these components had the potential for the treatment of IBS active components.109 predicted targets were obtained by reverse molecular docking technology on the basis of above candidate compounds.70 targets related to IBS were collected from the database and literature.Then,30 candidate targets were obtained by analyzing the relationship between protein and protein.12 classical signal pathways were obtained from candidate target clustering analysis.The relationship map of treatment and its functional analysis could be divided into four functional domains:nerve,immune,inflammation,and intestinal close connexin,referring to the relationship in pathways,which is in correspondence with the pathological characteristics of IBS.12 targets appearing in this pathway diagram were considered as importance for the treatment of IBS by GeGen QinLian decoction.TCM acted on IBS targets by making effect on the four functional domains indirectly.The molecular functions of 12 targets included protein kinase activity,ion channel gating,biomolecular binding and regulation of growth factors.The corresponding biological processes are emotional stress,signal conduction,hormone and neurotransmitter regulation,cell apoptosis,protein phosphorylation,and muscle and cardiopulmonary function of muscle blood pressure.Besides,emotional stress,signal transduction,hormones and neurotransmitter regulation are closely related to the symptoms of IBS.In addition,these targets are mainly distributed in the nerve functional plate,so we think the nerve functional plate is the key part of regulation.The network of "medicine-compound-target" was constructed to evaluate the network topology parameters of the targets.The parameters of PKA and MAOB were optimal in the target of the neural function plate.Eventually,the two targets are the key targets for treatment of IBS.In the pathway diagram,5-HT,5-HT3R and 5-HT4R,which are directly related to key targets,could be used to assist the verification of this conjecture.2.The conjecture was identified on the basis of visceral hypersensitivity rat model.The successful model was proved by two aspects of rat's general form(weight,appearance score)and intestinal sensation(Abdominal withdrawal reflex,minimum volume threshold).(compared with normal group,there were significant differences in model group,P<0.01).The protein level of the relevant targets was determined by ELISA and the mRNA expression of the related targets was detected by RT-PCR.The results of experiment showed that the protein levels of PKA and MAOB in the high dose group increased significantly,and the levels of 5-HT,5-HT3R and 5-HT4R protein decreased significantly(P<0.05),compared with the model group.The mRNA level of 5-HT4R increased significantly,and PKA,MAOB,and 5-HT3R mRNA levels were significantly decreased.It confirmed the previous conjecture that GeGen QinLian decoction may affect 5-HT metabolism and play a role in the treatment of IBS by acting on MAOB and PKA.The regulation on MAOB directly affected the level of 5-HT and indirectly affected the level of related receptors.Its relationship may be highly sensitive to "MAOB-5HT-HTR-visceral hypersensitivity",and PKA was activated to regulate the level of water channel protein and TRPV1.There are two ways:"PKA-water channel protein-visceral hypersensitivity" and "PKA phosphorylation-TRPV 1-visceral hypersensitivity".