Study On The Synergistic Regulation Of The Active Components Of Sini Powder On The Disease Network Of Irritable Bowel Syndrome

Si Ni San was the famous prescription recorded in "Treatise on Febrile Diseases and Miscellaneous Diseases" compiled by Zhang Zhongjing in the Han Dynasty.It's currently used for the clinical treatment of irritable bowel syndrome(IBS).However,because of the large number of active ingredients and the pharmacological mechanisms of clinical treatment of diseases were not clear,which hindered the discovery of clinical effective material basis.This research mainly explored the effective ingredients of Si Ni San and its coordinated regulation mechanisms of IBS via a comprehensive strategy including bioinformatics and systems pharmacology.Through bioinformatics and systems pharmacology methods,63 IBS genes were mainly involved in 3 important pathways(Neuroactive ligand-receptor interaction,Inflammatory mediator regulation of TRP channels and Serotonergic synapse);4therapeutic modules(mental psychology,immune inflammation,neuroendocrine and cross-talk modules);9 key molecules(ADRA2A,HTR2 A,F2RL1,F2RL3,TRPV1,PKC,PKA,IL-1B and NGF).The occurrence and development of IBS were revealed;the coordinated regulation mechanisms of the effective ingredients(synephrine,paeoniflorin and naringin)at these pathways,therapeutic modules and the key molecules levels were clarified.The effects of the AWR minimum capacity thresholds and general state scores of the visceral hypersensitivity model rats were researched through the effective ingredients synephrine,paeoniflorin and naringin and their combinations regulated.Compared with the model group,the AWR minimum capacity thresholds and general state scores of rats in each administration group were significantly improved(p<0.05,p<0.01,p<0.001).Intraclass correlation coefficient(ICC)results showed that the effective ingredient combinations were highly consistent with the control group(ICC= 0.687 ? 0.872),better than the single ingredient groups(ICC < 0.4).The effects of synephrine,paeoniflorin,naringin and their combinations on the m RNA and protein expression levels of the key molecules in the colon tissues of VHS rats were determined by real-time fluorescent quantitative PCR and western blotting techniques.The results showed that,compared with the control group,the m RNA expressions of ADRA2 A,TRPV1,F2RL3,F2RL1,PKC,PKA and the protein expressions of F2RL1,IL-1B,and PKC were significantly up-regulated in the model group(p<0.05,p<0.01,p<0.001),while the m RNA and protein levels of HTR2 A and NGF were significantly down-regulated(p<0.01,p<0.001).Compared with the model group,the ingredient combination groups regulated the m RNA expressions of F2RL1,TRPV1,HTR2 A,ADRA2A,F2RL3,PKA,PKC,NGF and the protein expressions of PKC,F2RL1 and IL-1B(p<0.05,p<0.01,p<0.001).While the synephrine +paeoniflorin + naringin group had improvement effects on the abnormal expressions of the key molecules(p<0.05,p<0.01,p<0.001),which was like the Si Ni San group.Compared with the single ingredient group,the ingredient combinations had a synergistic effect in regulating the mental psychology,immune inflammation,neuroendocrine and cross-talk modules of VHS rats.The multi-target coordinated effects of the effective ingredients on the m RNA expressions of the key molecules were evaluated through the principal component analysis(PCA)and similarity calculation.PCA results showed that there was a clear classification trend between the different groups.The single ingredient groups and the model group were poorly distinguishable,and there were obvious differences between the ingredient combinations group and the model group.Compared with the single ingredient groups,the ingredient combination groups had a higher degree of aggregation at various points,which were obviously tending to the control group.The synephrine + paeoniflorin + naringin group had a significant correction effect,which had a similar trend to the Si Ni San group.Similarity analysis results showed that the model group,paeoniflorin group and naringin group had similar similarity(0.046 ?0.088),the pinaverium bromide group,Si Ni San group,synephrine + paeoniflorin group and paeoniflorin + naringin group had moderate similarity(0.106 ? 0.151),the control group,synephrine + naringin group,synephrine + paeoniflorin + naringin group had strong similarity(0.165 ? 1.434).It showed that the effective ingredient combination groups were like the pinaverium bromide group and the Si Ni San group,and had a better multi-target correction effect on the key molecules.In summary,synephrine,paeoniflorin and naringin could effectively improve visceral hypersensitivity by regulating the key molecules of important pathways and therapeutic modules,and had a good prospect for multi-target treatment of IBS.