| Irritable Bowel Syndrome(IBS)was one of the common functional gastrointestinal disorders.IBS was primarily characterized by chronic abdominal pain or discomfort,and altered bowel habits in the absence of evidence of organic diseases.Accumulating evidences demonstrate that it was related to visceral hypersensitivity.Among these,visceral hypersensitivity could be thought as a biological marker of the syndrome.Thus,the aim of our present study was to investigate the effect of CZNCF on visceral hypersensitivity in rats and its underlying action mechanisms.1.Multiple Stress Protocol Rats were subjected to 21 consecutive days.2 random stressors,abrosia,fasting water,water avoidance stress,cold restraint stress and forced swimming stress.Then,using general state(appearance behavior score,weight changes),the gut feeling(pain threshold pressure,abdominal withdrawal reflex)to appraisal the model,compared with the normal group,there are significant differences(P<0.01)t,he results showed that the model was success.2.To investigate the effect of ChaiZhu Ningchang fang(CZNCF)on irritable bowel syndrome model rats of 5-HT signal system.The concentrations of 5-HT in rat serum;tryptophan hydroxylase 1,5-hydroxytryptamine transporter,5-hydroxytryptamine 3 receptors and 5-hydroxytryptamine 4 receptors levels in colon were determined through the ELISA kit while the expression of mRNA of TPH1,SERT,5HT3R and 5 HT4R was detected by RT-PCR method.Compared with the normal group,significant variance(P<0.05)in the semi-quantitative score,AWR score and minimum volume threshold were observed in model group.The concentrations of the 5-HT in serum,the level of TPH1,5-HT3R in colon were increased(P<0.01,P<0.05)while the level and the expression of mRNA of SERT and 5-HT4R in colon were decreased(P<0.05).In treatment groups,ChaiZhu Ningchang fang significantly decreased the concentrations of the 5-HT in serum,the level and the expression of mRNA of TPH1,5-HT3R in colon while increased the level and the expression of mRNA of SERT and 5-HT4R in colon(P<0.05).3.To investigate the effect of ChaiZhu Ningchang fang effective components(Paeoniflorin,Glycyrrhizic acid,Synephrine)on irritable bowel syndrome model rats of 5-hydroxytryptamine signal system.ELISA and Real-time quantitative PCR were used to measure the levels of protein and mRNA of 5-HT pathway.Paeoniflorin and Glycyrrhizic acid group,the symplectic Synephrine group,Paeoniflorin,Glycyrrhizic acid,Paeoniflorin and Symplectic Synephrine group can significantly improve visceral high sensitive model rats(weight,appearance,behavior score)general condition,relieve visceral high sensitivity(abdominal withdrawal reflex score,minimum capacity threshold).The effect of high dose group is better than the Glycyrrhizic acid and glycyrrhizin low dose group(P<0.05);Paeoniflorin and Glycyrrhizic acid and Paeoniflorin-Symplectic Synephrine groups inhibition model rats weight loss trend is superior to the single into group(P<0.01).Compared with normal group,model group 5-HT content in the colon of rats,5-HT conversion and 5-HT3R,TPH1 protein level and mRNA level increased significantly(P<0.05,P<0.01),the model group rats colon 5-HT4R and SERT protein level and mRNA level was significantly lower(P<0.05,P<0.01).Compared with model group,Paeoniflorin,Glycyrrhizic acid and Paeoniflorin-Synephrine group 5-HT content in the colon of rats,5-HT conversion and 5-HT3R,TPH1 protein and mRNA level decreased significantly(P<0.05,P<0.01);Paeoniflorin and Glycyrrhizic acid and Paeoniflorin-Synephrine group 5-HT4R and SERT protein levels in colon of rats and mRNA level was significantly increased(P<0.05),the effect is better than that of each single effective into groups.In summary,the present study explored treatment of CZNCF and its effective componentson on visceral hypersensitivity in rats,and studied the therapeutic effect which may be related to 5-HT signaling pathway.This study not only provided experimental evidence for clinical efficacy,but also clarified the mechanism of traditional Chinese medicine for the treatment of IBS. |
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