Effect And Mechanism Of KangTai Capsule On The Model Of Visceral Hypersensitivity Rats

Irritable bowel syndrome(IBS)is a functional bowel disorder characterized by visceral hypersensitivity and altered bowel motility.Recent studies indicated that the pathogeny of IBS was complex and was not clear.At present,it was considered that IBS involved with multi-factors,including mental factor,immune factor and social factor,et al. Due to be short of ideal therapy and drug,symptomatic treatment is the primary treatment in Western medicine at present.Multi-target,integrity effect and few side effects are superiority of traditional Chinese medicine(TCM).Stagnation of liver-energy and deficicy of spleen,imbalance of heart and stomach was one of the most common types and was treated by the therapy of comfortable liver strengthen spleen,comfortable stomach and quiet mental.The KangTai Capsule is frequently used in PiWei institute to treat IBS,special diarrhea-IBS and is confirmed safe in clinic.Visceral hypersensitivity is one of main character in IBS and is the main reason leading to pain and imbalance in stomach and intestine movements.It showed the threshold change induced the colon distention.In clinic the patients is easy to feel pain when colon distention and in experiment it showed that the pain threshold pressure decreased in rats.To investigate the effect and mechanism of KangTai Capsule,two kinds of model of visceral hypersensitivity models in rats were established.On the basis of visceral hypersensitivity models,the express of 5-HT,NO,SP,CGRP and 5-HT receptor and CRF receptor were measured after the treatment of KangTai Capsule.The investigation could not only provide experimental proof of guiding clinical medication and developing new medicine,but also conduce to study the treatment of IBS with TCM..1 Effect of KangTai Capsule on two kinds of visceral hypersensitivity models in ratsThe KangTai Capsule is frequently used in PiWei institute to treat IBS,special diarrhea-IBS and is confirmed safe in clinic.In order to confirm the effect of KangTai Capsule,two models of visceral hypersensitivity(VH)in rats were replicated. One models of VH is induced by given acetic acid in rectum every day for continuous 14 days between postnatal 8d and 21d.Then the threshold of abdominal withdrawal reflex was measured at 6w and 8w with rectal distension.The rats were oral given KangTai Capsule for continuous 14 days.It showed that the pressure threshold of abdomen lifting and body arching in model group showed a obvious decrease compared with the normal group(P＜0.01).Colon irritation in neonatal rats leaded to a state of visceral hypersensitivity to adult rats.The pressure threshold of abdomen lifting and body arching in KangTai Capsule with low,middle and high dose group showed a obvious increase compared with in model group(P＜0.01).And the pressure threshold in high dose group is close to the norm group.The results indicated that the KangTai Capsule have the good effect on the acetic acid-induced VH in rats.The other is induced by Separation from mother and baby every day between postnatal 2d and 14d.Then the threshold of abdominal withdrawal reflex was measured at 6w and 8w with rectal distension.The rats were oral given KangTai Capsule for continuous 14 days.It showed that the pressure threshold of abdomen lifting and body arching in model group showed a obvious decrease compared with the normal group(P＜0.01).Colon irritation in neonatal rats leaded to a state of visceral hypersensitivity to adult rats.The pressure threshold of abdomen lifting and body arching in KangTai Capsule with low,middle and high dose group showed a obvious increase compared with in model group(P＜0.01).And the pressure threshold in high dose group is close to the norm group.The results indicated that the KangTai Capsule have the good effect on the Separation -induced VH in rats.On the basis of Separation -induced VH in rats,the rats were forced the water avodivance stress(WAS)for one hour.The number of soil and the threshold of abdominal withdrawal reflex was measured.It showed that the pressure threshold of abdomen lifting and body arching in model group showed a obvious decrease compared with the normal group(P＜0.01).The pressure threshold of abdomen lifting and body arching in KangTai Capsule group with low,middle and high dose showed a obvious increase compared with in model group(P＜0.01).The number of soil in model group showed a obvious increase compared with the normal group(P＜0.01)and it in KangTai Capsule with low,middle and high dose group showed a obvious decrease compared with in model group(P＜0.01). The results indicated that the KangTal Capsule have the regulation effect on disorder of colon movement and visceral hypersensitivity.2 Effect of KangTai Capsule on NO and 5-HT in two kinds of model of visceral hypersensitivity ratsThe pathogeny of IBS was complex and was not clear.The 5-HT and NO in the colon involved in the formation of visceral hypersensitivity.NO is the important matter which can relax the muscle and inhibit the sense and peristalsis of enteric tract.5-HT in blood,enteric tract and brain is the important medium which can regulate the sense and movement in the nerve centre and enteric tract.After treatment of KangTai Capsule,the threshold pressure of rectal distension was measured.Then the content of NO in blood serum and the content of 5-HT in blood serum,brain and colon were observed in the experiment.In the acetic acid-induced VH rat,the content of NO in blood serum in the model group decreased compared with the normal group(P＜0.05).KangTai Capsule with low,middle and high dose obvious increased the content of NO compared with the model group (P＜0.05 or P＜0.01).The content of 5-HT in the model group was higher than it in the normal group(P＜0.05).KangTai Capsule with low,middle and high dose obvious increased the content of 5-HT compared with the model group(P＜0.05 or P＜0.01). Experiment study confirmed:With the effects of KangTai Capsule,the content of NO in blood serum was increased and the content of 5-HT in blood and colon decreased.The content of 5-HT in brain could be decreased to a degree.The KangTai Capsule showed affirmative curative effect on treating rats of visceral hypersensitivity.It could be one of the mechanisms of action to influence the content of NO and 5-HT.In the Separation -induced VH rat,the content of NO in blood serum in the model group decreased compared with the normal group(P＜0.05).KangTai Capsule with low,middle and high dose obvious increased the content of NO compared with the model group (P＜0.05 or P＜0.01).The content of 5-HT in the model group was higher than it in the normal group(P＜0.05).KangTai Capsule with low,middle and high dose obvious increased the content of 5-HT compared with the model group(P＜0.05 or P＜0.01).With the effects of KangTai Capsule,the content of NO in blood serum was increased and the content of 5-HT in blood and colon decreased.The content of 5-HT in brain could be decreased to a degree.The KangTai Capsule showed affirmative curative effect on treating rats of visceral hypersensitivity.It could be one of the mechanisms of action to influence the content of NO and 5-HT.3 Effect of KangTai Capsule on expression of 5-HT,SP,CGRP and CRF in colonic mucosa of visceral hypersensitivity model ratsRecently P substance,CGRP,CRF in the enteric tract became heat hot in research.P substance,CGRP,CRF in the enteric tract induced the visceral hypersensitivity by direct stimulating the primary sense fibre.CRF in the enteric tract also can produce the visceral hypersensitivity.To investigate the machismo of KangTai Capsule effect on visceral hypersensitivity,the expression of 5-HT,SP,CGRP and CRF in colonic mucosa was detected with immunohistochemical method.The tissue sections of colon were stained with immunohistochemical staining.In the acetic acid-induced VH rat,positive rates of 5-HT in the model group increased compared with the normal group(P＜0.05).KangTai Capsule with middle and hige dose group obvious decreased compared with the model group(P＜0.05 or P＜0.01).The positive rates of P substance in the model group was higher than it in the normal group(P＜0.05). KangTai Capsule with low,middle and high dose obvious decreased it compared with the model group(P＜0.05 or P＜0.01).The positive rates of CGRP in the model group increased compared with the normal group(P＜0.05).KangTai Capsule with low middle and hige dose obvious decreased compared with the model group(P＜0.05 or P＜0.01).Experiment study confirmed:The positive rates of 5-HT,SP and CGRP in colonic mucosa was significantly reduced after the administration of KangTai Capsule.In the Separation induced VH rat,the positive rates of 5-HT in the model group increased compared with the normal group(P＜0.05).KangTai Capsule with middle and hige dose obvious decreased compared with the model group(P＜0.05 or P＜0.01).The positive rates of P substance in the model group was higher than it in the normal group (P＜0.05).KangTai Capsule with low,middle and high dose obvious decreased it compared with the model group(P＜0.05or P＜0.01).The positive rates of CGRP in the model group increased compared with the normal group(P＜0.05).KangTai Capsule with low middle and hige dose obvious decreased compared with the model group(P＜0.05or P＜0.01).Experiment study confirmed:The positive rates of 5-HT and CRF in colonic mucosa was significantly reduced after the administration of KangTai Capsule.It could be one of the mechanisms of action to regulate the express of 5-HT,SP,CGRP,CRF.4 Effect of KangTai Capsule on gene expression of 5-HT₃,5-HT₄ and CRF₁ in colonic mucosa of visceral hypersensitivity model ratsIt was known that 5-HT₃ receptor and 5-HT₄ receptor involved in the formation of visceral hypersensitivity.CRF₁ receptor in the brain and colon is related to the stress-induced visceral hypersensitivity.To investigate the mechanism of KangTai Capsule effect on visceral hypersensitivity model rats,the total RNA was isolated from these rats colonic mucosa.RT-PCR was used to detect 5-HT₃,5-HT₄ and CRF₁ expression andβ-actin was used as internal index.The products of PCR were separated with gel electrophoresis and stained.To compare the relative expression which was calculated from the gray scale ratio of 5-HT₃,5-HT₄ andβ-actin as well 5-HT₃,CRF₁ andβ-actin.In the acetic acid-induced VH rat,relative gene expression of 5-HT₃ in model group were significantly higher than that in normal group(P＜0.05).And relative expression of 5-HT₄ in model group were significantly lower than that in normal group(P＜0.05).Relative gene expression of 5-HT₃ in KangTai Capsule group with middle and high dose were significantly lower than that in model group(P＜0.05).And relative expression of 5-HT₄ in KangTai Capsule with middle and high dose group were significantly higher than that in model group(P＜0.05).In the Separation-induced VH rat,results in the model of chronic visceral hypersensitivity showed that relative gene expression of 5-HT₃ and CRF₁ in model group were significantly higher than that in normal group(P＜0.05).Relative gene expression of 5-HT₃ and CRF₁ in KangTai Capsule group middle and high dose were significantly lower than that in model group(P＜0.05).It indicated that the KangTai Capsule have the regulation effect on the express of 5-HT₃,5-HT₄ and CRF₁ receptor in colon.5 ConclusionIn the acetic acid-induced and Separation-induced VH rat models,the express of 5-HT,SP and CGRP in the colon was significantly higher than that in normal group.Also the express of 5-HT₃,CRF₁ receptor increased and the express of 5-HT₄ receptor dereased compared with normal group.It indicated that KangTai Capsule have the regulation effect on the express of 5-HT₃,5-HT₄ and CRF₁ receptor in colon.These may be one to the mechanisms of action to influence the VH.It also showed that the therapy of comfortable liver strengthen spleen,comfortable stomach and quiet mental are useful to explain the Multi-target,integrity effect of Chinese herbs.