| IBS is the most common chronic disorder of gastrointestinal(GI)function characterized by continuous or remittent abdominal pain and is associated with altered bowel habits,diarrhea and constipation or both,and bloating.Chronic abdominal pain or discomfort relieved by defecation and associated with altered bowel habits are the mainstay in diagnosis.The disorder can not be explained by specific pathophysiologic mechanisms,or structural or known biochemical abnormalities.Valerianoj atamansi Jones has good therapeutic effects on the treatment of gastrointestinal disorders,such as indigestion,abdominal distension,pain and diarrhea.The preliminary studies showed that Valerianoj atamansi Jones and iridoid constituents of Valeriana jatamansi Jones could reduce the expression of serotonin in the colon,decrease the expression of 5-HT3 recepter,improve the abnormal gastrointestinal dysfunction and spiritual activity and the colon gastrointestinal hormones(such as SP,VIP,SS),reduce the number of EC and MC in colon tissue.The iridoid constituents of Valeriana ja tamansi Jones also made the rats’ colon longitudinal muscle cells relax and improved the visceral sensitivity in rats with IBS.The baldrinal is a monomer which is extracted from Valerianoj atamansi Jones.The structure of baldrinal is stable.Its abundant,biological activity and research value of baldrinal are high.Our preliminary studies showed that baldrinal could reduce the expression of serotonin in the serum and TPH1 mRNA in the colon.Based on the preliminary research,the effects of baldrinal on visceral sensitivity,mental activity,food intake,fecal water content and morphology were observed in our research in order to evaluate the treatment effects of baldrinal on IBS model animals.The effects of baldrinal on 5-HT pathway,levels of SP and leptin in serum,the PKCy、c-fos in the spinal cord,the NK1 receptor mRNA in the brainstem were observed to explore the possible central mechanisms of the treatment effects of baldrinal.The peripheral mechanism was researched by another team member.1 The treatment effects of baldrinal in IBS rats1.1 Influence of baldrinal on the visceral hypersensitivity in IBS ratsObjective To investigate the influences of baldrinal on the visceral hypersensitivity in IBS rats.Methods Male SD rats were randomly divided into 6 groups:control group,IBS model group,pinaverium group(25mg/kg)and baldrinal high,medium and low dose group(0.6,0.3,0.15mg/kg).The IBS rat model was established by using CAS and independent feeding.After the model was established,rats were given the drugs by intragastric administration for 37 days.The abdominal withdrawal reflex(AWR)pressure threshold was assessed by colorectal distension before and after the establishment of IBS model.Results Before the establishment of IBS model and on day 14 after the establishment of IBS model,the AWR showed no significantly statistical meaning in each group of rats(P>0.05).On day 27,compared with the normal group rats,the AWR pressure threshold in IBS rats reduced significantly(P<0.01).On day 41,the medium and low dose of baldrinal significantly increased the AWR pressure threshold in IBS rats(P<0.01).On day 55 and 65,the baldrinal significantly increased the AWR pressure threshold in IBS rats(P<0.01).Conclusion The CAS and independent feeding could increase the visceral hypersensitivity in IBS rats.The baldrinal could significantly increase the AWR pressure threshold in IBS rats.1.2 Influence of baldrinal on the mental activity in IBS ratsObjective To investigate the influences of baldrinal on the mental activity in IBS rats.Methods Male SD rats were randomly divided into 6 groups:control group,IBS model group,fluoxetine group(2.5mg/kg)and baldrinal high,medium and low dose group(0.6,0.3,0.15mg/kg).The IBS rat model was established by using CAS and independent feeding.After the model was established,rats were given the drugs by intragastric administration for 37 days.The Open-field test was observed in all groups before and after the establishment of IBS model.Results The Open-field test showed no significantly statistical meaning in each group of rats before the establishment of IBS model(P>0.05).On day 18 after the establishment of IBS model,the level and vertical scores reduced significantly in IBS rats(P<0.05 or P<0.01).On day 48,the high dose of baldrinal significantly increased the level scores(P<0.05),the high and medium dose of baldrinal significantly increased the vertical scores(P<0.05).On day 63,the high dose of baldrinal increased the level scores significantly(P<0.05),but the vertical scores had no significantly statistical meaning(P>0.05).Conclusion The CAS and independent feeding could reduce the level and vertical scores in IBS rats.The baldrinal could significantly increase the level scores in IBS rats.1.3 Influence of baldrinal on the fecal water content in IBS ratsObjective To investigate the influences of baldrinal on the fecal water content in IBS rats.Methods Male SD rats were randomly divided into 6 groups:control group,IBS model group,pinaverium group(25mg/kg)and baldrinal high,medium and low dose group(0.6,0.3,0.15mg/kg).The IBS rat model was established by using CAS and independent feeding.After the model was established,rats were given the drugs by intragastric administration for 37 days.The fecal water content was observed in all groups before and after the establishment of IBS model.Results The fecal water content showed no significantly statistical meaning in each group of rats before establish IBS model(P>0.05).On day 25 after the establishment of IBS model,the fecal water content in IBS rats increased significantly(P<0.05 or P<0.01).On day 54 and 68,the baldrinal could reduce the the fecal water content in IBS rats significantly(P<0.05 or P<0.01).Conclusion The CAS and independent feeding could increase the fecal water content in IBS rats.The baldrinal could significantly reduce the the fecal water content in IBS rats.1.4 Influence of baldrinal on the food intake in IBS ratsObjective To investigate the influence of baldrinal on food intake in IBS rats.Methods Male SD rats were randomly divided into 6 groups:control group,IBS model group,pinaverium group(25mg/kg)and baldrinal high,medium and low dose group(0.6,0.3,0.15mg/kg).The IBS rat model was established by using CAS and independent feeding.After the model was established,rats were given the drugs by intragastric administration for 37 days.The influence of baldrinal on food intake was observed in all groups before and after the establishment of IBS model.Results The food intake had no significantly statistical meaning in each group of rats before establish IBS model and on day 9 and 24 after the establishment of IBS model(P>0.05).On day 47,the CAS and independent feeding reduced food intake in IBS rats(P<0.01).The high and medium dose of baldrinal significantly increased the food intake in IBS rats(P<0.05 or P<0.01).Conclusion The food intake in IBS rats reduced,the baldrinal could significantly increase the food intake in IBS rats.1.5 Influence of baldrinal on the morphology of colon in IBS ratsObjective To investigate the influences of baldrinal on morphology of colon in IBS rats.Methods Male SD rats were randomly divided into 6 groups:control group,IBS model group,pinaverium group(25mg/kg)and baldrinal high,medium and low dose group(0.6,0.3,0.15mg/kg).The IBS rat model was established by using CAS and independent feeding.After the model was established,rats were given the drugs by intragastric administration for 37 days.The influence of baldrinal on the morphology of colon was observed by HE staining.Results According to the results of HE staining,we could find the mucosal edema and inflammatory cells occasionally,there was no morphological and pathological changes in all group rat colon.Conclusion The CAS and independent feeding did not change the morphology of colon.2 The mechanisms of baldrinal in IBS rats2.1 Influence of baldrinal on the levels of 5-HT/5-HIAA and the conversion rate of 5-HT in hypothalamus and cortex of IBS ratsObjective To investigate the influences of baldrinal on the levels of 5-HT/5-HIAA and the conversion rate of 5-HT in hypothalamus and cortex of IBS rats.Methods Male SD rats were randomly divided into 6 groups:control group,IBS model group,fluoxetine group(2.5mg/kg)and baldrinal high,medium and low dose group(0.6,0.3,0.15mg/kg).The IBS rat model was established by using CAS and independent feeding.After the model was established,rats were given the drugs by intragastric administration for 37 days.5-HT/5-HIAA measured by phase high performance liquid chromatography with electrochemical detection(HPLC-ECD).Results Compared with the normal group,the levels of 5-HT was lower,the levels of 5-HIAA and the conversion rate of 5-HT were increased significantly in hypothalamus and cortex in the IBS rats(P<0.05 or P<0.01).The baldrinal could significantly increased the levels of 5-HT and decreased the conversion rate of 5-HT and the levels of 5-HIAA in hypothalamus in IBS rats(P<0.05 or P<0.01).The baldrinal could significantly increase the levels of 5-HT and decrease the conversion rate of 5-HT in cortex in the IBS rats(P<0.05 or P<0.01),but the levels of 5-HIAA in every group didn’t show significant difference.The treatment effect of baldrinal on the mental activity in IBS rats related to regulating the expression of 5-HT in CNS.Conclusion The CAS and independent feeding could decrease the levels of 5-HT and increase the conversion rate of 5-HT in IBS rats,the baldrinal could significantly increase the levels of 5-HT and decrease the conversion rate of 5-HT in hypothalamus and cortex of IBS rats.2.2 Influence of baldrinal on the expression of 5-HT in the hippocampus of IBS ratsObjective To investigate the influences of baldrinal on the expression of 5-HT in the hippocampus of IBS rats.Methods Male SD rats were randomly divided into 6 groups:control group,IBS model group,fluoxetine group(2.5mg/kg)and baldrinal high,medium and low dose group(0.6,0.3,0.15mg/kg).The IBS rat model was established by using CAS and independent feeding.After the model was established,rats were given the drugs by intragastric administration for 37 days.The change of the 5-HT in the hippocampus detected by the immunohistochemistry.Results According to the results of immunohistochemistry,5-HT expressed in the cytoplasm of neurons.Compared with the normal group,5-HT in the hippocampus of IBS rats decreased significantly(P<0.05).The high dose of baldrinal could significantly increase the 5-HT in IBS rats(P<0.01).Conclusion The CAS and independent feeding decreased the levels of 5-HT in IBS rats,the baldrinal could significantly increase the 5-HT in hippocampus in IBS rats.2.3 Influence of baldrinal on the expression of 5-HT in the spinal cord of IBS rats Objective To investigate the influences of baldrinal on the expression of 5-HT in the spinal cord of IBS rats.Methods Male SD rats were randomly divided into 6 groups:control group,IBS model group,pinaverium group(25mg/kg)and baldrinal high,medium and low dose group(0.6,0.3,0.15mg/kg).The IBS rat model was established by using CAS and independent feeding.After the model was established,rats were given the drugs by intragastric administration for 37 days.The change of the 5-HT in the spinal cord was detected by the immunohistochemistry.Results According to the results of immunohistochemistry,5-HT positive nerve fibers was in the spinal cord dorsal horn Ⅰ and Ⅱ layer.The cytoplasmic positive was yellow brown.Compared with the normal group,the 5-HT in the spinal cord of IBS rats increased significantly(P<0.05).The high and low dose of baldrinal could significantly reduce the 5-HT in the spinal cord of IBS rats(P<0.05 or P<0.01).Conclusion The 5-HT in the spinal cord in IBS rats be higher than controls,the baldrinal could significantly reduce the 5-HT in spinal cord tissue of IBS rats.2.4 Influence of baldrinal on the levels of SP and OB in serum of IBS ratsObjective To investigate the influences of baldrinal on the levels of SP and OB in serum of IBS rats.Methods Male SD rats were randomly divided into 6 groups:control group,IBS model group,pinaverium group(25mg/kg)and baldrinal high,medium and low dose group(0.6,0.3,0.15mg/kg).The IBS rat model was established by using CAS and independent feeding.After the model was established,rats were given the drugs by intragastric administration for 37 days.Serum SP and OB level were measured by enzyme-linked immunosorbent assay method.Results The levels of SP in IBS rats was higher than controls,but there was no significantly statistical meaning in each group(P>0.05).The high and low dose of baldrinal could reduce the levels of SP in IBS rats(P<0.05).However,the levels of OB in IBS rats was lower than controls,there was no significantly statistical meaning in each group,too(P>0.05).The high and medium dose of baldrinal could increase the levels of OB in IBS rats.Conclusion The levels of SP in IBS rats was higher than controls,the baldrinal reduced the levels of SP in serum of IBS rats.The levels of OB in IBS rats was lower than controls.The baldrinal increased the levels of OB in serum of IBS rats.2.5 Influence of baldrinal on the expression of PKCγ、c-fos in the spinal cord of IBS ratsObjective To investigate the influences of baldrinal on the expression of Protein kinase Cγ(PKCγ)、c-fos in the spinal cord in IBS rats.Methods Male SD rats were randomly divided into 6 groups:control group,IBS model group,pinaverium group(25mg/kg)and baldrinal high,medium and low dose group(0.6,0.3,0.15mg/kg).The IBS rat model was established by using CAS and independent feeding.After the model was established,rats were given the drugs by intragastric administration for 37 days.The changes of the PKCγ and c-fos were detected by the immunohistochemistry.Results According to the results of immunohistochemistry,PKCγ positive cells concentrated in the spinal dorsal horn of shallow and looked like the circular or spindle.The positive immune reaction product distributed in the cytoplasm.The color is tan.c-fos positive cells concentrated in the spinal cord dorsal horn Ⅰ、Ⅱ and Ⅳlayer.The c-fos focused on neurons,the cytoplasmic positive is yellow to brown.The CAS and independent feeding could lead to the PKCγ and c-fos increased significantly(P<0.01).The baldrinal reduced the PKCγ in IBS rats significantly(P<0.01),the high and medium dose of baldrinal could significantly reduce the c-fos in IBS rats(P<0.05 or P<0.01).Conclusion The PKCγ and c-fos in the spinal cord in IBS rats were higher than controls.The baldrinal reduced the PKCγ and c-fos in spinal cord in IBS rats.2.6 Influence of baldrinal on the mRNA expression of NK1 receptor in brainstem of IBS ratsObjective To investigate the influences of baldrinal on the mRNA expression of neurokinin-1(NK1)receptor in brainstem of IBS rats.Methods Male SD rats were randomly divided into 6 groups:control group,IBS model group,pinaverium group(25mg/kg)and baldrinal high,medium and low dose group(0.6,0.3,0.15mg/kg).The IBS rat model was established by using CAS and independent feeding.After the model was established,rats were given the drugs by intragastric administration for 37 days.The mRNA expression of NK1 receptor was detected by using RT-PCR.Results The CAS and independent feeding increased the mRNA expression of NK1 receptor in IBS rats(P<0.01).The high and medium dose of baldrinal could significantly reduce the mRNA expression of NK1 receptor in IBS rats(P<0.05 or P<0.01).Conclusion The mRNA expression of NK1 receptor in IBS rats was higher than controls,the baldrinal could significantly reduce the mRNA expression of NK1 receptor in brain in IBS rats.We could get the following conclusions:1.The IBS rat model was established by using CAS and independent feeding.The IBS rats showed the following changes:visceral hypersensibility,abnormal mental activity,loss of appetite,diarrhea.These could simulation part of IBS symptoms.The baldrinal could significantly increase the AWR pressure threshold,regulate the mental activity and increase food intake.It could relieve abdominal pain and diarrhea in IBS rats.2.The expression of 5-HT in central nervous system(hippocampus,hypothalamus and cortex)of IBS rats reduced significantly.The baldrinal could significantly increase the levels of 5-HT and decrease the conversion rate of 5-HT in hypothalamus and cortex of IBS rats.The 5-HT in the CNS could exert analgesic effect and regulate the mental activity.3.The 5-HT、PKCγ、c-fos in spinal cord tissue of IBS rats increased significantly,they could enhance the transmission of nociceptive information and lead to the visceral hypersensibility.The baldrinal could significantly reduce the expression of 5-HT、PKCγ and c-fos in spinal cord tissue and relieve abdominal pain.4.The level of SP in serum and the mRNA expression of NK1 receptor increased in IBS rats.The combination of SP and NK1 receptor could speed up the bowel movement and attenuate the appetite.The levels of OB in IBS rats reduced significantly.The baldrinal could significantly reduce the level of SP and NK1 receptor,regulate the bowel movement and appetite and increase the levels of OB.5.The treatment effects of baldrinal in IBS rats were related to regulating the expression of 5-HT,NK1 receptor in CNS,5-HT、PKCγ and c-fos in spinal cord tissue,SP and leptin in serum. |
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