| Irritable bowel syndrome(IBS)represents one of the most common gastrointestinal dis-orders,with the characters of abdominal pain,discomfort,associated with change of stool form and frequency.The pathogenesis of IBS may invole in gene polymorphism,altered mu-cosa permeability,gastrointestinal dysmotility,visceral hypersensitivity,psychology disturb-ance,brain-gut peptide secretion disorders and immune dysfunction.Altered gastrointestinal motility and visceral sensitivity are the primary pathophysiology features of IBS,and induce the IBS clinical symptoms including abdominal pain,diarrhea and constipation.Valerianoj atamansi Jones has good therapeutic effects on the treatment of various gas-trointestinal disorders,such as indigestion,abdominal distension,abdominal pain and diarrhea.Modern pharmacology studies showed that Valerianoj atamansi Jones has the pharmacologi-cal action like inhibitory action of the gastrointestinal motility,sedation,analgesia and anti-convulsant action.The preliminary studies showed that Valerianoj atamansi Jones and Iridoid constituents of Valeriana jatamansi Jones could effectively improve the abnormal gastrointes-tinal dysfunction and spiritual activity through adjusting the expression of serotonin(5-HT),colon gastrointestinal hormones and mast cells in rats with irritable bowel syndrome which was established by being exposed to chronic unpredictable stress.ZXX and ZXY are two monomers which are extracted from Valerianoj atamansi Jones.The structure of ZXX and ZXY are stable.The biological activity and research value of ZXX and ZXY are high.The preliminary research showed that ZXX and ZXY could alleviate diarrhea caused by reserpine and abdominal pain caused by acetic acid,suggesting that ZXX and ZXY could inhibit gas-trointestinal dynamics and analgesic.Whether ZXX and ZXY have therapeutic effects on IBS model animals have not been reported.Based on the preliminary research,the effects of ZXX and ZXY on visceral sensitivity,defecation in two hours,mental activity,food intake and morphology were observed in our research in order to evaluate their treatment effects on IBS model animals.The effects of ZXX and ZXY on 5-HT pathway,the expression of colon gastrointestinal hormone and MC were observed to explore the possible periphery mechanisms of the treatment effects of ZXX and ZXY.1 The treatment effects of ZXX and ZXY on IBS rats induced by chronic stressObjective:To observe the effects of ZXX and ZXY on visceral sensitivity,defecation,mental activity,food intake and morphology in IBS model rats.Methods:Male SD rats were randomly divided into control group,model group,pinav-erium group(25 mg/kg),Fluoxetine group(2.5 mg/kg),ZXX groups in three doses(1.2 mg/kg、0.6 mg/kg、0.3 mg/kg)and ZXY groups in three doses(0.6 mg/kg、0.3 mg/kg、0.15 mg/kg).The model was established by being exposed to chronic unpredictable stress.Animals of the treatment groups were intragastrically daily.Abdominal withdrawal reflex pressure threshold,dejection amounts in 2h,fecal water content,Open-field test behavior,the number of defecate in open-field,food intake in 24h and HE staining were observed in each group in order to evaluate the treatment effects of ZXX and ZXY.Results:Compared with the control group,the AWR pressure threshold,Open-field be-havior score and food intake of model group reduced significantly(P<0.01 or P<0.05),the dejection amounts,fecal water content and defecation grains in Open-field test of model group increased significantly(P<0.01).ZXX and ZXY could increase the pain pressure threshold and the toler-ated pressure threshold significantly(P<0.01).ZXX and ZXY could reduce the defecation grains(P<0.05 or P<0.01)excepting ZXY middle dose,ZXX middle dose and ZXY of high and low doses could reduce the moisture content of the model animal defecation(P<0.05 or P<0.01).ZXX and ZXY could increase the vertical scores of open-field test(P<0.01)except-ing ZXX small dose,but had no significant effect on the crossing scores(P>0.05),ZXX in middle and small doses and ZXY in middle dose could reduce the defecation grains in open field-test(P<0.05 or P<0.01).ZXY could increase the food intake of model animals(P<0.05 or P<0.01).According to the results of HE staining,there was no obviously pathological and morphological changes in each group of IBS rats colon.Conclusion:Long-term chronic stress led to increased visceral sensation,intestinal dy-namic hyperfunction,abnormal mental activity and reduced appetite in IBS rats.ZXX and ZXY could improve visceral sensation,abnormality of intestinal dynamics,mental activity and appetite.This indicated that ZXX and ZXY had therapeutic effect on IBS model rats caused by chronic stress2.The mechanisms of ZXX and ZXY on IBS rats induced by chronic stress2.1 Influences of ZXX and ZXY on the expression of periphery 5-HT and 5-HIAA in rats with IBS induced by chronic stressObjective:To investigate the effects of ZXX and ZXY on the levels of 5-HT and 5-HIAA of colon and serum in IBS rats.Methods:Male SD rats were randomly divided into control group,model group,pinav-erium group(25 mg/kg),Fluoxetine group(2.5 mg/kg),ZXX groups in three doses(1.2 mg/kg、0.6 mg/kg、0.3 mg/kg)and ZXY groups in three doses(0.6 mg/kg、0.3 mg/kg、0.15 mg/kg).The model was established by being exposed to chronic unpredictable stress.Animals of the treatment groups were intragastrically daily.High performance liquid chromatogram electro-chemical detection was employed to measure the levels of 5-HT and 5-HIAA of colon and se-rum in each group.Results:Compared with control group,the content of 5-HT and 5-HIAA in both colon and serum of model group increased significantly(P<0.05 or P<0.01),but the ratio of 5-HT/5-HIAA only increased slightly(P>0.05).Compared with model group,ZXX and ZXY could reduce the serum level of 5-HT(P<0.01),the large and middle doses of ZXX,the large and small doses of ZXY could reduce the serum level of 5-HIAA(P<0.05),ZXX in large dose and ZXY in middle dose could reduce 5-HT/5-HIAA ratio in serum(P<0.05).ZXY in middle and small doses could reduce the content of colon 5-HT(P<0.05 or P<0.01),ZXY in middle dose could reduce the level of 5-HIAA in colon(P<0.05),the small dose of ZXX and ZXY could reduce colon 5-HT/5-HIAA ratio(P<0.05 or P<0.01).Conclusion:ZXX and ZXY could reduce the levels of 5-HT,5-HIAA and 5-HT/5-HIAA ratio of colon and serum in IBS model rats.2.2 Influences of ZXX and ZXY on the activity of colon MAO-A in rats with IBS induced by chronic stressObjective:To investigate the effects of ZXX and ZXY on colon MAO-A activity of IBS rats induced by chronic stress.Methods:Male SD rats were randomly divided into control group,model group,pinav-erium group(25 mg/kg),Fluoxetine group(2.5 mg/kg),ZXX groups in three doses(1.2 mg/kg、0.6 mg/kg、0.3 mg/kg)and ZXY groups in three doses(0.6 mg/kg、0.3 mg/kg、0.15 mg/kg).The model was established by being exposed to chronic unpredictable stress.Animals of the treatment groups were intragastrically daily.Colorimetric method was used to determine the activity of MAO-A in each group.Results:Compared with control group,the activity of colon MAO-A in IBS model rats decreased significantly(P<0.05).Compared with model group,ZXX and ZXY in both middle and small doses could increased MAO-A activity in rat colon significantly(P<0.05 or P<0.01).Conclusion:ZXX,ZXY could reduce colon MAO-A activity in IBS rats.2.3 Influences of ZXX and ZXY on the expression of TPH1 mRNA in rats with IBS in-duced by chronic stressObjective:To investigate the effects of ZXX and ZXY on the expression of TPH1 mRNA in rats with IBS induced by chronic stress.Methods:Male SD rats were randomly divided into control group,model group,pinav-erium group(25 mg/kg),Fluoxetine group(2.5 mg/kg),ZXX groups in three doses(1.2 mg/kg、0.6 mg/kg、0.3 mg/kg)and ZXY groups in three doses(0.6 mg/kg、0.3 mg/kg、0.15 mg/kg).The model was established by being exposed to chronic unpredictable stress.Animals of the treatment groups were intragastrically daily.RT-PCR was used to probe the expression of TPH1 mRNA in colon in each group.Results:Compared with control group,the expression level of colon TPH1 mRNA in-creased significantly in model group(P<0.01).Compared with model group,ZXX in small dose and ZXY in middle and small doses could decrease the expression of colon TPH1 mRNA(P<0.05 or P<0.01).Conclusion:ZXX and ZXY could decrease colon TPH1 mRNA expression.2.4 Influences of ZXX and ZXY on the expression of colon SERT in rats with IBS in-duced by chronic stressObjective:To investigate the effects of ZXX and ZXY on the expression of colon SERT mRNA and protein in rats with IBS induced by chronic stress.Methods:Male SD rats were randomly divided into control group,model group,pinav-erium group(25 mg/kg),Fluoxetine group(2.5 mg/kg),ZXX groups in three doses(1.2 mg/kg、0.6 mg/kg、0.3 mg/kg)and ZXY groups in three doses(0.6 mg/kg、0.3 mg/kg、0.15 mg/kg).The model was established by being exposed to chronic unpredictable stress.Animals of the treatment groups were intragastrically daily.RT-PCR was used to detect the expression of SERT mRNA in colon in each group.Western-blot was used to detect the expression of SERT protein in colon in each group.Results:Compared with control group,the expression level of colon SERT mRNA re-duced significantly in IBS model rats(P<0.05),ZXX and ZXY had no significant effects on the SERT mRNA expression of the colon.The expression level of SERT protein in the colon of IBS model rats decreased slightly(P>0.05),ZXX in large dose and ZXY in middle dose could elevated the expression level of colon SERT protein,but there was no statistical difference.Conclusions:The expression level of colon SERT mRNA reduced significantly in IBS model rats,while there was no significant change on the expression level of colon SERT pro-tein.ZXX and ZXY had no significant effect on the expression of colon SERT.2.5 Influences of ZXX and ZXY on the expression of colon EC cell in rats with IBS in-duced by chronic stressObjective:To investigate the effects of ZXX and ZXY on the number of 5-HT immuno-reactive cells(EC)in IBS rats.Methods:Male SD rats were randomly divided into control group,model group,pinav-erium group(25 mg/kg),Fluoxetine group(2.5 mg/kg),ZXX groups in three doses(1.2 mg/kg、0.6 mg/kg、0.3 mg/kg)and ZXY groups in three doses(0.6 mg/kg、0.3 mg/kg、0.15 mg/kg).The model was established by being exposed to chronic unpredictable stress.Animals of the treatment groups were intragastrically daily.The immunohisto-chemistry was used to detecte the number of colon 5-HT immunoreactive cell(EC).Results:Compared with control group,the number of colon 5-HT immunoreactive cells(EC)increased significantly in IBS rats(P<0.01).Compared with model group,ZXX and ZXY in large doses could reduce the number of colon 5-HT immunoreactive cells(EC)(P<0.05).Conclusion:The number of colon 5-HT immunoreactive cells(EC)in IBS rats was signif-icantly increased,ZXX、ZXY could reduce the number of colon 5-HT immunoreactive cells(EC).2.6 Influences of ZXX and ZXY on the expression of colon 5-HT3A mRNA receptor in rats with IBS induced by chronic stressObjective:To investigate the effect of ZXX and ZXY on colon 5-HT3AR mRNA expres-sion in IBS rats.Methods:Male SD rats were randomly divided into control group,model group,pinav-erium group(25 mg/kg),Fluoxetine group(2.5 mg/kg),ZXX groups in three doses(1.2 mg/kg、0.6 mg/kg、0.3 mg/kg)and ZXY groups in three doses(0.6 mg/kg、0.3 mg/kg、0.15 mg/kg).The model was established by being exposed to chronic unpredictable stress.Animals of the treatment groups were intragastrically daily.The expression of 5-HT3AR mRNA in rat colon was detected by RT-PCR.Results:Compared with control group,IBS rats colon 5-HT3AR mRNA expression in-creased(P<0.01).Compared with model group,ZXY in small dose could significantly reduce colon 5-HT3AR mRNA expression level(P<0.05).Conclusion:5-HT3AR mRNA expression in IBS rat colon increased,ZXY could signifi-cantly reduce colon 5-HT3AR mRNA expression level.2.7 Influences of ZXX and ZXY on the expression of colon gastrointestinal hormones in rats with IBS induced by chronic stressObjective:To investigate the effect of ZXX and ZXY on the colon of VIP,SP and SS in IBS rats.Methods:Male SD rats were randomly divided into control group,model group,pinav-erium group(25 mg/kg),Fluoxetine group(2.5 mg/kg),ZXX groups in three doses(1.2 mg/kg、0.6 mg/kg、0.3 mg/kg)and ZXY groups in three doses(0.6 mg/kg、0.3 mg/kg、0.15 mg/kg).The model was established by being exposed to chronic unpredictable stress.Animals of the treatment groups were intragastrically daily.The radioimmunoassay was used to detecte the expression levels of colon VIP,SP and SS.Results:Compared with control group,the expression level of colon VIP increased in IBS rats(P<0.05),ZXX large and small doses,ZXY middle and small doses could reduce the expression level of colon VIP(P<0.05).There was no significant effect on colon SP and SS.Conclusion:Colon VIP level increased in model rats,ZXX、ZXY could reduce colon VIP level.2.8 Influences of ZXX and ZXY on the number of colon MC cell in rats with IBS induced by chronic stressObjective:To investigate the effect of ZXX and ZXY on the number of MC in IBS rats induced by chronic stress.Methods:Male SD rats were randomly divided into control group,model group,pinav-erium group(25 mg/kg),Fluoxetine group(2.5 mg/kg),ZXX groups in three doses(1.2 mg/kg、0.6 mg/kg、0.3 mg/kg)and ZXY groups in three doses(0.6 mg/kg、0.3 mg/kg、0.15 mg/kg).The model was established by being exposed to chronic unpredictable stress.Animals of the treatment groups were intragastrically daily while modeling.The number of colon MC in each group was detected with toluidine blue stain.Results:Compared with control group,the number of colon MC in IBS rats increased sig-nificantly(P<0.05).Compared with model group,ZXX、ZXY in large doses could reduce the number of colon MC in IBS rats(P<0.05).Conclusion:ZXX and ZXY could decrease the number of mast cells in IBS rats.Base on the above research,we can draw the following conclusions:1 After being exposed to chronic stress for two month,the IBS model rats showed the following changes:visceral hypersensibility,gastrointestinal hyperfunction,abnormal mental activity,loss of appetite.These could simulation part of D-IBS symptoms.2 ZXX,ZXY could improve AWR threshold pressure,reduce the grain number and def-ecate content,regulate open-field scores,reduce the open-field grain number and increase food intake which could effectively relieve abdominal pain and diarrhea in IBS rats.3 The levels of 5-HT and 5-HIAA in peripheral,the number of EC which secrete 5-HT and the expression level of colon TPH1 mRNA in IBS rats increased,which showed the syn-thesis of 5-HT enhanced.While the expression of colon SERT mRNA and MAO-A activity reduced,which showed the reuptake and degradation of 5-HT weaken.5-HT synthesis and metabolism imbalance was the cause of the increased of peripheral 5-HT content.ZXX and ZXY could reduce the synthesis of 5-HT,increase the degradation of 5-HT,reduce the expres-sion level of peripheral 5-HT,but had no significant influence on 5-HT reuptake.4 The expression level of colon 5-HT3AR mRNA in IBS model rats increased,ZXY could reduce 5-HT3AR mRNA expression.5 The expression of colon VIP in IBS rats increased,ZXX and ZXY could reduce colon VIP expression.6 The number of colon MC in IBS rats increased,ZXX and ZXY could reduce the num-ber of MC in colon,make peripheral 5-HT level returned to normal.7 The treatment effects of ZXX and ZXY on D-IBS model rats were related to the regu-lating of peripheral 5-HT pathways. |
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