| Irritable bowel syndrome(IBS) is a common functional intestinal disorderin clinic which characterized by abdominal pain and distension, associatedwith altered bowel habits and abdominal discomfort, includingdiarrhea-predominant, constipation-predominant, mixed and unclassified,diarrhea-predominant is the most common subtype. The prevalence is high.IBS not only affects the patient’s life quality and work efficiency, but alsoexpends lots of public health resources.Its basis of pathophysiology is mainlybased on the abnormal of gastrointestinal kinetics and the increased visceralsensitivity, but the mechanism is not clear,may be related to changes ofgastrointestinal motility, visceral hypersensitivity, brain-gut axis dysfunction,emotional disturbance, immune system dysfunction and endocrine abnormality.After studying thoroughly these years,brain-gut axis takes an important part inthe pathogenesis of IBS.In brain-gut axis, exists a class of small molecularpolypeptids, having dual function of hormones and neurotransmitters whichnamed as gut-brain peptide, plays a very important role in the regulation ofvisceral sensations,secretion and movement.Chinese medicine has its own superiority and characteristic in treatingIBS and has shown a widely application prospect. Jianpi Huashigranule(JPHSG) which was constituted of ten kinds of Chinese medicines,such as Radix astragali preparata,Fructus alpinae oxphyllae, Rhizomaatractylodis macrocephalae, Poria and Fructus amomi, made by moderntechnology, with the effects of regulate qi, invigorate spleen, resolve dampnessand antidiarrhea. In this study, Diarrhea-predominant Irritable BowelSyndrome(D-IBS) was induced by administration of Sennae decoctioncombined with restraint stress, and interfered with different doses of Jianpi Huashi granule. Study its effects on colon movement and visceral sensitivityof D-IBS model rat. Investigate the mechanism from the changes in the centralnervous system of c-fos, Corticotropin-releasing factor(CRF),Corticotropin-releasing factor receptor1(CRF-R1) and5-hydroxytryptamine(5-HT), so as to give theoretical basis for the treatment ofD-IBS.Part1The therapeutic effect of Jianpi Huashi granule on D-IBS model ratObjective:To study the effects of Jianpi Huashi granule on colon motility andvisceral sensitivity of D-IBS model rat, and evaluate its therapeutic efficacy.Methods:1Animals grouping and modelingSeventy-two male Sprague-Dawley rats were equally and randomlydivided into six groups:normal control group(NC), model group(M),Low-dose group of Jianpi Huashi Granule(JPHSG-L), Middle-dose group ofJianpi Huashi Granule(JPHSG-M), High-dose group of Jianpi HuashiGranule(JPHSG-H) and positive control group(PC). Except NC group, the restgroups were inducing of D-IBS by administration of Sennae decoctioncombined with restraint stress.2Medication and treatmentAfter modeling, each treatment group was given corresponding medicineby intragastric administration. NC and M groups were given sodium chloridewith equal dosage, all for14days.3The Therapeutic Effect observationSigning the body weight, using4-channel physiological recorder tracingsof colonic motility curve, and abdominal withdrawal reflex(AWR) evaluatingthe visceral sensitivity before and7,14days after D-IBS, and before and7,14days after treatment.4Colonic tissue pathology observationThe rats were sacrificed, and the distal colon to do HE staining. observed under microscope, elimination of the intestinal mucosa lesions.Results:1The effects of JPHSG on the weight of D-IBS Model RatBefore modeling, there were no significant difference among differentgroups(P>0.05); After modeling,the body weight was slowly increased, havingsignificant difference compared with NC group(P<0.01); After drugintervention, when compared with M group, the body weight gains weresignificantly higher in JPHSG-H group(P<0.05), and no significant differencein other treatment groups(P>0.05).2The effects of JPHSG on the colon motility function of D-IBS ModelRatBefore modeling, there were no significant difference among differentgroups(P>0.05); After modeling, the colonic motility index and the changerate of motility index were increased, having significant difference comparedwith NC group(P<0.01); After drug intervention, when compared with Mgroup, JPHSG-M and JPHSG-H groups were significantly decreased(P<0.01),compared with PC group, JPHSG-H group were significantlydecreased(P<0.05).3The effects of JPHSG on the visceral sensitivity of D-IBS Model RatBefore modeling, there were no significant difference among differentgroups(P>0.05); After modeling,when the score of AWR was3showed asignificant decrease, having significant difference compared with NC group(P<0.01); After drug intervention, when compared with M group, JPHSG-Mand JPHSG-H groups were significantly increased(P<0.01), compared withPC group, JPHSG-H group was significantly increased(P<0.05).4Colonic tissue pathology observationObservation of colon and microscopic tissue staining. Each groupshowed no edema,ulcer, congestion, inflammatory cell infiltration and so on,Compared with NC group, there were no significant difference in othergroups. Conclusions:JPHSG have commendable therapeutical effect on D-IBS model rats,improved the colon movement state and reduced visceral sensitivity.Part2The Central Mechanism Research of Jianpi Huashi Granule onDiarrhea-predominant Irritable Bowel SyndromeObjective:To study the effects of JPHSG on c-fos/CRF/CRF-R1/5-HT expression incentral of D-IBS model rat, and to investigate its mechanism of action.Methods:1Collection and processingAfter treated for14days, all the rats were anaesthetized and rapidlyretrieved the spinal dorsal horn,prefrontal cortex,hippocampus andhypothalamus, some fixed into the4%paraformaldehyde forimmunohistochemistry assay, and other stored at-80℃for ELISA and PCRassay.2Immunohistochemistry assay was used to detect the expression ofc-fos/CRF/CRF-R1/5-HT in central,calculate its mean density,and analyzed inhalf-ration.3ELISA was used to detect the content of c-fos and5HT in central of rat4PCR was used to detect the expression of CRF and CRF-R1mRNA incentral of rat, calculate its relative optical density(ROD), and analyzed inhalf-ration.Results:1The effects of JPHSG on c-fos/CRF/CRF-R1/5-HT expression in centralof D-IBS Model RatObserved under light microscope,positive stainings ofc-fos/CRF/CRF-R1/5-HT in brown or dark brown, weakly expressed evenundetected in NC group, expressed mainly in M group.1.1The effects of JPHSG on c-fos expression in central of D-IBSModel RatCompared with NC group, the expression of c-fos in spinal dorsalhorn,prefrontal cortex, hippocampus and hypothalamus of M group3.1 significantly increased(P<0.01); Compared with M group, all partssignificantly decreased in JPHSG-M and JPHSG-H groups(P<0.01),prefrontal cortex and hippocampus significantly decreased inJPHSG-L(P<0.05,P<0.01), but no significantly different in spinal dorsal hornand hypothalamus with M group(P>0.05); Compared with PC group, all partssignificantly decreased in JPHSG-M and JPHSG-H groups(P<0.05,P<0.01).1.2The effects of JPHSG on CRF expression in central of D-IBSModel RatCompared with NC group, the expression of CRF in prefrontal cortex,hippocampus and hypothalamus of M group significantly increased(P<0.01);Compared with M group, all parts significantly decreased in JPHSG-M andJPHSG-H groups(P<0.01), hippocampus significantly decreased in JPHSG-Lgroup(P<0.01), but no significantly different in prefrontal cortex andhypothalamus with M group(P>0.05); Compared with PC group, all partssignificantly decreased in JPHSG-M and JPHSG-H groups(P<0.05,P<0.01).1.3The effects of JPHSG on CRF-R1expression in central of D-IBSModel RatCompared with NC group, the expression of CRF-R1in prefrontalcortex, hippocampus and hypothalamus of M group significantlyincreased(P<0.01); Compared with M group, all parts significantly decreasedin JPHSG-M and JPHSG-H groups(P<0.01), JPHSG-L group showed nosignificantly different in prefrontal cortex and hypothalamus with Mgroup(P>0.05); Compared with PC group,all parts significantly decreased inJPHSG-M and JPHSG-H groups(P<0.05,P<0.01).1.4The effects of JPHSG on5-HT expression in central of D-IBSModel RatCompared with NC group, the expression of5-HT in prefrontal cortex,hippocampus and hypothalamus of M group significantly increased(P<0.01);Compared with M group, all parts significantly decreased in JPHSG-M andJPHSG-H groups(P<0.01), JPHSG-L group showed no significantly differentin prefrontal cortex and hypothalamus with M group(P>0.05); Compared with PC group, all parts significantly decreased in JPHSG-M and JPHSG-Hgroups(P<0.05,P<0.01).2The effects of JPHSG on c-fos and5-HT content in central of D-IBSModel Rat2.1The effects of JPHSG on c-fos content in central of D-IBS ModelRatCompared with NC group, the content of c-fos in spinal horn andhippocampus of M group significantly increased(P<0.01); Compared with Mgroup, the spinal horn and hippocampus both significantly decreased inJPHSG-M and JPHSG-H groups(P<0.05,P<0.01); Compared with PC group,JPHSG-H group showed significantly decreased (P<0.05), JPHSG-M groupshowed no significantly different with PC group(P>0.05).2.2The effects of JPHSG on5-HT content in central of D-IBS ModelRatCompared with NC group, the content of5-HT in hippocampus of Mgroup significantly increased(P<0.01); Compared with M group, JPHSG-Mand JPHSG-H group decreased significantly(P<0.05,P<0.01); Compared withPC group, JPHSG-H group decreased significantly(P<0.05), JPHSG-L groupshowed no significantly different with PC group(P>0.05).3The effects of JPHSG on CRF and CRF-R1mRNA expression in centralof D-IBS Model Rat3.1The effects of JPHSG on CRF mRNA expression in central ofD-IBS Model RatCompared with NC group, the expression of CRF mRNA in prefrontalcortex and hippocampus of M group significantly increased(P<0.01);Compared with M group, each treatment group decreasedsignificantly(P<0.05,P<0.01); Compared with PC group, JPHSG-M andJPHSG-H groups decreased significantly(P<0.05,P<0.01), PHSG-L groupshowed no significantly different with PC group(P>0.05).3.2The effects of JPHSG on CRF-R1mRNA expression in central ofD-IBS Model Rat Compared with NC group, the expression of CRF-R1mRNA in prefrontalcortex and hippocampus of M group significantly increased(P<0.01);Compared with M group, JPHSG-M and JPHSG-H groups decreasedsignificantly(P<0.05,P<0.01); Compared with PC group, JPHSG-M andJPHSG-H groups decreased significantly(P<0.05,P<0.01), PHSG-L groupshowed no significantly different with PC(P>0.05).Conclusions:The underlying mechanism of JPHSG might be related to deceasedexpression of c-fos/CRF/CRF-R1/5-HT in central. |
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