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Objective:To observe the effect of Changji'an prescription on intestinal permeability in diarrhea-predominant irritable bowel syndrome (IBS-D) rats and explore its mechanism for treatment of IBS-D. Method:Male SD neonatal rats were randomly divided into five groups:normal group, model group, pinaverium bromide group(0.018 g·kg-1), high-dose(33.48 g·kg-1) and low-dose (16.74 g·kg-1)Changji'an prescription groups. Except for the normal group, the IBS-D model was established by the combination of maternal and infant separation+acetic acid stimulation+restraint stress. After drug treatment, the ultrastructure of rat intestinal mucosa was observed by using transmission electron microscopy and the plasma D-lactate level was detected by enzyme-linked immunosorbent assay (ELISA). The expression levels of tight junction proteins Occludin and Claudin-1 were detected by Western blot. The mRNA expression levels of Occludin,Claudin-1 and zonula occluden(ZO)-1 were detected by real time polymerase chain reaction (Real-time PCR). Result:As compared with the normal group, the intestinal mucosal epithelial cells were damaged in IBS-D model group, and the microvilli arrangement was sparse and tight junction was widened, and some were not obvious,and the plasma D-lactate level in IBS-D rats was increased significantly (PPD-lactate level in pinaverium bromide group and high-dose Changji'an prescription group was significantly decreased (PD-lactate level in the low-dose group Changji'an prescription group had a tendency to decrease with no statistical difference. The mRNA and protein expression levels of Occludin and Claudin-1 and the mRNA expression of ZO-1 in the colon of rats in each administration group were higher than those in the model group (PConclusion:The therapeutic effect of Changji'an prescription on IBS-D may be achieved by improving the intestinal permeability.
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OBJECTIVE:To investigate the effect of Changji’an capsules on abdominal pain and the molecular mechanism re-lated to calcitoningene-related peptide(CGRP)and corticosterone(CORT)in diarrhea-predominant irratable bowel syndrome(IBS-D)model rats. METHODS:After the rat models of IBS-D were established by the method of separation of breast mick combined with stimulation with acetic acid,the rats were randomly divided into model group(isometric normal saline),pinaverium bromide group(0.018 g/kg),and Changji’an capsules high,medium and low dose groups(2.812 g/kg,1.406 g/kg and 0.703 g/kg),and another SD rats were included in the normal control group(isometric normal saline). The drugs were given to the rats once a day for consecutive 14 d,ig. Injection of normal saline method was adopted to determine the rat’s sensibility to abdominal pain. The en-zyme-linked immunosorbent assay(ELISA)was adopted to determine the content of CORT in the serum of the rats. Subjected to re-verse transcription polymerse chain reaction(RT-PCR)was adopted to determine the expression of CGRP mRNA in the hypothala-mus and colon tissues of the rats. RESULTS:Compared with normal control group,threshold values of arching the back and stick-ing out the abdomen were decreased,the content of CORT in serum and expression of CGRP mRNA in the hypothalamus and co-lon tissues in model group were increased,with significant difference(P<0.01). Compared with model group,threshold values of arching the back and sticking out the abdomen were increased,the content of CORT in serum and expression of CGRP mRNA in the hypothalamus and colon tissues in pinaverium bromide group and Changji’an capsules high and medium dose groups were de-creased;the threshold value of arching the back in Changji’an capsules low dose group were increased,with significant difference (P<0.01 or P<0.05). CONCLUSIONS:Changji’an capsules can improve the abdominal pain in rats with IBS-D by a mechanism that may be related to the decrease in the expression of CGRP mRNA in the hypothalamus and colon tissues and the reduc-tion of the content of CORT in serum.
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OBJECTIVE:To establish the microemulsion TLC for the separation and identification of various effective compositions in Changji'an prescription.METHODS: The compositions in Changji'an prescription were identified on polyamide film(fixed phase) with five distribution ratios of sodium lauryl sulphate-n-butanol-skellysolve C-H2O microemulsions as developing agents.The effects of factors such as the water content of the microemulsion and the temperature on the separation efficacy were studied.RESULTS: With microemulsion(75% water content) as developing agent,Rhizoma Coptidis in Changji'an prescription was detected;with microemulsion(75% water content)-formic acid(9.0∶1.0) as developing agent,Corydalis yanhusuo,Saposhnikovia divaricata,chlorogenic acid and Radix et Rhizoma Glycyrrhizae in Changji'an prescription were detected.CONCLUSION: Microemulsion TLC is simple,environmental-friendly,sensitive with ideal separation efficacy,and it can be used for the separation and identification of various compositions in Chinese patent medicines.
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Objective To observe the effect of Changji’an Prescription (CP) on depressing the mast cells in the irritable bowel syndrome(IBS)rats. Methods The postnatal rats aged 8～21 days were given continous inflammation stimulation inside the colon to replicate the animal model of the IBS. The number of mast cells and their degranulation were observed by modified Toluidine Blue staining method under light microscope and electron microscope. Results The pressure threshold value of rats raising their abdomen and back in the model group was obviously lower than that in the normal group (P
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Objective To observe the effect of Changji'an Prescription(CP) on T lymphocytes count and 5-hydroxytryptamine(5-HT) secretion in rats with diarrhea predominant irritable bowel syndrome(IBS).Methods Forty-six neonatal SD rats were randomized into normal group,model group,low-,middle-and high-dose CP groups(0.94,1.88 and 3.76g?kg-1?d-1 respectively),and Bupi Yifei Pills group(6.76g?kg-1?d-1).Continuous internal rectal stimulation with acetate was used to induce the rat model of intestinal hypersensitive IBS.Flow cytometer was used for the quantitative analysis of peripheral levels of T lymphocytes subsets CD+3,CD+4 and CD+8,and immunohistochemical method was used to observe 5-HT-positive cells number and the immunostaining intensity in the bowels of rats.Results In the model group,the defecation volume and the intestinal sensitivity were increased,CD+4 level and CD+4/CD+8 decreased,CD+8 level increased,5-HT-positive cells number and the immunostaining intensity elevated(P