| Objective:Diarrhea-predominant irritable bowel syndrome (IBS-D) is a common clinical disorders of gastrointestinal function. It has unique advantages in treatment based on syndrome differentiation of traditional Chinese medicine. Liver-depression and Spleen-deficiency syndrome is a common pattern in TCM. Tong- xie-yao-fang is the widely method used to treating this disease in clinical practice.At present, IBS-D has lack of recognized experimental study on animal model of syndrome in TCM. The primary prerequisite for treatment this disease is to established successfully model of Liver-depression and spleen-deficiency syndrome type of traditional Chinese medicine.Although it has a large number of clinical studies have shown that pain treatment IBS-D effective for treating diarrhea,but lack of experimental study on traditional Chinese medicine theory. By studying comparatively,we want to find the experimental supports for Preparation of this study as a model of TCM syndrome and Tong-xie-yao-fang treatment IBS-D high-efficiency.Further on improve the theoretical study of irritable bowel syndrome and provide theoretical and experimental support for clinical treatment of selected drugs.Methods:Choose healthy SPF 120 SD rats(half male and female),8 weeks old, weighing 190±10g. Adaptability of feeding a week later,12 randomly selected (male half) as a normal group (N); the remaining rats started to modeling.Combining Hung Lu, Deirdre described modeling methods such as using chronic unpredictable mild stress combined with high fat diet high sugar, high heat and high temperature,high humidity environment to establish close clinical symptoms in patients with Liver depression and spleen deficiency of TCM rat model of IBS-D. The success criteria of the identification model include:Status and weight gain, rats choosing dejecta character score, and intestinal pain threshold sensitivity, open box scores and preference in syrup. According to above four items integrated points sort, selection meet standard of big rat. According to random to diyided into model group (M),Tong-xie-yao-fang 7 days treatment group (T7),Live Bifidobacterium Preparation-Oral 7 days treatment group (B7), and Tong-xie-yao-fang14days treatment group (T14),Live-Bifidobacterium Preparation-Oral 14days treatment group (B14).Each group have 12 rats (male and female each half).Drugs:N and M group was given the same volume of normal saline for gavage experiment in rats has daily 14 consecutive days;T7groupwasgiven Tong-xie-yao-fang decoction (7.5g/kg.d-1) of prednisone for 7 consecutive days; B7 group given daily doses of Bifidobacterium (2×107 CFU/ml.d-1) of prednisone for 7 consecutive days; T14 group was given Tong-xie-yao-fang decoction (7.5g/kg.d-1) of prednisone for gavage for 14 consecutive days; B14group given daily doses of Bifidobacterium suspensions (2×107 CFU/ml.d-1) of prednisone for 14 consecutive days.Sample based and the processing:And after treatment, After heart blood preservation, the rats serum was tested with Alliance immune method about the contents of 5-HT and VIP. Experimental data obtained are used in statistical analysis of SPSS 17.0 in the process, satisfy normality of the distribution of the measurement data, data obtained by mean standard deviation (x±s); among groups using one-way analysis of variance, and homogeneity of variance between groups by using LSD, variances are not aligned groups were compared using the DunnettT3 method.Results:1.General status in rats:rats before the model whose hair smooth, responsive and agile, good spirit, normal stools. Modeling after the end, compared with the normal group, less gloss coat color in the model group, even with vertical hair, reduces movement, loss of appetite, like curling up, congregate, have been slow, stool softening or unfounded, anus or faeces dirt attached. After the end of treatment, compared with normal group, Bifidobacterium tong-xie-yao-fang medicine group and rats General State has improved significantly, appetite enhancement, increased activity, relatively sensitive, loose stools or reduction. 2.Body weight change pattern:(1) Modeling after the end:compared with the normal, slow weight gain in the model group, statistically significant weight gain (p=0.00,p<0.05).(2) 7 days after the end of treatment:Compared with the normal group, model groups there is statistically significant difference between the(p=0.00,p<0.01),Tong-xie yao-fang and Bifidobacterium group no statistical difference(p=0.04,0.02,p<0.05); Compared with the Group of tong-xie-yao-fang, Bifidobacterium Group had no statistically significant differences(p>0.05)。(3) 14 days after the end of treatment：compared with the normal group, and model groups there is statistically significant difference between the(p=0.00,p<0.01),Tong-xie-yao-fang group and Bifidobacterium group no statistical difference (p=0.06,0.07,p>0.05);Compared with control group, Tong-xie-yao-fang group and Bifidobacterium group body weight increases rapidly, there was a significant difference in(p<0.05);Compared with the Group of tong-xie-yao-fang, Bifidobacterium Group had no statistically significant differences(p>0.05).3.Rat model of evaluation results:(1) Rat body weight changes during model:model 14 days later, model group slow weight gain. Compared with the control group, group weight there is statistically significant difference(p=0.02,p<0.05);Modeling the 28th day, compared with the control group, weight of each model group had statistically significant difference between the (p=0.00,p<0.01).(2) Rat droppings trait scoring change:modeling after the dejecta character of each model Group score was significantly higher than the control group (p=0.00);Compared with the control group, each model group differences were significant statistically(p<0.01).(3) Changes of intestinal sensitivity and pain threshold in rats:model after the end of each model group reduced pain threshold in rats, increased intestinal sensitivity (p=0.00);Compared with the control group, there was a significant difference statistically significant(p<0.01).(4) Open box test score results:modeling after the Open box of rats in each group exercise scores lower (p=0.00);Compared with the control group, there was a significant difference statistically significant(p<0.01).(5) Sugar preference changes:model after the sugar intake in each model group decreased (p=0.00);Compared with the control group, sugar preference there was significant statistical difference(p<0.01).4.Changes of serum 5-HT and VIP of rats:(1) 7 days after the end of treatment, rats in each group of changes of serum 5-HT, VIP:normal group minimum, models the highest, tong-xie-yao-fang and Bifidobacterium group and between groups.(2)Treatment for 14 days after the end of content changes of serum 5-HT and VIP of rats in each group:Tong-xie-yao-fang group and Bifidobacterium group close to normal. Compared with the normal group, model 5-HT, VIP content there is statistically significant difference between the(p<0.01).Conclusion:(1)It can build Liver-depression and spleen deficiency model IBS-D for TCM with chronic unpredictable mild stress combined with high fat diet high sugar, high heat, high temperature and high humidity environment.By tong-xie-yao-fang prescription to treat IBS-D has proved traditional Chinese medicine treatment of Liver-depression and spleen-deficiency syndrome efficiently.(2) Experimental studies have found that in 7 days after treatment, the serum 5-HT and VIP of Bifidobacterium group and tong-xie-yao-fang prescription effect not significant;In the 14th day after treatment, the serum 5-HT and VIP of Bifidobacterium group and tong-xie-yao-fang group be close to the normal group. It confirmed that Tong-xie-yao-fang has effective in treating IBS-D of traditional Chinese medicine. |
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