| Objetive: To establish the rat model of yang-deficiency and constipation syndrome and to preliminarily study the effect and mechanism of Ferry Brew on the base of the established model.Methods : Part1: The establishment of rats model combined Disease and Zheng.Experimental animals were divided into three groups: modelⅠ,modelⅡ,and the blank control group.The rats of modelⅠwere treated with activated carbon plus edible white vinegar suspension in frozen water according to the literature;the rats of model Ⅱwere treated with 4℃suspension of activated carbon plus edible white vinegar combined compound diphenoxylate,a kind of Antidiarrheal drug,according to a improved method on the foundation of the methods of literature;the rats of the blank control group were treated with distilled water.Then,evaluated and analyzed the rats from three groups via observing the general state of the experimental rats during the model establishment,recorded in marking,the first defecation time and the number of defecation,testing changes in colonic electromyography in rats using biological function analyzer,and measuring acetylcholin esterase(Ach E),motilin(MTL),somatostatin(SS),cyclic adenosine monophosphate(c AMP),guanosine monophosphate(c GMP),protein kinase A(PKA),and protein kinase C(PKC)were measured by enzyme-linked immunosorbent assay A(ELISA),on the first day after the modeling factor was stopped and the 7th day after removing the modeling factor.Finally,the data were analyzed by SPSS16.0.Part 2: the preliminary study of the effect and mechanism of Ferry Brew on the base of the established model.The animal model of yang-deficiency and constipation Syndrome was reconstructed by the improved method based on the methods of literature,and then the rats were randomly divided into 6 groups: model group,positive drug control group,Ferry Brew high(8.63g/kg),medium(4.31g/kg)and low(2.16g/kg)dose group,and control group.The rats in the positive control group were treated with mosapride;the rats in the high,medium and low dose groups were treated with the corresponding dose of Ferry Brew,and the rats in the blank control group and the model group were given the same dose of distilled water.At the end of the treatment,the rats in each group were tested and evaluated at the same time via the above items.The final data were analyzed by SPSS16.0.Results: Part1: The establishment of rats model combined Disease and Zheng.On the first day after the modeling factor was stopped,the general state of the two model groups were poor.The rats of two model groups had weight loss,fatigue,curl up,slow response,dry stool and other signs compared with the blank control group.And the difference was significant(P <0.001).The first defecation time was delayed(P <0.05 or P<0.01),the number of defecation particles decreased(P <0.05 or P <0.01),the frequency of colonic electromyography slowed down and the amplitude increased(P <0.05 or P <0.01),compared with the blank control group.The results showed that the content of plasma MTL decreased,the level of SS was increased,the content of serum Ach E decreased,the ratio of c AMP / c GMP decreased,the level of PKA decreased and the level of PKC increased in the supernatant of the hypothalamus homogenate(P <0.05 or P <0.01),compared with the blank control group.However,the pathological status of the model group Ⅱ(established by the improved method on the foundation of the methods of literature)was significantly higher than that of the model group Ⅰ(established by the method from literature).On the 7th day after removing the modeling factor,the changes of Ach E,MTL,SS,c AMP / c GMP,PKA and PKC in the model group Ⅰ were similar to those in the blank control group,and there was no significant difference between the control group.Although the rats of mode groupⅠdidn’t restored to the same level as the blank group rats,the value compared with the corresponding value of the blank group,wasn’t statistically different.The pathological status of model group Ⅱ was still obvious,and the difference of the pathological indexes between the model group Ⅱ and the blank control group was statistically significant(P <0.05 or P <0.01).Part 2: the preliminary study of the effect and mechanism of Ferry Brew on the base of the established model.The weight of the rats in the high dose group and the middle dose group were better than those in the model group.And the first defecation time was shortened,the number of defecation increased,the frequency of colonic myoelectric quickened and amplitude decreased(P<0.05 or P <0.01),plasma MTL content increased,SS level decreased,serum Ach E content increased,c AMP / c GMP ratio increased,PKC levelsin the supernatant of the hypothalamus homogenate decreased compared with the model group(P <0.05 or P <0.01).The state of the rats in the low dose group was improved compared with the model group,but there was no significant difference.The improvement of the state of the positive control group was limited to the constipation-related aspects.Compared with the model group,the first defecation time was shortened,the number of defecation increased,the frequency of colonic muscle slow wave increased,the amplitude decreased,but the plasma MTL content and the change of skin color,mental state and body weight were not significant.The changes of c AMP / c GMP ratio and PKC level were not different compared with those of model group.Conclusion: Using 4 ℃ suspension of white vinegar,activated carbon ice water combined with compound diphenoxylate could establish relatively stable yang-deficiency and constipation syndrome Animal model.Model animal pathology was similar to clinical symptoms of yang deficiency constipation.And there were intestinal neurotransmitter and gastrointestinal hormone secretion disorder.Ferry Brew high and medium doses could improve the symptoms of Yang deficiency constipation model rats.And the mechanism of the efficacy might be related to the changes of cyclic nucleotide,protein kinase,intestinal neurotransmitter and gastrointestinal hormone secretion. |
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