阳虚便秘动物模型的建立及济川煎治疗作用研究

发布时间：2018-01-12 01:35

本文关键词：阳虚便秘动物模型的建立及济川煎治疗作用研究　出处：《山西省中医药研究院》2017年硕士论文　论文类型：学位论文

【摘要】：目的建立大鼠阳虚便秘动物模型,并在此基础上对济川煎治疗阳虚便秘的作用及作用机制进行初步的探讨研究。方法关于阳虚便秘病证结合动物模型建立部分:用一组大鼠按照文献方法即食用白醋加活性炭冰水作为造模因素来制作大鼠阳虚便秘动物模型,用另外一组大鼠按照改进文献方法以后即4℃食用白醋加活性炭冰水再加上止泻药物复方地芬诺酯的办法建立大鼠阳虚便秘动物模型,并设置空白对照组。然后分别在造模因素停止后第1天和去除造模因素后第7天对各组大鼠同时进行检测评价分析,具体检测从以下几方面进行:观察实验大鼠在模型建立过程中的一般状态并进行评分记录,观察实验大鼠首粒排便时间和排便粒数,用生物机能测定仪检测实验大鼠的结肠肌电改变,用酶联免疫吸附法对实验大鼠体内乙酰胆碱酯酶(Ach E)、胃动素(MTL)、生长抑素(SS)、环磷酸腺苷(c AMP)、环磷酸鸟苷(c GMP)、蛋白激酶A(PKA)、蛋白激酶C(PKC)的含量变化进行检测。最后对所得数据用SPSS16.0进行统计分析得出结论。有关济川煎治疗阳虚便秘作用及其机制初步研究部分:复制上述改进文献方法后的阳虚便秘动物模型,然后对其进行随机分组,分别为模型组、阳性药物对照组、济川煎高(8.63g/kg)、中(4.31g/kg)、低(2.16g/kg)剂量组,加之空白对照组共计6组。阳性药物对照组大鼠使用莫沙必利进行治疗,济川煎高、中、低剂量组大鼠分别使用济川煎的相应剂量进行灌胃治疗,空白对照组和模型组大鼠在此期间给予同等剂量的蒸馏水灌胃。在治疗给药结束后对各组大鼠同时进行检测评价分析,具体检测项目同上,最后所得数据用SPSS16.0进行统计分析得出结论。结果关于动物病证结合模型建立部分:在造模因素停止后第1天,两模型组大鼠的一般状态都较差,均出现了腹胀、体重减轻、皮毛枯槁乍立、倦怠蜷缩、反应迟缓、粪便干硬等征象,与空白对照组相比也都出现了首粒排便时间后延、排便粒数减少,结肠肌电慢波频率减慢、振幅增大的现象(P㩳0.05或P㩳0.01),也都表现出血浆MTL含量下降,SS水平升高,血清Ach E含量下降,c AMP/c GMP比值降低,下丘脑组织上清中PKA水平下降,PKC水平升高的变化(P㩳0.05或P㩳0.01)。但改进文献方法后的模型组(模型Ⅱ组)大鼠病理状态与文献方法模型组大鼠(模型Ⅰ组)相比其表现则更为明显。去除造模因素7天后,模型Ⅰ组大鼠基本自愈,精神状态、活动度、排便状况、结肠运动情况,Ach E、MTL、SS、c AMP/c GMP、PKA、PKC的变化等与空白对照组大鼠相比几乎没有差别,虽然没有恢复至与空白组大鼠同等水平,但其数值与空白组大鼠相应数值比较,其差异已经没有统计学意义了。而模型Ⅱ组大鼠病理状态依然明显,各项病理检测指标与空白对照组大鼠相比其差异依然具有统计学意义(P㩳0.05或P㩳0.01)。关于济川煎治疗阳虚便秘作用及其机制初步探讨部分:济川煎高、中剂量组大鼠一般状态恢复较好,相比较于模型组大鼠其体重也有所增长,同时首粒排便时间缩短,排便粒数增加,结肠肌电慢波频率加快、振幅下降(P㩳0.05或P㩳0.01),血浆MTL含量升高,SS水平下降,血清Ach E含量升高,c AMP/c GMP比值上升,下丘脑组织上清中,PKC水平降低,与模型组相比有显著差异(P㩳0.05或P㩳0.01)。济川煎低剂量组大鼠的状态与模型组大鼠相比有所改善,但没有显著差异。阳性药物对照组大鼠状态的改善则仅限于便秘相关方面,如与模型组大鼠相比首粒排便时间缩短、排便粒数增加,结肠肌电慢波频率升高,振幅降低,血浆MTL含量升高,SS水平下降(P㩳0.05或P㩳0.01)等,而其皮毛色泽、精神状态、体重的改变则不大,c AMP/c GMP比值、PKC水平的变化与模型组大鼠相比则没有差异。结论4℃白醋、活性炭冰水加复方地芬诺酯灌胃可建立起比较稳定的阳虚便秘动物模型,模型动物的病理特征与阳虚便秘的临床症状相类似,并存在肠神经递质和胃肠激素分泌混乱。济川煎高、中剂量组能不同程度地改善阳虚便秘模型大鼠的各项症状指征,其作用机制考虑可能与其影响环核苷酸、蛋白激酶、肠神经递质和胃肠激素变化有关。
[Abstract]:Objective to establish a rat animal model of Yang deficiency and constipation, were preliminary studied the effect and mechanism of action and on the basis of Jekawa in the treatment of Yang deficiency and constipation. Methods of Yang deficiency and constipation disease combined with syndrome animal model: one group of rats according to the method of instant white vinegar and activated carbon as the ice model to produce a large the rat animal model of Yang deficiency and constipation, with another group of rats after the improved method is in accordance with the literature 4 C vinegar plus charcoal ice plus antidiarrheal drug compound diphenoxylate to establish rat animal model of Yang deficiency and constipation, and set the blank control group. Then the rats in the first days after the removal of stop factors and model after seventh days of rats in each group were detected at the same time analysis, specific detection from the following aspects: the rats were observed in the model building process and the general state The score records, observed the first defecation time and defecation grain grain number, change the colonic electromyography measurement instrument detection of biological function of experimental rats, rats of acetylcholinesterase in vivo by enzyme-linked immunosorbent assay (Ach E), motilin (MTL), somatostatin (SS), cyclic adenosine monophosphate (C AMP), cyclic guanosine monophosphate (C GMP), protein kinase A (PKA), protein kinase C (PKC) contents were detected. Finally the data were analyzed by SPSS16.0. The conclusion part Jichuanjian treatment effect and mechanism of Yang deficiency and constipation: a preliminary study on animal model of Yang deficiency and constipation of the improved method of literature after then, were randomly assigned to it, respectively, model group, positive drug group, Jekawa Ta (8.63g/kg), in (4.31g/kg) (2.16g/kg), low dose group and blank control group, a total of 6 groups. The positive drug control group rats using mosapride in For treatment, the corresponding Jichuanjian high dose and low dose groups of rats were used Jichuanjian by intragastric administration of distilled water during the treatment, the control group and model group rats were then given the same dose of intragastric administration in the treatment. After the end of the rats were detected at the same time analysis, specific test items above, finally, the data statistical analysis using SPSS16.0 model. The results of part of the combination of disease and syndrome animal model: in the stop after first days, the general state of the two rats in the model group were poor, there were abdominal distension, weight loss, haggard fur first, burnout curled up, slow response, dry hard stool signs, compared with the control group also appeared in the first time of defecate after the extension, the number of defecate decreases, colonic electromyography decrease of slow wave frequency, amplitude increased (P? 0.05 or P? 0.01), also showed a decrease in plasma MTL content SS, elevated levels of serum Ach, decrease the content of E, C, AMP/c and GMP ratio decreased, the level of PKA in hypothalamus was decreased, PKC levels increased (P? 0.05 or P? 0.01). But the improvement method of literature model group (model II Group) rats pathological state and method of literature model group rats (model group) compared to its performance is more obvious. The removal model for 7 days, model group rats basic self-healing, mental state, activity, defecation condition, colonic motility, Ach, E, MTL, SS, C AMP/c GMP, PKA, PKC change compared to almost no difference with the rats in control group, although not recovered with the blank group the same level, but the value of the rats in the blank group the corresponding numerical comparison, the difference was not statistically significant. The pathological state of rat model of group II is still obvious, the pathological detection index and the rats in control group compared the difference in However, with statistical significance (P? 0.05 or P? 0.01). And to explore the mechanism of treatment of constipation: part Jichuanjian Yang Jichuanjian high dose group, the general condition of rats recovered well, compared with the model rats, the body weight also increased, while the first defecate time, defecate number increase, colon myoelectric slow wave frequency speed, the amplitude decreased (P? 0.05 or P? 0.01), plasma MTL increased and SS level decreased, serum Ach content of E, C AMP/c GMP ratio was increased, the hypothalamus in the supernatant of PKC decreased, there was significant difference compared with the model group (P? 0.05 or P? 0.01 Jichuanjian). Low dose group rats status improved compared with the model rats, but no significant difference. To improve the positive drug control group in rats is limited to constipation related aspects, such as the first grain of defecation was shortened compared with the rats in the model group, the number of defecate increase, Colonic myoelectric slow wave frequency increases, the amplitude decreased, plasma MTL increased and SS decreased (P? 0.05 or P? 0.01), but their fur color, mental state, weight change is small, the ratio of GMP C AMP/c, PKC level and the changes of rats in the model group compared with no difference. Conclusion 4 C white vinegar, activated carbon and water compound diphenoxylate gavage can establish stable animal model of Yang deficiency and constipation, clinical symptoms and pathological features of Yang deficiency and constipation animal models are similar, and the existence of intestinal neurotransmitters and gastrointestinal hormone secretion disorder. Jichuanjian high dose group could improve the symptoms of Yang deficiency constipation model rat indications, considering its mechanism of action may be related to its effect of cyclic nucleotide, protein kinase, intestinal neurotransmitters and gastrointestinal hormones.

【学位授予单位】：山西省中医药研究院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5;R-332

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