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目的:观察功能性腹泻脾虚证模型大鼠结肠组织生长激素促释放激素受体(GHSR)表达的变化,探讨发病机制。方法:高乳糖饲料喂养及小平台站立制备功能性腹泻脾虚证大鼠模型,观察大鼠的一般状态变化及粪便情况。免疫组织化学法,蛋白质印迹(Western blotting)和荧光定量PCR(Real-time PCR)检测大鼠结肠组织生长激素促释放激素受体的表达情况。结果:功能性腹泻脾虚的模型大鼠出现精神乏力、消瘦及便溏等现象;免疫组织化学结果显示模型组生长激素促释放激素受体的蛋白表达较正常组表达降低;蛋白质印迹结果显示模型组生长激素促释放激素受体的蛋白表达较正常组表达有差异且有统计学意义(P<0.01);荧光定量PCR结果显示模型组生长激素促释放激素受体mRNA的表达较正常组表达有差异且有统计学意义(P<0.01)。结论:成功构建功能性腹泻脾虚证大鼠模型。功能性腹泻脾虚证大鼠结肠组织生长激素促释放激素受体表达下降。推断生长激素促释放激素受体可能与功能性腹泻脾虚证有相关性。
Objective: To observe the changes of ghrelin (GH) -releasing hormone receptor (GHSR) expression in the colon of rats with functional diarrhea and spleen deficiency syndrome and to explore the pathogenesis. Methods: High-lactose feedstuffs and small platform were set up to prepare functional diarrhea spleen-deficiency syndrome rats model to observe the general state changes and stool conditions. Immunohistochemistry, Western blotting and Real-time PCR were used to detect the expression of growth hormone receptor-releasing hormone receptor in rat colon tissues. Results: The model rats with functional diarrhea and spleen deficiency developed mental retardation, weight loss and loose stools. The results of immunohistochemistry showed that the expression of growth hormone releasing hormone receptor in the model group was lower than that in the normal group. Western blotting showed that model group The expression of somatostatin and releasing hormone receptor was significantly higher than that in normal group (P <0.01). The results of real-time quantitative PCR showed that the expression of growth hormone releasing-releasing hormone receptor mRNA in model group was significantly lower than that in normal group And there was statistical significance (P <0.01). Conclusion: The rat model of functional diarrhea and spleen deficiency was successfully constructed. Expression of growth hormone and releasing hormone receptor in colonic tissue of rats with functional diarrhea and spleen deficiency. Inferred that growth hormone releasing hormone receptor may be associated with functional diarrhea spleen deficiency syndrome.