运脾开胃法对VIP通路中PKA及NO调节机制的研究

发布时间：2018-10-29 08:22

【摘要】：目的：从VIP受体及其下游信号因子的调节机制来探讨运脾开胃法治疗厌食症的深层机理,进而有效地指导临床。方法：选用运脾开胃复方作为运脾开胃治法的工具药,用空白、低、中、高剂量含药血清作用于人体胃窦平滑肌细胞,其中低剂量等同于人体等效剂量。运用免疫印迹法,从蛋白水平测定VIP受体(VPAC2)及其下游信号因子eNOS、p-eNOS、PKA的表达；同时,运用硝酸还原酶法测定细胞内NO浓度变化趋势,分析各组数据及差异的内在涵义。结果：生理状态下,人胃窦平滑肌细胞只有在经中、高剂量含药血清作用后,VPAC2、eNOS、 peNOS蛋白表达水平降低(P0.05),但中高剂量组之间无明显差异;PKA蛋白水平表达在各组之间皆无统计学意义。除空白组外经含药血清作用后的人胃窦平滑肌细胞内N0浓度皆有降低(P0.05),但低、中、高组之间并无明显差别。结论：运脾开胃工具药在细胞实验中对胃肠肽的信号传导调节是级联反应,VIPR与下游信号分子表达量的变化趋势是一致的,在达到人体5倍等效剂量之后才发挥其药理作用。然而,产生的药理学效应并不随剂量的增加而增强。运脾开胃复方对VIP的调节可通过NO相关途径来实现,与cAMP依赖的蛋白激酶途径无明显关联。
[Abstract]:Objective: to explore the underlying mechanism of VIP receptor and its downstream signaling factors in the treatment of anorexia. Methods: the compound of spleen and appetizer was chosen as the tool drug for the treatment of spleen and stomach. The serum containing drugs in blank, low, middle and high doses acted on human gastric antral smooth muscle cells, in which the low dose was equivalent to the human equivalent dose. The expression of VIP receptor (VPAC2) and its downstream signal factor (eNOS,p-eNOS,PKA) were detected by Western blot. At the same time, the change trend of intracellular NO concentration was measured by nitrate reductase method, and the inner meaning of each group's data and difference was analyzed. Results: under physiological condition, the expression of VPAC2,eNOS, peNOS protein in human gastric antral smooth muscle cells decreased only after the action of medium and high dose serum (P0.05), but there was no significant difference between the middle and high dose groups. There was no significant difference in the expression of PKA protein between each group. In addition, the concentration of N0 in human gastric antral smooth muscle cells was decreased (P0.05), but there was no significant difference between the low, middle and high groups. Conclusion: the regulation of gastrointestinal peptide signal transduction by spleen appetizer is a cascade reaction, and the change trend of VIPR and downstream signal molecule expression is the same, and the pharmacological effect is only shown after reaching the equivalent dose of 5 times in human body. However, the pharmacological effect does not increase with the increase of dose. The regulation of YFQF on VIP may be achieved by NO related pathway, but not by cAMP dependent protein kinase pathway.
【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R259

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