【摘要】：目的：從VIP受體及其下游信號(hào)因子的調(diào)節(jié)機(jī)制來探討運(yùn)脾開胃法治療厭食癥的深層機(jī)理,進(jìn)而有效地指導(dǎo)臨床。方法：選用運(yùn)脾開胃復(fù)方作為運(yùn)脾開胃治法的工具藥,用空白、低、中、高劑量含藥血清作用于人體胃竇平滑肌細(xì)胞,其中低劑量等同于人體等效劑量。運(yùn)用免疫印跡法,從蛋白水平測(cè)定VIP受體(VPAC2)及其下游信號(hào)因子eNOS、p-eNOS、PKA的表達(dá)；同時(shí),運(yùn)用硝酸還原酶法測(cè)定細(xì)胞內(nèi)NO濃度變化趨勢(shì),分析各組數(shù)據(jù)及差異的內(nèi)在涵義。結(jié)果：生理狀態(tài)下,人胃竇平滑肌細(xì)胞只有在經(jīng)中、高劑量含藥血清作用后,VPAC2、eNOS、 peNOS蛋白表達(dá)水平降低(P0.05),但中高劑量組之間無明顯差異;PKA蛋白水平表達(dá)在各組之間皆無統(tǒng)計(jì)學(xué)意義。除空白組外經(jīng)含藥血清作用后的人胃竇平滑肌細(xì)胞內(nèi)N0濃度皆有降低(P0.05),但低、中、高組之間并無明顯差別。結(jié)論：運(yùn)脾開胃工具藥在細(xì)胞實(shí)驗(yàn)中對(duì)胃腸肽的信號(hào)傳導(dǎo)調(diào)節(jié)是級(jí)聯(lián)反應(yīng),VIPR與下游信號(hào)分子表達(dá)量的變化趨勢(shì)是一致的,在達(dá)到人體5倍等效劑量之后才發(fā)揮其藥理作用。然而,產(chǎn)生的藥理學(xué)效應(yīng)并不隨劑量的增加而增強(qiáng)。運(yùn)脾開胃復(fù)方對(duì)VIP的調(diào)節(jié)可通過NO相關(guān)途徑來實(shí)現(xiàn),與cAMP依賴的蛋白激酶途徑無明顯關(guān)聯(lián)。
[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.
【學(xué)位授予單位】：南京中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R259

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