【作者】 张永东；

【导师】 许剑琴； 穆祥；

【作者基本信息】 中国农业大学 ， 临床兽医学， 2005， 硕士

【摘要】 四君子汤是治疗脾虚证的经典方剂,具有广泛的临床应用价值,目前关于其健脾助消化机制的研究主要集中在对胃肠运动和免疫功能的调节方面,而关于胰腺外分泌与脾虚的相关性研究不多。研究表明胰腺外分泌功能失调是脾虚证的重要病理机制之一,而胆囊收缩素(cholecystokinin,CCK)和血管活性肠肽(vasoactive intestinal peptide,VIP)在胰腺外分泌功能的调节中发挥着非常重要的作用。本试验利用半定量RT-PCR法,以北京鸭肠组织CCK和VIP表达为指标,从调节胰腺外分泌功能方面探讨了四君子汤健脾助消化机制。 1.根据GenBank中鸡的CCK,VIP及内参照鸭的β-激动蛋白(β-actin)的cDNA序列设计CCK,VIP及β-actin的引物,RT-PCR均得到预期大小的片段;PCR产物经纯化、克隆及测序后,同源性分别为90%、94%、100%。 2.用利血平法建立北京鸭实验性脾虚模型,用半定量RT-PCR法检测了十二指肠、空肠CCK和VIP表达的变化。结果,实验性脾虚鸭十二指肠、空肠CCK,VIP mRNA表达均显著低于正常对照组(P<0.01),提示CCK和VIP在脾虚中发挥了重要的作用。 3.四君子汤预防组十二指肠CCK,VIP mRNA表达极显著高于脾虚组(P<0.01),空肠CCK,VIP mRNA显著高于脾虚组(P<0.05),四君子汤预防组和正常对照组之间无显著差异,提示四君子汤对北京鸭实验性脾虚时CCK,VIP mRNA表达降低有明显的抑制作用。 4.自然恢复组十二指肠、空肠CCK,VIP mRNA表达均显著低于正常对照组(P<0.01),治疗Ⅰ、Ⅱ组和正常对照组之间无显著差异。四君子汤治疗Ⅰ、Ⅱ组十二指肠CCK,VIP mRNA表达极显著高于自然恢复组(P<0.01),空肠CCK,VIP mRNA表达显著高于自然恢复组(P<0.05)。治疗Ⅰ、Ⅱ组之间无显著差异。四君子汤对实验性脾虚北京鸭十二指肠、空肠CCK和VIP表达降低有明显的上调作用,提示四君子汤上调CCK和VIP的表达可能是其健脾助消化机理之一。更多还原

【Abstract】 Four Gentlemen decoction is extensively used in clinical practice for the treatment of Spleen deficiency pattern (SDP). However, only a little bit was known about the mechanism of action. Studies have shown that abnormality of exocrine pancreatic secretion was a major pathological mechanism of SDP. Cholecystokinin (CCK) and vasoactive intestinal peptide (VIP) was very potent to stimulate exocrine pancreatic secretion. The aim of the present study is to investigate the effect of Four Gentlemen decoction on the expression of CCK and VIP in duodenum and jejunum of duck SDP models using semiquantitative reverse transcription polymerase chain reaction (RT-PCR).1. Primers were designed according to the sequences of chicken CCK, VIP and duck β -actin cDNA published in the GenBank. The expected sizes of DNA fragments were amplified using RT-PCR technique, the PCR products were cloned into pGEM-T Easy vector for sequence analysis, the results showed that the homologies of sequences were 90%, 94% and 100% respectively.2. The models of SDP were established by injecting Reserpine, expression of CCK, VIP was determined by semiquantitative RT-PCR. The expression levels of CCK, VIP mRNA in experimental SDP were significantly reduced than that in controls (P<0.01). This suggests that CCK. VIP play a key role in SDP.3. No significant differences in the expression levels of CCK, VfP mRNA were found in duodenum and jejunum between Four Gentlemen decoction preventive group and control group. The expression levels of CCK, VIP mRNA in duodenum of SDP group were significantly lower than that of preventive group (P<0.01). The expression levels of CCK, VIP mRNA in jejunum of SDP group were significantly lower than that of preventive group (P<0.05). These results indicate that Four Gentlemen decoction had preventive action on SDP.4. The expression levels of CCK, VIP mRNA in duodenum and jejunum of spontaneous recovery group were significantly lower than that of control group (p<0.01). There were no significant differences in the expression levels of CCK, VIP mRNA between group I, II treated by Four Gentlemen decoction and control group. In duodenum, the expression levels of CCK, VIP mRNA in group I, II treated by Four Gentlemen decoction were significantly higher than that of spontaneous recovery group (P<0.01), in jejunum, that is P<0.05. No significant differences were found between group I and group II. These findings suggest that Four Gentlemen decoction may exert their beneficial therapeutic action on SDP, at least in part, by upregulated CCK, VIP mRNA levels in duodenum and jejunum.更多还原