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目的:探讨四神丸对脾肾阳虚型泄泻模型大鼠下丘脑、十二指肠组织中胃促生长素(Ghrelin)及其受体(GHSR)蛋白和mRNA表达的影响。方法:将72只SPF级SD大鼠随机分为正常组、模型组、得舒特组、四神丸高、中、低剂量组,每组12只。除正常组外,其余各组均采用灌服腺嘌呤和番泻叶水煎剂复制脾肾阳虚型泄泻大鼠模型。连续给药14d后观察各组大鼠一般状态,ELISA法检测十二指肠组织中Ghrelin含量,Western Blot、RTPCR技术检测下丘脑、十二指肠中Ghrelin、GHSR蛋白和mRNA表达情况。结果:模型组大鼠一般状态虽有所改善但腹泻仍持续存在,与正常组比较,十二指肠中Ghrelin含量及下丘脑、十二指肠中Ghrelin、GHSR蛋白和mRNA表达均明显增高(P<0.01)。与模型组比较,四神丸高、中剂量组和得舒特组十二指肠Ghrelin含量降低,下丘脑、十二指肠中的Ghrelin、GHSR mRNA表达下降(P<0.05,P<0.01),四神丸高、中、低剂量组和得舒特组均可下调下丘脑、十二指肠中的Ghrelin、GHSR蛋白表达(P<0.05,P<0.01),且以四神丸高剂量组尤为明显。四神丸中剂量组脑肠中Ghrelin、GHSR的表达与得舒特组比较均无显著性差异。结论:四神丸对脾肾阳虚泄泻模型大鼠的作用机制可能与下调Ghrelin、GHSR的蛋白和基因过表达有关。
Objective: To investigate the effect of Si Shen Pill on the protein and mRNA expression of ghrelin and its receptor (GHSR) in hypothalamus and duodenum of rats with spleen - yang deficiency type diarrhea model. Methods: Seventy-two Sprague-Dawley rats were randomly divided into normal group, model group, dextran group and Sishenwan high, medium and low dose groups, 12 rats in each group. In addition to the normal group, the other groups were fed with adenine and senna decoction of spleen and kidney yang deficiency diarrhea model rats. After administration for 14 days, the general state of rats in each group was observed. The content of Ghrelin in duodenum was detected by ELISA. The expression of Ghrelin and GHSR protein in hypothalamus and duodenum was detected by Western Blot and RTPCR. Results: Although the general condition of the rats in the model group was improved, diarrhea persisted. Compared with the normal group, the Ghrelin content in the duodenum and the expression of Ghrelin and GHSR protein and mRNA in the hypothalamus and duodenum were significantly increased P <0.01). Compared with the model group, the content of Ghrelin in duodenum of Sishenmao, middle-dose and dextro-treated groups decreased, the expression of Ghrelin and GHSR mRNA in hypothalamus and duodenum decreased (P <0.05, P <0.01) (P <0.05, P <0.01). The expressions of Ghrelin and GHSR in hypothalamus and duodenum were significantly decreased in high, medium and low dose groups and DST group Group is particularly obvious. The expression of Ghrelin and GHSR in the middle intestine of Sishen pill group was not significantly different from that of dextrose group. Conclusion: The mechanism of Sishen pill on the rats with diarrhea due to spleen and kidney yang deficiency may be related to the down-regulation of Ghrelin and GHSR protein and gene overexpression.