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目的探讨四神丸对脾肾阳虚泄泻模型大鼠十二指肠组织三磷酸肌醇(IP3)、钙调素(Ca M)、肌球蛋白轻链激酶(MLCK)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,采用ELISA法检测十二指肠组织中IP3的含量,Western Blot、免疫组织化学法分别检测十二指肠中Ca M和MLCK的蛋白表达。结果模型组大鼠十二指肠组织匀浆中IP3含量及十二指肠中Ca M、MLCK蛋白表达均明显增高,与正常组比较差异有显著性(P<0.01)。与模型组比较,四神丸高、中剂量组和得舒特组IP3含量降低(P<0.05或P<0.01),四神丸各剂量组和得舒特组Ca M、MLCK蛋白表达下降(P<0.05或P<0.01);且四神丸高剂量组优于低剂量组(P<0.05),而四神丸中剂量组上述指标改变与得舒特组比较差异不显著(P>0.05)。结论四神丸对脾肾阳虚泄泻模型大鼠的作用机制可能与调节十二指肠IP3-Ca M-MLCK信号途径指标有关。
Objective To investigate the effect of Si Shen Pill on the expression of inositol 1,4,5 - trisphosphate (IP3), calmodulin (Ca M) and myosin light chain kinase (MLCK) in the duodenum of rats with spleen - yang - Yang diarrhea model. Methods Seventy-two Sprague-Dawley (SD) SD rats were randomly divided into normal group, model group, dextran group and Sishen Pill 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 content of IP3 in duodenum was detected by ELISA. The protein expressions of Ca M and MLCK in duodenum were detected by Western Blot and immunohistochemistry respectively. Results The IP3 content in duodenum homogenate and the expression of Ca M and MLCK in duodenum in model group were significantly higher than those in normal group (P <0.01). Compared with the model group, the IP3 content of Sishenwan high dose group, middle dose group and dextro-treated group decreased (P <0.05 or P <0.01), and the expressions of Ca M and MLCK decreased (P <0.05 or P <0.01). The high dose group of Sishen pill was better than the low dose group (P <0.05), but there was no significant difference between the middle dose of Sishen pill group ). Conclusion The mechanism of the effect of Si Shen Wan on rats with diarrhea due to spleen and kidney yang deficiency may be related to the regulation of IP3-Ca M-MLCK signal pathway in duodenum.