二神丸中药物炮制前后对肠道菌群和UCP2基因表达的影响

目的:观察二神丸中补骨脂、肉豆蔻炮制前后对脾肾阳虚泄泻模型大鼠肠道菌群和肾脏线粒体解偶联蛋白2(UCP2)基因表达的影响,探讨二神丸中药物炮制增效的机制。方法:将24只SD雄性大鼠随机等分为正常组、模型组、二神丸生品组(补骨脂生品+肉豆蔻生品)和二神丸炮制品组(盐补骨脂+煨肉豆蔻),运用复合造模方法复制大鼠脾肾阳虚泄泻模型,后二组灌胃给药量3.5 g·kg~(-1),正常组和模型组给予同等体积生理盐水。采用实时荧光定量聚合酶链式反应技术检测各组肠道菌群,研究二神丸对肠道菌群的影响;使用免疫组化染色SP法测定各组大鼠肾脏组织UCP2基因表达。结果:脾肾阳虚泄泻大鼠肠道菌群和UCP2含量与造模前相比发生明显变化,其中长双歧杆菌、乳酸杆菌菌群异常低下,大肠埃希菌、粪肠球菌含量显著增加,肾脏组织UCP2含量明显升高。给予二神丸后,对肠道菌群失调和UCP2异常均有明显的调节和改善作用,尤以炮制组(盐补骨脂+煨肉豆蔻)效果为最佳。结论:由盐炙补骨脂和麸煨肉豆蔻组方的二神丸促进脾肾阳虚泄泻模型有益菌的增殖、抑制有害菌以及下调肾脏组织UCP2效果更加明显,提示其增效可能与调节肠道菌群及机体基础代谢率相关。

Effect of Chinese Medicine in Ershen Wan Before and After Being Processed on Intestinal Flora and mRNA Expression of Uncoupling Protein 2

Objective: To observe effect of Psoraleae Fructus and Myristicae Semen in Ershen Wan after and before being processed on intestinal flora and expression levels of uncoupling protein 2 (UCP2) mRNA of spleen-kidney Yang deficiency diarrhea rats, to preliminarily explore its possible mechanism. Method: Rat model of spleen-kidney Yang deficiency diarrhea was made through perfusion of adenine, irregular diet and reception of ice Sennae Folium extract.Rats were averagely subdivided into four groups, such as normal group, model group, raw products group and processing group of Ershen Wan.Real-time fluorescence quantitative technique was employed to investigate effect of Ershen Wan on intestinal flora.Expression levels of UCP2 mRNA in renal tissues were assayed by immunohistochemical staining SP method. Result: Intestinal flora and expression levels of UCP2 mRNA of spleen-kidney Yang deficiency diarrhea rats changed obviously by comparing with before modeling;flora of Bifidobacterium longum and Lactobacillus was abnormally low, contents of Escherichia coli and Enterococcus faecalis increased significantly, the expression level of UCP2 mRNA in rat renal tissues of model group was significantly increased.The alteration of intestinal flora and expression levels of UCP2 mRNA recovered after receiving Ershen Wan, and the processing group was the best. Conclusion: Ershen Wan, consisting of salt-fried Psoraleae Fructus and bran-roasted Myristicae Semen, has function of promoting proliferation of intestinal available bacteria, inhibiting intestinal harmful bacteria and down-regulating expression levels of UCP2 mRNA in renal tissues, which may be related to regulation of intestinal flora and the body''s basal metabolic rate.