石榴皮鞣质对肾小球硬化大鼠内源性物质代谢的影响及代谢通路分析

Effects of tannins from Granati Pericarpium on endogenous substance metabolism and analysis on metabolic pathway in rats with glomeruloscerosis

目的利用高效液相色谱串联质谱(HPLC-MS)的尿液代谢组学研究方法,考察石榴皮鞣质对肾小球硬化大鼠内源性物质代谢的影响,寻找潜在的生物标志物并分析其代谢途径,为研究石榴皮鞣质对肾小球硬化大鼠的改善作用及其机制提供理论依据。方法将48只SD大鼠随机分为假手术组,模型组,贝那普利组,石榴皮鞣质高、中、低剂量组,采用单侧肾切除结合2次尾iv盐酸多柔比星的方法制备肾小球硬化大鼠模型,ig给药8周,每周收集大鼠24 h尿液,用HPLC-MS对其检测分析;采用主成分分析(PCA)、偏最小二乘法判别分析(PLS-DA)及正交偏最小二乘法判别分析(OPLS-DA)等方法对各组大鼠尿液代谢物进行聚类分析,筛选出潜在的生物标志物并构建相应的代谢通路。结果代谢组学分析发现给药后第8周各组大鼠尿液有明显的聚类现象,通过对重要变量的分析鉴定,筛选出10个潜在的生物标志物;涉及的代谢通路包括色氨酸、丙氨酸、天冬氨酸和谷氨酸等多种氨基酸的代谢通路。结论石榴皮鞣质干预下的肾小球硬化大鼠内源性代谢物聚类性趋近正常水平,为进一步阐明石榴皮鞣质对肾小球硬化大鼠的作用机制提供依据。

Objective A urinary metabonomics method based on liquid chromatography-mass spectrometry （HPLC-MS） was used to inspect the effect of tannins from Granati Pericarpium on endogenous metabolism of rats with glomeruloscerosis, the potential biomarkers were searched and the metabolic pathways were analyzed to provide relevant theory for treatment of glomeruloscerosis. Methods Forty-eight SD rats were divided into control group, model group, benazepril hydrochloridec group, and three doses of pomerganate tannis groups. Glomerular sclerosis model was established by unilateral nephrectomy combined with two tail iv adriamycin; 24 h urine of rats was collected after the rats were orally given tannins from Granati Pericarpium for 8 weeks; HPLC-MS was used for testing and analyzing the urine; Principal component analysis （PCA）, partial least squares-discriminant analysis （PLS-DA） and orthogonal partial least squares discriminant analysis （OPLS-DA） were used for data treatment of each group to search the potential biomarkers and build the metabolicomics pathways. Results The urinary metabolism of rats was clustering. Ten potential biomakers were screened by analysis and identification of important variables; The metabonomics pathways involved tryptophan, alanine, aspartate, glutamate metabolism, etc. Conclusion The trend of endogenous metabolism of rats with glomeruloscemsis is normal after intervened by tannis in pomegranate tannins from Granati Pericarpium. It provides the basement with the further research of clarififing the mechanism of tannins from Granati Pericarpium inpacting on rats with glomeruloscerosis.