石榴皮鞣质对肾纤维化大鼠内源性代谢物的影响

Effects of tannins in pomegranate bark on endogenous metabolites in rats with renal fibrosis

目的:运用代谢组学方法研究石榴皮鞣质对肾纤维化大鼠内源性代谢物的影响,寻找潜在的生物标志物并分析代谢途径,为肾纤维化的治疗和石榴皮鞣质的利用提供相关理论依据。方法:SD大鼠随机分为5组,假手术组,模型组,缬沙坦组,石榴皮鞣质高剂量组、低剂量组。除假手术组,其余各组采用左侧输尿管结扎方法构建肾纤维化模型,假手术组的输尿管不结扎和剪断。收集各组大鼠术后3周尿样,经预处理后,进行液相色谱-高分辨质谱分析,获得总离子流图。对LC-MS所得数据进行预处理,导入SIMCA-P软件进行主成分分析(PCA)和正交信号校正偏最小二乘法分析(OPLS),根据获得的得分图、载荷图的分析结果,寻找潜在的生物标记物。结果:代谢组学分析发现造模后第3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13个潜在的生物标记物。结论:石榴皮鞣质干预下的肾纤维化大鼠内源性代谢物聚类性趋近正常水平,代谢组学分析法寻找潜在生物标志物可行且有效。

OBJECTIVE To investigate the effects of tannins from pomegranate bark on endogenous metabolites in rats with renal fibrosis by using metabonomics method, find potential biomarkers and analyze the metabolizing pathway, and provide rele- vant theoretical background on the treatment of renal fibrosis and utilization of tannins from pomegranate bark. METHODS SD rats were randomly divided into five groups, i. e. , sham operation group, model group, valsartan group, high dose and low dose groups of tannins from pomegranate bark. Except for sham operation group,left ureter of rat was ligated to construct renal fi- brosis model, while in sham operated group, ureter would not be ligated or cut off. In the five groups, urine were collected from rats in Week 3 and analyzed by liquid chromatography-high resolution mass spectrometry （LC-MS） to obtain total ion current. Data from LC-MS were pretreated, and imported into SIMCA-P to perform principal component analysis （PCA） and orthogonal signal correction partial least squares analysis （OPLS）,and potential biomarkers would be found according to the results of the score and load charts. RESULTS The result of metabolomics analysis showed that the urine of rats was clustering since the third week after model was constructed. Through analysis and identification of important variances, 13 potential biomarkers ware screened out. CONCLUSION Distribution of endogenous metaholites of rats with renal fibrosis tends to be the normal lev- el after intervention of tannins from pomegranate bark. It is feasible and effective to find potential biomarkers by metabolomics method.