广防己的肾毒性及代谢组学研究

Nephrotoxicity study of Aristolochia fangchi in rats by metabonomics

目的:采用代谢组学方法研究大鼠予广防己灌胃后尿液代谢产物的改变情况。方法:雄性SD大鼠64只随机分为广防己组和正常对照组,每组32只。广防己组大鼠予广防己8.1g/(kg.d)灌胃,正常对照组灌服等体积蒸馏水,连续灌胃4周。在灌胃前,灌胃2周、4周及停药2周后收集大鼠24h尿液,利用核磁共振(nuclear magnetic resonance,NMR)技术测定大鼠尿液代谢产物1HNMR谱,进行模式识别分析;取血检测血尿素氮(blood urea nitrogen,BUN)和血清肌酐(serum creatinine,SCr)含量;取肾脏组织行病理学检查。结果:与正常对照组比较,给药2周时广防己组大鼠血BUN含量明显升高(P<0.05);肾组织出现肾小管细胞肿胀,肾小球球囊结构破坏,肾间质炎性细胞浸润。给药4周时,BUN和SCr含量显著升高(P<0.05),肾组织病理改变加重。停药恢复2周后,广防己组BUN含量仍较正常对照组明显升高(P<0.05),且病理改变与给药4周时无明显差异。大鼠尿样代谢谱经主成分分析提示,广防己组与正常对照组在各时间点代谢物图谱均有差异,整个给药期间,广防己组柠檬酸含量持续下降,牛磺酸含量持续上升。在给药2周时,伴马尿酸盐含量上升;给药4周时伴马尿酸盐、2-酮戊二酸含量下降,氧化三甲胺和肌酐/肌酸含量上升;停药恢复2周后肌酐/肌酸和2-酮戊二酸含量下降,马尿酸盐和氧化三甲胺含量上升。结论:10倍药典剂量广防己给药2周可造成肾功能损害,随给药时间延长,损伤加重,停药后有一定恢复。广防己可能还具有一定的肝脏毒性作用。

Objective： To study the changes of metabolites in rat urine after treatment of Aristolochia fangchi decoction by metabonomic method. Methods： Sixty-four male SD rats were divided into Aristolochia fangchi group and normal control group. Rats in the Aristolochia fangchi group were orally administered with 8.1 g/（kg · d） of Aristolochia fangchi and the normal control group was administered with equal volume of distilled water for 4 weeks. Twenty-four hour urine was collected at different time points （before, after 2- and 4-week administration and 2 weeks after administration） and their ^1H nuclear magnetic resonance （NMR） spectra were acquired and subjected to data process, including principal component analysis （PCA）, partial least squares discriminant analysis （PLS-DA） or orthogonal signal correction （QSC）. The contents of blood urea nitrogen （BUN） and serum creatinine （SCr） and histopathological changes of the renal tissues were also detected. Results： The content of BUN of the Aristolochia fangchi group was markedly higher than that of the normal control group after 2-week administration （P〈0. 05）, and cellular edema in tubular endothelial cells, structure damage of glomeruli and inflammatory cell infiltration were found in the kidney. Along with the lasting of administration to 4-week, the renal injury in the Aristolochia fangchi group became more serious, and the contents of BUN and SCr were all significantly higher as compared with the normal control group （P〈0.05）. Two weeks after administration, the content of BUN in the Aristolochia fangchi group was still higher than that in the normal control group （P〈0.05）, and the pathological changes in renal tissues were not different from those on the 4th week. The urine of the Aristolochia fangchi group could be readily discriminated from the normal control group at every time point based on PCA. During the whole administration period, compared with the normal control group, the concentration of urinary taurine was increased time-dependently, while the citrate was decreased in the Aristolochia fangchi group. The concentration of hippurate was increased at the 2nd week and the 6th week （2 weeks after drug withdrawal） but decreased at the 4th week; the concentration of creatinine/creatine was increased at the 4th week but decreased at the 6th week; the concentration of 2-oxo-glutarate was decreased and the concentration of trimethylamine oxide was increased at the 4th and 6th week. Conclusion： High-dose Aristolochia fangchi can induce renal lesion and its seriousness is correspondent to the lasting of administration. Aristolochia fangchi may also have toxicity on liver.