甘草酸拮抗雷公藤多苷所致肾毒性的分子机制研究

  目的  从转运体角度探究甘草酸拮抗雷公藤多苷所致肾毒性的分子机制。  方法  采用网络药理学预测雷公藤多苷和甘草酸作用于肾毒性的靶点, 从中选出与肾脏转运体有关的靶点, 并通过动物实验对其进行验证。采用ELISA法测定大鼠血清中BUN和Scr含量, 通过HE染色判断大鼠肾脏病理情况, 采用qPCR和Western blot检测转运体基因和蛋白表达量。  结果  网络药理学预测雷公藤多苷和甘草酸共同作用于肾毒性的靶点73个, 分子功能81种, 主要富集于蛋白结合, 涉及转运体ABCB1和ABCC2。动物实验表明, 雷公藤多苷组大鼠肾脏的BUN和Scr含量显著升高, 病理结果显示大鼠肾脏受到较严重的损伤; 配伍甘草酸后BUN和Scr含量显著降低, 肾脏损伤得以改善。qPCR和Western blot结果显示雷公藤多苷可显著下调P-gp和MRP2基因和蛋白表达量; 配伍甘草酸后P-gp和MRP2基因和蛋白的表达量显著上调。  结论  甘草酸通过影响P-gp和MRP2转运体, 加速毒性物质排出, 拮抗雷公藤多苷所致肾毒性, 为临床两药联合治疗肾病、减轻毒性反应提供实验依据。 

Research on the Molecular Mechanism for Glycyrrhizic Acid against Tripterygium Glycosides-Induced Nephrotoxicity

  OBJECTIVE  To explore the molecular mechanism of glycyrrhizic acid antagonizing nephrotoxicity induced by tripterygium glycosides from the perspective of transporters.  METHODS  The targets of tripterygium glycosides and glycyrrhizic acid on nephrotoxicity were predicted by network pharmacology, and the targets related to renal transporters were selected and verified by animal experiments. The contents of BUN and Scr in rat serum were measured by ELISA, the pathological condition of rat kidney was detected by HE staining, and the expressions of transporter gene and protein were detected by qPCR and Western blot.  RESULTS  Network pharmacology predicted that there were 73 targets and 81 molecular functions of the two drugs on nephrotoxicity, which were mainly concentrated in protein binding, involving transporters ABCB1 and ABCC2. Animal experiments showed that tripterygium glycosides significantly increased the contents of BUN and Scr in rat kidney, the renal pathological results showed that the kidney was severely injured; Compatibility of tripterygium glycosides and glycyrrhizic acid significantly reduced the contents of BUN and Scr and alleviated kidney damage. qPCR and Western blot showed that tripterygium glycosides significantly down-regulated the gene and protein expressions of P-gp and MRP2; Compatibility of tripterygium glycosides and glycyrrhizic acid significantly up-regulated the gene and protein expressions of P-gp and MRP2.  CONCLUSION  Glycyrrhizic acid can accelerate the excretion of toxic substances and antagonize the nephrotoxicity caused by tripterygium glycosides by affecting P-gp and MRP2 transporters. This study provides experimental evidence for the combination of two drugs in clinical treatment of nephropathy and alleviation of toxic reactions.