马钱子总生物碱致肾毒性模型建立及甘草内生菌代谢产物减毒作用对比

Establishment of Nephrotoxicity Model Induced by Total Alkaloids of STRYCHNI SEMEN and Comparison on Attenuation Effects of Endophytes Metabolites from GLYCYRRHIZAE RADIX ET RHIZOMA

目的:建立马钱子总生物碱(TAS)致肾毒性模型,探究甘草及其内生菌代谢产物对TAS致肾毒性模型的减毒作用。方法:Wistar大鼠随机分为正常对照组、模型对照组、甘草提取物176 mg/kg组、甘草内生真菌17、33 mg/kg组、甘草内生细菌17、33 mg/kg组,每组10只,以7.50 mg/mL的TAS建立大鼠肾毒性模型,灌胃给予甘草和甘草内生菌进行干预,观察大鼠体征状况,检测血清尿素氮(BUN)、肌酐(Cr)含量,计算脏器指数,观察组织病理学变化及损伤分数等指标,评价TAS肾毒性模型及甘草内生菌对TAS致肾毒性的减毒作用。结果:与正常对照组比较,模型对照组血清BUN、Cr含量及脏器指数明显升高(P<0.05或P<0.01),肾脏组织中肾小管空泡样变形,肾小管萎缩,且肾小球肾小管损伤评分明显升高(P<0.01);与模型对照组比较,甘草提取物及其内生菌代谢物各组大鼠体征状况逐渐恢复,BUN的含量显著降低(P<0.01),大鼠肾组织病理损伤减轻,肾小球与肾小管损伤评分降低(P<0.05或P<0.01);与甘草提取物176 mg/kg组比较,甘草内生细菌代谢物17 mg/kg组BUN含量显著升高(P<0.01),甘草内生真菌代谢物33 mg/kg组、甘草内生细菌代谢物33 mg/kg组肾小管损伤评分和肾小球损伤评分明显降低(P<0.05或P<0.01)。结论:7.50 mg/kg的TAS成功建立大鼠的肾毒性模型;甘草内生菌代谢物具有减毒作用,一定剂量内,甘草内生真菌代谢物呈现出优于甘草的减毒作用。

Objective:To establish a nephrotoxicity model induced by total alkaloids of STRYCHNI SEMEN(TAS) and explore the attenuation effects of GLYCYRRHIZAE RADIX ET RHIZOMA and its endophytes Metabolites on the nephrotoxicity model induced by TAS. Methods:Male Wistar rats were randomly assigned into a blank control group, a model control group, a GLYCYRRHIZAE RADIX ET RHIZOMA extract(176 mg/kg) group, GLYCYRRHIZAE RADIX ET RHIZOMA endophytic fungi(17 mg/kg and 33 mg/kg) groups, and GLYCYRRHIZAE RADIX ET RHIZOMA endophytic bacteria(17 mg/kg and 33 mg/kg) groups, with ten rats in each group. TAS(7.50 mg/mL) was used to establish the rat model of nephrotoxicity. After intervention with GLYCYRRHIZAE RADIX ET RHIZOMA extract and endophytes, the physical signs of rats were observed and the serum levels of blood urea nitrogen(BUN) and creatinine(Cr) were determined. Organ index was calculated and the histopathological changes were observed and scored, which were adopted to evaluate the attenuation effects of GLYCYRRHIZAE RADIX ET RHIZOMA extract and endophytes on the rat nephrotoxicity induced by TAS. Results:Compared with the blank control group, the modeling elevated the serum BUN and Cr levels and increased the organ index(P<0.05 or P<0.01). Moroever, it led to the vacuolation and shrinking of renal tubules and increased the glomerular and tubular damage scores(P<0.01). Compared with the model control group, GLYCYRRHIZAE RADIX ET RHIZOMA extract and endophytes made the physical signs of rats recover, decreased the BUN level(P<0.01), alleviated the pathological damage of renal tissue, and decreased the glomerular and tubular damage scores(P<0.05 or P<0.01). Compared with the GLYCYRRHIZAE RADIX ET RHIZOMA extract(176 mg/kg), GLYCYRRHIZAE RADIX ET RHIZOMA endophytic bacteria(17 mg/kg) increased the the BUN level(P<0.01) and GLYCYRRHIZAE RADIX ET RHIZOMA endophytic fungi(33 mg/kg) and bacteria(33 mg/kg) decreased the glomerular and tubular damage scores(P<0.05 or P<0.01). Conclusion:The rat model of nephrotoxicity was successfully established with 7.50 mg/kg TAS. The endophye from GLYCYRRHIZAE RADIX ET RHIZOMA showed attenuation effect on the nephrotoxicity. Moreover, the endophytic fungi outperformed GLYCYRRHIZAE RADIX ET RHIZOMA extract in the attenuation effect within a certain dose range.