吴茱萸次碱对肝肾毒性的初步研究

Preliminary Study on Hepatotoxicity and Nephrotoxicity Induced by Rutaecarpine

目的:探讨吴茱萸次碱Rutecarpine(Rut)在体外对人正常肝细胞(HL7702)、人胚胎肾细胞(HEK293)的影响,采用共培养体系初步比较Rut对肝肾细胞活力的影响,并选用小鼠进行整体验证。方法:①采用MTT法检测Rut在肝肾细胞单独培养或共培养体系中对细胞活力的影响并采用倒置显微镜对细胞形态进行观察;②给予Rut后,检测肝肾细胞培养上清液中丙氨酸转氨酶(ALT),天冬氨酸转氨酶(AST),碱性磷酸酶(ALP),尿素氮(BUN),肌酐(Cr),乳酸脱氢酶(LDH)的变化;③整体实验观察静脉给予Rut对小鼠肝肾功能和组织病理学的影响。结果:①MTT法显示,2,4μg·mL-1的Rut使肝肾细胞活力均下降,且相同浓度下的Rut对肝细胞的抑制作用大于肾细胞;②4μg·mL-1的Rut使肝细胞上清液中的AST,ALP,LDH水平均升高(P<0.01);③整体实验表明,小鼠静脉给予10 mg·kg-1 Rut 7 d后,血清中的ALP水平有极显著的上升(P<0.01),ALT,AST,BUN,Cr水平只出现升高的趋势,病理学检查发现约1/3的小鼠肝脏出现肝细胞核分裂象增多,仅有1只小鼠肾脏出现嗜碱性病变。结论:大剂量Rut可能对肝肾具有一定的毒性作用,肝肾细胞共培养体系与细胞单独培养体系均可用于药物对细胞的毒性研究。

Objective：To study the influence of rutaecarpine（Rut） on human normal hepatic cells（HL7702） and human embryonic kidney cells（HEK293） through in virto experiments.To study the toxicity of rutaecarpine on the vitality of hepatic and renal cells by preliminary compare co-culture system with solo-culture system.Method：①The influence of rutaecarpine on the activity of hepatic and renal cells through MTT method in solo-culture system and co-culture system was detected.② The shape of cells was observed through inverted microscope.③After giving Rut,the change of ALT,AST,ALP,BUN,Cr and LDH in cultural supernatant of hepatic and renal cells was determined.④After giving Rut by intravenous administration,the influence of Rut on hepaticand renal function and histopathology in mice was investigated.Result：①The MTT method showed that 2 μg.mL-1 and 4 μg.mL-1 of Rut decreased the activity of hepatic and renal cells.The inhibitory action on hepatic cells’ activity was higher than that on renal cell activity under the same concentration of Rut.② In cultural supernatant of hepatic cells,4 μg.mL-1 of Rut increased AST,ALP and LDT（P 〈 0.01）.③The animal experiment showed that after giving 10 mg.kg-1 of Rut to mice by intravenous administration for 7 days,ALP in blood serum was increased notably（P 〈 0.01）,while ALT,AST,BUN and Cr had a trend to increase.Histopathological examination revealed that there was an increase in hepatocyte nuclear division in 1/3 mice,meanwhile,basophilic degeneration was only found in one mouse’s kidney.Conclusion：Rut may cause hepatic and renal toxicity.The toxicity to hepatic and kidney cells induced by drugs can be studied by solo-culture system and co-culture system.