不同剂量环磷酰胺对昆明小鼠肝肾组织的损伤作用研究

为研究不同剂量的环磷酰胺对小鼠肝肾组织及功能的损害程度，试验选取40只5周龄健康雌性昆明小鼠，随机分为5组，除对照组外，其余4组小鼠连续3 d分别腹腔注射40、80、120及160 mg/（kg·BW）环磷酰胺溶液，注射后第7天采集肝脏和肾脏组织样品及血清，制备石蜡组织切片及透射电镜切片，观察肝脏和肾脏的组织学变化，检测血清中甘胆酸（CG）、胆碱酯酶（ChE）、血清前白蛋白（PA）、总胆汁酸（TBA）、谷丙转氨酶（ALT）、谷草转氨酶（AST）、尿素氮（BUN）、血清肌酐（SCr）、尿酸（UA）含量/活性。结果显示，环磷酰胺可引起小鼠肝肾组织病理损害，呈剂量依赖性，肝脏比肾脏更早出现损伤。石蜡切片及透射电镜切片观察结果显示，环磷酰胺对肝脏的毒性作用主要表现在肝索紊乱、肝血窦扩张、微胆管淤胆、门管区及中央静脉周围炎性细胞浸润及纤维增生、组织间出血、肝细胞出现脂肪变性及气球样变性，凋亡及坏死细胞增多。肾脏组织损伤主要表现在出血及纤维增生，肾小管上皮细胞胞浆空泡样变、线粒体损伤、核固缩，上皮细胞基底膜增厚。环磷酰胺对膀胱的损害不严重。与对照组相比，40~160 mg/（kg·BW）剂量组小鼠血清中ChE活性及PA水平，120~160 mg/（kg·BW）剂量组AST活性，80~160 mg/（kg·BW）剂量组BUN水平及80 mg/（kg·BW）剂量组UA水平均显著升高（P<0.05）；其余生化指标与对照组差异不显著（P>0.05）。以上结果表明，环磷酰胺注射剂量在80~120 mg/（kg·BW）时可引起昆明小鼠肝脏和肾脏组织及细胞明显的病理损伤。该研究结果可为选择合适的环磷酰胺注射剂量用以制备动物肝脏免疫抑制模型提供试验依据。

Study on the Injury Effect of Different Doses of Cyclophosphamide on the Liver and Kidney Tissues of Kunming Mice

The aim of this study was to study the injury effect of different doses of cyclophosphamide on liver and kidney of mice.40 healthy female Kunming mice aged 5 weeks were randomly divided into 5 groups,injected intraperitoneally with 40,80,120 and 160 mg/(kg·BW)cyclophosphamide for 3 consecutive days,respectively,except the control group.Liver,kidney and serum were collected on 7th day after injection.The paraffin tissue sections and transmission electron microscope sections were prepared,and the histological changes of liver and kidney were observed.The content or activity of CG,ChE,PA,TBA,ALT,AST,BUN,SCr,UA in serum were detected.The results showed that cyclophosphamide could cause pathological injury to liver and kidney of mice in a dose-dependent manner,and the liver showed injury earlier than the kidney.The observed results of paraffin sections and transmission electron microscopy sections showed that the toxic effects of cyclophosphamide on the liver were mainly manifested in hepatic cord disorders,hepatic sinusoidal dilatation,microbiliary cholestasis,inflammatory cell infiltration and fibrous proliferation in the portal area and around the central vein,hemorrhage between tissues,fatty degeneration and balloon-like degeneration of liver cells,apoptosis and necrotic cells increased.Kidney tissue damages were mainly manifested in hemorrhage and fibrosis,renal tubular epithelial cell cytoplasmic vacuolation,mitochondrial damage,nuclear pyknosis,and thickening of the basement membrane of epithelial cells.Cyclophosphamide did not cause serious damage to the bladder.Compared with the control group,serum ChE activites and PA contents in 40-160 mg/(kg·BW)groups,AST activities in 120-160 mg/(kg·BW)groups,BUN contents in 80-160 mg/(kg·BW)groups,and UA level in 80 mg/(kg·BW)group were increased significantly(P<0.05).The other biochemical indicators were not significantly different from the control group(P>0.05).The results showed that cyclophosphamide injection at a dose of 80-120 mg/(kg·BW)could cause obvious pathological damage to liver and kidney tissues and cells in Kunming mice.The results of the study could provide an experimental basis for the selection of appropriate cyclophosphamide in establishment of animal liver immunosuppression model.