纳米银诱导长爪沙鼠肝细胞凋亡与血清生化指标的研究

Research on nano silver induced gerbil hepatocyte apoptosis

目的探索经口灌胃纳米银建立肝细胞凋亡动物模型,并初步阐明其发生机制。方法选用2～3月龄长爪沙鼠,随机分成3个剂量试验组和1个生理盐水阴性对照组,每组均为14只,雌性各半,分笼饲养。3个剂量组分别每天按30、125和500 mg/kg体重给予各组小鼠各剂量受试物,对照组小鼠经口灌胃同批号生理盐水,剂量为25 ml/kg,连续28 d。注射结束后第3天,经小鼠眼眶采血,检测小鼠血清超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)水平,用TUNEL法、流式细胞技术检测肝细胞凋亡情况,并进行光子显微镜、电子显微镜观察,探索导致明显急性肝损伤的可能机制。结果与对照组相比,剂量125和500 mg/kg的受试组血清中SOD活力和GSH-Px活力显著降低(P<0.01),MDA含量显著提高(P<0.01);30mg/kg剂量受试组血清SOD和GSH-Px活力降低,MDA含量提高,但差异不显著(P>0.05);组织病理学检测可见典型的凋亡小体结构;流式细胞术、TUNEL法检测及电镜检测都见明显的凋亡小体。结论注射125 mg/kg剂量以上纳米银的老鼠肝细胞出现了明显的凋亡。

Objective To explore the establishment of intraperitoneal injection of silver nanoparticles on hepatic cell apoptosis in animal models,and clarify its pathogenesis.Methods Fifty-six 2-3 month Mongolian gerbils were randomly divided into 4 groups with 14 replications per group.The treatment 4 groups were handled by peritoneal injection with 0,30,125 and 500 mg respectively for 28 days.Mice from the blank control group received identical procedure,with the exception of drug applicaion.The 3rd days after the injection,blood was collected for biochemical analysis.The apoptosis was assessed by measuring the content of MDA and the activity of SOD and GSH.The mechanism led to marked acute liver damage was explored by light microscopy and electron microscopy.Results Compared with normal control group,the activity of SOD and GSH significantily decreased（P＆lt;0.01） and MDA level significanly increased（P＆lt;0.01）in the serum of the dose of 125 mg/kg and 500 mg groups.There was such a trend that the SOD and GSH activity decreased with the increase level of MDA in the serum of 30 mg/kg group,but the discrepancy was not significant（P＆gt;0.05）.Histopathologic examination revealed a visible typical apoptotic body structure.Flow cytometry,TUNEL assay and electron microscopic detection olso showed apparent apoptotic bodies.Conclusion The injection of 125 mg/kg dose above nano silver mouse liver cells appeared obvious apoptosis.