苯丙胺对大鼠肝脏毒性损害的实验研究

发布时间：2018-08-31 08:18

【摘要】： 目的：本研究通过苯丙胺(amphetamine，AP)诱导的大鼠中毒模型，以其对机体肝脏的毒性损害为切入点，观察在不同时间和不同剂量下，大鼠肝组织和细胞的形态学变化以及脂肪酸合成酶的活性表达，探讨AP对机体脏器损害的毒理作用。方法： SD大鼠65只，体重200±20g，随机分成对照组(Z组，n=5)和实验组(A、B、C，n=20)，饲养及光照节律正常。A组为AP小剂量组，0.5mg／kg；B组为中剂量组，，2.5mg／kg；C组为大剂量组，5mg／kg；Z组为生理盐水组，0.5mg／kg；给药方式为左后肢肌肉注射。每天上午给药一次。各组分别在用药14d、28d和42d处死，取肝并经石蜡包埋、切片。 1．应用HE染色学方法，观察大鼠肝小叶结构及肝细胞脂肪变性、空泡样变性、肝细胞核固缩现象； 2．应用免疫组织化学方法定性和定位大鼠肝脂肪酸合成酶阳性细胞变化； 3．应用Western Blot方法定量检测大鼠肝脂肪酸合成酶量的变化。结果： 1．HE染色后，光镜下观察到，大鼠肝内肝窦变窄、消失，肝索解离、结构消失；肝细胞肿胀、浑浊、细胞浆出现脂滴，较大的脂滴将细胞核挤压变形；肝细胞呈现局部、部分或弥漫性脂肪变性；严重时可见肝细胞气球样变，边界不清，可见核固缩。毒性损害严重程度随着时间的延长和剂量的增加而加重，呈正相关性。 2．免疫组化结果显示：大鼠肝内脂肪酸合成酶阳性细胞胞浆呈黄染，阳性细胞数随时间和剂量的增加而减少，呈负相关。 3．Western Blot结果表明大鼠肝内脂肪酸合成酶的含量随着时间和剂量的增加而减少。结论： AP对肝脏有明显的毒性作用，主要表现为肝细胞的脂肪变性和细胞水肿；同时，经免疫组化和蛋白印迹方法证明，脂肪酸合成酶的细胞数目和含量随着AP的使用时间和剂量的增加而减少。
[Abstract]:Objective: to study the toxic effects of amphetamine (amphetamine,AP) on the liver of rats at different time and dosage. The morphologic changes of liver tissues and cells and the expression of fatty acid synthase activity in rat liver were studied to investigate the toxicological effects of AP on organ damage. Methods: 65 SD rats, weighing 200 卤20 g, were randomly divided into two groups: control group (group Z, n = 5) and experimental group (group A, n = 20). The feeding and illumination rhythm was normal. Group A was fed with AP (0.5 mg 路kg ~ (-1) 路kg ~ (-1) and the middle dose group was (2.5 mg 路kg ~ (-1) 路L ~ (-1) 路L ~ (-1) 路L ~ (-1) 路kg ~ (-1). Group C was treated with 5 mg 路kg ~ (-1) of normal saline group, and 0.5 mg 路kg ~ (-1) 路kg ~ (-1) of normal saline group, and the method of administration was intramuscular injection to the left hind limb. The medicine is given once a day in the morning. Each group was killed at 14d and 42d respectively. Liver was taken and embedded in paraffin wax. 1. The structure of hepatic lobule and hepatic cell fatty degeneration and vacuolar degeneration were observed by HE staining. The changes of hepatic fatty acid synthase positive cells were determined by immunohistochemical method. 3. The content of fatty acid synthase in rat liver was quantitatively detected by Western Blot method. Results: after 1.HE staining, it was observed that the hepatic sinusoids became narrow, disappeared, the hepatic cord dissociated and the structure disappeared, the liver cells were swollen, turbid, and lipid droplets appeared in the cytoplasm of the liver. Large lipid droplets deform the nucleus; hepatocytes present local, partial or diffuse steatosis; in severe cases, hepatocyte balloon degeneration is seen, the boundary is unclear, and nuclear pyknosis can be seen. The severity of toxicity increased with time and dose. 2. Immunohistochemical results showed that the cytoplasm of fatty acid synthase positive cells in rat liver was yellow staining. The results of 3.Western Blot showed that the content of fatty acid synthase decreased with the increase of time and dose. Conclusion: AP has obvious toxic effects on liver, mainly manifested as steatosis and edema of hepatocytes, and was proved by immunohistochemistry and Western blotting. The cell number and content of fatty acid synthase decreased with the increase of time and dose of AP.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R363

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