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目的:采用高脂饲料喂养16周诱导NAFLD大鼠模型,对其肝细胞脂质蓄积及氧化应激状态进行评价。方法:30只雄性SD大鼠随机分正常组和模型组,每组15只。正常组予基础饲料喂养,模型组予高脂饲料喂养。16周后,比较两组大鼠的肝湿重、Lee’s指数;超声下观察大鼠肝脏影像学改变;检测血清甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白(LDL-C)、高密度脂蛋白(HDL-C)、肝组织匀浆TC、TG含量;取肝组织行HE染色、油红O染色和透射电镜观察组织学改变;Ⅳ型胶原酶法提取原代肝细胞行油红O染色并检测肝细胞一氧化氮(NO)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)和超氧化物歧化酶(SOD)含量。结果:模型组肝脏超声所见符合脂肪性肝病影像学改变;与正常组比较,模型组大鼠肝湿重、Lee’s指数、血清TC、TG、LDL-C,肝组织匀浆TC、TG及肝细胞MDA、NO明显升高(P<0.01),而血清HDL-C及肝细胞SOD、GSH-Px水平明显降低(P<0.01);肝组织及肝细胞病理提示肝细胞脂质蓄积。结论:高脂饲料喂养16周的大鼠模型其肝细胞存在脂质蓄积和氧化应激状态,是成功的NAFLD大鼠模型。
OBJECTIVE: To induce NAFLD rat model by feeding with high-fat diet for 16 weeks and evaluate its lipid accumulation and oxidative stress in liver cells. Methods: Thirty male SD rats were randomly divided into normal group and model group, with 15 rats in each group. Normal group fed with basal diet, model group fed with high fat diet. After 16 weeks, the liver wet weight and Lee’s index of the two groups were compared; the liver changes were observed under ultrasound; the contents of serum triglyceride (TG), total cholesterol (TC), low density lipoprotein (LDL-C ), High density lipoprotein (HDL-C), liver tissue homogenate TC, TG content; take the liver tissue HE staining, oil red O staining and transmission electron microscopy histological changes; type Ⅳ collagenase extraction of primary hepatocytes Oil red O staining and detection of liver cell nitric oxide (NO), malondialdehyde (MDA), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) content. Results: Compared with the normal group, the liver wet weight, Lee’s index, serum TC, TG, LDL-C, liver homogenate TC, TG and liver (P <0.01), while the level of serum HDL-C and the level of SOD and GSH-Px in hepatocytes were significantly decreased (P <0.01). The liver tissue and liver cell pathology showed the accumulation of lipid in hepatocytes. Conclusion: The rat model of lipid peroxidation induced by high-fat diet for 16 weeks is a successful rat model of NAFLD.