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目的研究果糖饮食及皮下注射内毒素(LPS)致大鼠肝胰岛素抵抗(IR)及内毒素血症中,LPS对肝细胞线粒体功能的影响。方法 30只SD雄性大鼠随机分为三组。对照组(NC组):普通饲料喂养;果糖组(HFD组):10%果糖水喂养;LPS组:皮下注射LPS 300μg/(kg·d)。8周糖耐量实验后,检测血浆肝酶、空腹胰岛素、LPS变化,计算IR指数;检测肝组织氧化损伤及能量代谢指标;Western blot检测肝组织胰岛素信号转导蛋白及线粒体内膜蛋白(UCP2)表达;观察肝组织病理学变化。分离培养大鼠肝细胞分为四组。NC组:DMEM培养液培养;HFD组:培养液+果糖水(4.5 g/L);LPS组:培养液+LPS(10 mg/L);果糖+LPS组(H+L组):培养液+果糖水(4.5 g/L)+LPS(10 mg/L)。20 h后,检测肝细胞线粒体功能及胰岛素信号转导蛋白表达。结果与NC组比较,HFD组与LPS组LPS、肝酶及氧化损伤产物显著升高,能量代谢异常(P<0.01),胰岛素信号转导蛋白表达降低,UCP2表达升高(P<0.01);HFD组与LPS组上述指标变化差异无统计学意义(P>0.05)。HFD组、LPS组和H+L组细胞上清液氧化损伤产物高于NC组(P<0.05或P<0.01),能量代谢异常(P<0.05),胰岛素信号转导蛋白表达下降,UCP2表达升高(P<0.05或P<0.01);各干预组之间以上指标变化差异无统计学意义(P>0.05)。结论果糖饮食及皮下注射LPS致大鼠肝IR中,伴有内毒素血症。LPS可促发肝氧化应激,影响肝细胞线粒体功能,促进代谢性疾病的发生。
Objective To investigate the effect of LPS on the mitochondrial function of hepatocytes induced by fructose diet and subcutaneous injection of endotoxin (LPS) in rats with hepatic insulin resistance (IR) and endotoxemia. Methods Thirty SD male rats were randomly divided into three groups. The control group (NC group): ordinary feed; fructose group (HFD group): 10% fructose water feeding; LPS group: subcutaneous injection of LPS 300μg / (kg · d). The levels of hepatic enzymes, fasting insulin and LPS were measured after 8-week glucose tolerance test, and the IR index was calculated. The oxidative damage and energy metabolism index of liver tissue were detected; Western blot was used to detect the expression of insulin signal transduction protein and mitochondrial inner membrane protein (UCP2) Expression; observed liver histopathological changes. Rat hepatocytes were isolated and divided into four groups. NC group: DMEM culture medium; HFD group: culture medium + fructose water (4.5 g / L); LPS group: culture medium + LPS (10 mg / L); fructose + LPS group + Fructose (4.5 g / L) + LPS (10 mg / L). Twenty hours later, mitochondrial function and insulin signal transduction protein expression in hepatocytes were detected. Results Compared with NC group, LPS, liver enzymes and products of oxidative damage in HFD group and LPS group were significantly increased, with abnormal energy metabolism (P <0.01), decreased insulin signal transduction protein expression and increased UCP2 expression (P <0.01). There was no significant difference in the above indexes between HFD group and LPS group (P> 0.05). The oxidative damage products in HFD, LPS and H + L groups were higher than those in NC group (P <0.05 or P <0.01), the energy metabolism was abnormal (P <0.05), the expression of insulin signal transduction protein (P <0.05 or P <0.01). There was no significant difference in the above indexes among the intervention groups (P> 0.05). Conclusion Fructose diet and subcutaneous injection of LPS induced liver IR in rats with endotoxemia. LPS can promote liver oxidative stress, affect the mitochondrial function of liver cells and promote the occurrence of metabolic diseases.