益智仁提取物对糖尿病肾病小鼠肾氧化应激损伤的影响及机制研究

Effect and Mechanism of Alpinia Oxyphylla Extract on Oxidative Stress Injury of Kidney in Diabetic Nephropathy Mice

【目的】探讨益智仁提取物对糖尿病肾病(DN)小鼠肾氧化应激损伤的影响及机制。【方法】将40只SPF级SD雄性小鼠随机分为对照组、模型组及益智仁提取物低、高剂量组,每组10只。低剂量组、高剂量组及模型组成功建立DN模型后,同时间分别给予5 g/(kg?d)、20 g/(kg?d)益智仁提取物溶液、等量生理盐水灌胃处理(连续4周),对照组于同时间给予等量生理盐水灌胃处理。采用全自动生化分析仪测定24 h尿蛋白、血肌酐、血尿素氮水平;采用苏木精-伊红(HE)染色观察肾脏组织病理变化;采用比色法测定超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量,采用ELISA法检测谷胱甘肽(GSH-px)含量;采用免疫印迹法测定肾组织中核因子E2相关因子2/血红素加氧酶-1(Nrf2/HO-1)通路蛋白Nrf2、HO-1水平。【结果】DN模型建立成功;与对照组比较,模型组及低剂量组、高剂量组大鼠24 h尿蛋白、血尿素氮、血肌酐水平均升高,SOD、GSH-px水平及Nrf2、HO-1蛋白水平均降低(P<0.05);与模型组比较,低剂量组、高剂量组大鼠24 h尿蛋白、尿素氮、血肌酐水平均降低,SOD、GSH-px水平及Nrf2、HO-1蛋白水平均升高(P<0.05);与低剂量组比较,高剂量组大鼠24 h尿蛋白、血尿素氮、血肌酐水平均升高,SOD、GSH-px水平及Nrf2、HO-1蛋白水平均降低(P<0.05)。【结论】益智仁提取物对DN小鼠肾脏发挥保护作用,可能是通过促进Nrf2/HO-1通路活化抑制肾脏氧化应激损伤实现。

【Objective】To explore the effect of alpinia oxyphylla extract on oxidative stress injury of kidney in diabetic nephropathy mice,and to analyze the possible mechanism.【Methods】Forty SPF male SD mice were randomly divided into the control group,the model group,low and high dose group of alpinia oxyphylla extract(with 10 mice in each group).After establishing diabetic nephropathy mouse model(DN mouse),5 g/(kg?d)and 20 g/(kg?d)of alpinia oxyphylla extract solution with certain amount of normal saline were given to the low and high dose group mice for 4 weeks;while DN mice in the control group was given the same amount of normal saline at the same time period.Hematoxylin-eosin(HE)staining was used to observe the renal histopathology.The levels of 24 h urine protein,serum creatinine and urea nitrogen were determined by automatic biochemical analyzer,the activity of Superoxide dismutase,the content of malondialdehyde(MDA)and the content of glutathione peroxidase(GSH-px)were determined by colorimetry and ELISA,respectively.Nrf2 and HO-1 levels of nuclear factor E2 related factor 2 Heme oxygenase-1(Nrf2/HO-1)were measured by Western blotting.【Results】Compared to SD mice in the control group,the levels of 24 h urinary protein,urea nitrogen and serum creatinine in the DN model group and mice with the low and high dose groups of alpinia oxyphylla extract increased,while the levels of SOD,GSH-px and Nrf2,HO-1 protein decreased(P<0.05).Compared with the DN model group,the levels of 24 h urinary protein,urea nitrogen and serum creatinine decreased in the low-and high-dose groups,and the levels of SOD,GSH-px and Nrf2,HO-1 protein increased(P<0.05).Levels of urine protein,urea nitrogen and serum creatinine in mice with the low-dose group of alpinia oxyphylla extract were lower than those in the high-dose group,while the levels of SOD,GSH-px and Nrf2,HO-1 protein were lower(P<0.05).【Conclusion】The protective effect of Alpinia Oxyphylla extract on kidney of DN mice may be realized by promoting activation of Nrf2/HO-1 pathway and inhibiting oxidative stress injury of kidney.