【摘要】 目的 探讨梅花鹿鹿茸总蛋白提取物（SVPr）对糖尿病肾病（DN）大鼠肾损伤的治疗作用及分子机制。方法 将120只Sprague Dawley大鼠按照简单随机数字表法分为对照组、模型组、二甲双胍组、低剂量SVPr组、中剂量SVPr组、高剂量SVPr组，每组20只。模型组、二甲双胍组、低剂量SVPr组、中剂量SVPr组、高剂量SVPr组大鼠造模前禁食12 h,单次腹腔注射链脲佐菌素50 mg·kg^-1制备DN大鼠模型；造模成功后，二甲双胍组大鼠给予二甲双胍500 mg·kg^-1·d^-1灌胃，低剂量SVPr组、中剂量SVPr组、高剂量SVPr组大鼠分别经尾静脉注射质量浓度为1.47、2.94、4.41 g·L^-1的SVPr溶液0.1 mL,对照组及模型组大鼠注射等量生理盐水，每日1次，持续给药4周。采用生物化学法检测各组大鼠血尿素氮（BUN）、血肌酐（SCr）、血糖、24 h尿蛋白水平，酶联免疫吸附试验法检测各组大鼠肾组织中活性氧（ROS）、核因子-κB （NF-κB）水平，Western blot法检测各组大鼠肾组织中沉默调节蛋白1（SIRT1）、核因子E2相关因子2 （Nrf-2）、血红素加氧酶-1（HO-1）蛋白相对表达量。结果 模型组大鼠BUN、SCr、24 h尿蛋白、血糖水平均显著高于对照组（P<0.05）;低剂量SVPr组、中剂量SVPr组、高剂量SVPr组和二甲双胍组大鼠BUN、SCr、24 h尿蛋白、血糖水平显著低于模型组（P<0.05）;高剂量SVPr组大鼠SCr、24 h尿蛋白水平显著低于二甲双胍组（P<0.05）,高剂量SVPr组与二甲双胍组大鼠BUN、血糖水平比较差异无统计学意义（P>0.05）;中剂量SVPr组大鼠SCr水平显著低于二甲双胍组，BUN、血糖水平显著高于二甲双胍组（P<0.05）;中剂量SVPr组与二甲双胍组大鼠24 h尿蛋白水平比较差异无统计学意义（P>0.05）;低剂量SVPr组大鼠BUN、24 h尿蛋白、血糖水平显著高于二甲双胍组（P<0.05）,低剂量SVPr组与二甲双胍组大鼠SCr水平比较差异无统计学意义（P>0.05）。高剂量SVPr组大鼠BUN、血糖、24 h尿蛋白、SCr水平显著低于中剂量SVPr和低剂量SVPr组，中剂量SVPr组BUN、血糖、24 h尿蛋白、SCr水平显著低于低剂量SVPr组（P<0.05）。模型组大鼠肾组织中ROS、NF-κB水平显著高于对照组（P<0.05）;低剂量SVPr组、中剂量SVPr组、高剂量SVPr组和二甲双胍组大鼠肾组织中ROS、NF-κB水平显著低于模型组（P<0.05）;高剂量SVPr组大鼠肾组织中ROS、NF-κB水平显著低于二甲双胍组（P<0.05）;中剂量SVPr组大鼠肾组织中ROS水平显著高于二甲双胍组（P<0.05）,中剂量SVPr组与二甲双胍组大鼠肾组织中NF-κB水平差异无统计学意义（P>0.05）;低剂量SVPr组大鼠肾组织中ROS、NF-κB水平显著高于二甲双胍组（P<0.05）;高剂量SVPr组大鼠肾组织中ROS、NF-κB水平显著低于中剂量SVPr组和低剂量SVPr组；中剂量SVPr组大鼠肾组织中ROS水平显著低于低剂量SVPr组（P<0.05）,中剂量SVPr组与低剂量SVPr组大鼠肾组织中NF-κB水平比较差异无统计学意义（P>0.05）。模型组大鼠肾组织中SIRT1、Nrf-2、HO-1相对表达量均显著低于对照组（P<0.05）;低剂量SVPr组、中剂量SVPr组、高剂量SVPr组和二甲双胍组大鼠肾组织中SIRT1、Nrf-2、HO-1相对表达量显著高于模型组（P<0.05）;高剂量SVPr组大鼠肾组织中SIRT1、Nrf-2相对表达量显著高于二甲双胍组（P<0.05）,高剂量SVPr组与二甲双胍组大鼠肾组织中HO-1相对表达量比较差异无统计学意义（P>0.05）;中剂量SVPr组大鼠肾组织中SIRT1、Nrf-2相对表达量显著高于二甲双胍组，HO-1相对表达量显著低于二甲双胍组（P<0.05）;低剂量SVPr组大鼠肾组织中SIRT1相对表达量显著高于二甲双胍组（P<0.05）,低剂量SVPr组与二甲双胍组大鼠肾组织中Nrf-2、HO-1相对表达量比较差异无统计学意义（P>0.05）;高剂量SVPr组大鼠肾组织中SIRT1、Nrf-2、HO-1相对表达量显著高于中剂量SVPr组和低剂量SVPr组，中剂量SVPr组大鼠肾组织中SIRT1、Nrf-2、HO-1相对表达量显著高于低剂量SVPr组（P<0.05）。结论 SVPr可通过上调SIRT1表达来抑制NF-κB转录活性、激活HO-1/Nrf-2抗氧化应激信号，减轻DN大鼠肾损伤。更多还原

【Abstract】 Objective To investigate the therapeutic effect and molecular mechanism of Sika deer velvet antler protein（SVPr） on renal injury in diabetes nephropathy（DN） rats.Methods A total of 120 Sprague Dawley rats were divided into control group, model group, metformin group, low-dose of SVPr group, medium-dose of SVPr group and high-dose of SVPr group according to the simple random number table, with 20 rats in each group.The rats in the model group, metformin group, low-dose of SVPr group, medium-dose of SVPr group and high-dose of SVPr group were fasted for 12 hours before modeling, and the rats were single intraperitoneally injected with streptozocin 50 mg·kg^-1 to prepare the DN model.After successful modeling, the rats in the metformin group were given metformin by gavage with a dose of 500 mg·kg^-1·d^-1;the rats in the low-dose of SVPr group, medium-dose of SVPr group and high-dose of SVPr group were injected via tail vein with 0.1 mL SVPr solution with concentrations of 1.47,2.94 and 4.41 g·L^-1,respectively; the rats in the control group and model group were injected with the same amount of normal saline, once a day for 4 weeks.The levels of blood urea nitrogen（BUN）,serum creatinine（SCr）,blood glucose and 24 h urine protein of rats in each group were detected by biochemical method, and the levels of reactive oxygen species（ROS） and nuclear factor-κB（NF-κB） in kidney tissue of rats in each group were detected by enzyme-linked immunosorbent assay, the relative expressions of silencing regulatory protein 1（SIRT1）,nuclear factor erythroid-2 related factor 2（Nrf-2）,heme oxygenase-1（HO-1） protein in renal tissues of rats in each group were detected by Western blot.Results The levels of BUN,SCr, 24 h urine protein and blood glucose of rats in the model group were significantly higher than those in the control group（P<0.05）;the levels of BUN,SCr, 24 h urine protein and blood glucose of rats in the low-dose of SVPr group, medium-dose of SVPr group, high-dose of SVPr group and metformin group were significantly lower than those in the model group（P<0.05）;the levels of SCr and 24 h urine protein of rats in the high-dose of SVPr group were significantly lower than those in the metformin group（P<0.05）,and there was no significant difference in the levels of BUN and blood glucose of rats between the high-dose of SVPr group and metformin group（P>0.05）;the level of SCr of rats in the medium-dose of SVPr group was significantly lower than that in the metformin group, the levels of BUN and blood glucose of rats were significantly higher than those in the metformin group（P<0.05）;there was no significant difference in the level of 24 h urine protein of rats between the medium-dose of SVPr group and metformin group（P>0.05）;the levels of BUN,SCr and 24 h urine protein of rats in the low-dose of SVPr group were significantly higher than those in the metformin group（P<0.05）,and there was no significant difference in the level of SCr of rats between the low-dose of SVPr group and metformin group（P>0.05）.The levels of BUN,blood glucose, 24 h urine protein and SCr of rats in the high-dose of SVPr group were significantly lower than those in the medium-dose of SVPr group and low-dose of SVPr group（P<0.05）;and the levels of BUN,blood glucose, 24 h urine protein and SCr of rats in the medium-dose of SVPr group were significantly lower than those in the low-dose of SVPr group（P<0.05）.The levels of ROS and NF-κB in kidney tissue of rats in the model group were significantly higher than those in the control group（P<0.05）;the levels of ROS and NF-κB in renal tissue of rats in the low-dose of SVPr group, medium-dose of SVPr group, high-dose of SVPr group and metformin group were significantly lower than those in the model group（P<0.05）;the levels of ROS and NF-κB in renal tissue of rats in the high-dose of SVPr group were significantly lower than those in the metformin group（P<0.05）;the level of ROS in kidney tissue of rats in the medium-dose of SVPr group was significantly higher than that in the metformin group（P<0.05）,there was no significant difference in NF-κB level in kidney tissue of rats between the medium-dose of SVPr group and metformin group（P>0.05）;the levels of ROS and NF-κB in kidney tissue of rats in the low-dose of SVPr group were significantly higher than those in the metformin group（P<0.05）;the levels of ROS and NF-κB in renal tissue of rats in the high-dose of SVPr group were significantly lower than those in the medium-dose of SVPr group and low-dose of SVPr group（P<0.05）;the level of ROS in kidney tissue of rats in the medium-dose of SVPr group was significantly lower than that in the low-dose of SVPr group（P<0.05）,there was no significant difference in NF-κB level in kidney tissue of rats between the medium-dose of SVPr group and low-dose of SVPr group（P>0.05）.The relative expression levels of SIRT1,Nrf-2 and HO-1 in kidney tissue of rats in the model group were significantly lower than those in the control group（P<0.05）;the relative expression levels of SIRT1,Nrf-2 and HO-1 in kidney tissue of rats in the low-dose of SVPr group, medium-dose of SVPr group, high-dose of SVPr group and metformin group were significantly higher than those in the model group（P<0.05）;the relative expression levels of SIRT1 and Nrf-2 in kidney tissue of rats in the high-dose of SVPr group were significantly higher than those in the metformin group（P<0.05）.There was no significant difference in the relative expression level of HO-1 in kidney tissue of rats between the high-dose of SVPr group and metformin group（P>0.05）;the relative expression levels of SIRT1 and Nrf-2 in kidney tissue of rats in the medium-dose of SVPr group were significantly higher than those in the metformin group, and the relative expression level of HO-1 was significantly lower than that in the metformin group（P<0.05）;the relative expression level of SIRT1 in kidney tissue of rats in the low-dose of SVPr group was significantly higher than that in the metformin group（P<0.05）,and there was no significant difference in the relative expression levels of Nrf-2 and HO-1 in kidney tissue of rats between the low-dose of SVPr group and metformin group（P>0.05）;the relative expression levels of SIRT1,Nrf-2 and HO-1 in kidney tissue of rats in the high-dose of SVPr group were significantly higher than those in the medium-dose of SVPr group and low-dose of SVPr group（P<0.05）;the relative expression levels of SIRT1,Nrf-2 and HO-1 in kidney tissue of rats in the medium-dose of SVPr group were significantly higher than those in the low-dose of SVPr group（P<0.05）.Conclusion SVPr can reduce kidney damage of DN rats through inhibiting NF-κB transcriptional activity and activating HO-1/Nrf-2 antioxidant stress signal by up-regulating SIRT1 expression.更多还原