三七总皂苷对糖尿病肾病小鼠的肾保护作用机制研究

Protective effect and mechanism of panax notoginseng saponins on kidney injury in diabetic mice

目的研究三七总皂苷(PNS)对糖尿病肾病小鼠的肾保护作用机制。方法按照体重将雄性C57BL/6小鼠随机分为5组:正常组、模型组和低、中、高3个剂量实验组,每组10只。除正常组以外各组小鼠连续5 d腹腔注射链脲佐菌素50 mg·kg^(-1)建立糖尿病小鼠模型。造模后第2天,实验组灌胃相应剂量药物(PNS:50,100,200 mg·kg^(-1)),正常组和模型组灌胃生理盐水,持续8周。检测血肌酸酐(SCr)和尿素氮(BUN);酶联免疫吸附法检测各组小鼠肾组织谷胱甘肽(GSH)、超氧化物歧化酶(SOD)和血清白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)的含量,蛋白质印迹法测定小鼠肾组织中核因子E-2相关因子(Nrf2)、血红素加氧合酶1(HO-1)蛋白的表达。结果正常组、模型组和低、中、高3个剂量实验组SCr水平分别为(8.24±2.02),(25.10±3.67),(23.41±3.33),(18.81±1.49)和(16.00±2.78)μmol·L^(-1);BUN水平分别为(10.64±2.37),(31.25±1.93),(28.53±3.23),(22.90±2.84)和(18.04±2.94)mmol·L^(-1);SOD水平分别为(32.17±4.48),(13.05±4.90),(18.38±4.91),(23.34±3.46)和(24.65±5.83)U·mg^(-1);GSH水平分别为(13.56±0.71),(4.58±0.66),(5.66±0.34),(7.02±0.22)和(9.21±0.72)mmol·g^(-1);IL-6水平分别为(66.56±13.41),(115.68±7.93),(99.28±20.19),(82.33±9.27)和(76.49±10.14)pg·mL^(-1);TNF-α水平分别为(89.59±15.45),(152.86±26.64),(149.03±17.97),(123.31±13.42)和(111.99±6.49)pg·mL^(-1);Nrf2/β-actin比值分别为0.29±0.14,0.34±0.19,0.70±0.35,0.81±0.34和0.95±0.34;NQO1/β-actin比值分别为0.22±0.12,0.30±0.10,0.45±0.15,0.51±0.16和0.64±0.16。上述指标:模型组与正常组相比,差异均有统计学意义(P<0.05);中、高2个剂量实验组与模型组相比,差异均有统计学意义(P<0.05)。结论PNS对STZ诱导的糖尿病小鼠肾组织损伤具有保护作用,其机制可能与PNS激活Nrf2/HO-1信号通路达到的抗氧化应激、抗炎作用有关。

Objective To study the renal protective effect of panax notoginseng saponins(PNS)in mice with diabetic nephropathy and to explore its mechanism.Methods Male C57BL/6 mice were randomly divided into 5 groups according to weight:normal group,model group,experimental-L group,experimental-M group,experimental-H group,each with 10 mice.The normal mice were injected with normal saline,and the other mice was established diabetic model by injecting 50 mg·kg^(-1)streptozotocin intraperitoneally for 5consecutive days.The three experimental groups were intragastrically administered PNS(50,100,200 mg·kg^(-1)),for 8weeks.The serum creatinine(SCr)and urea nitrogen(BUN)was detected.The content of renal tissue superoxide dismutase(SOD),glutathione(GSH)and serum tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)in mice were determined by enzyme linked immunosorbent assay.The expression of nuclear factor E2-relate factor(Nrf2),Heme oxygenase 1(NQO1)protein in kidney tissue were determined by Western blot.Results The SCr in normal group,model group and low,medium and high dose experimental groups were(8.24±2.02),(25.10±3.67),(23.41±3.32),(18.81±1.49),(16.00±2.78)μmol·L^(-1),respectively;the BUN in the 5 groups were(10.64±2.37),(31.25±1.93),(28.53±3.23),(22.90±2.84),(18.04±2.94)mmol·L^(-1),respectively;the SOD in the 5 groups were(32.17±4.47),(13.05±4.90),(18.38±4.91),(23.34±3.46),(24.65±5.83)U·mg^(-1),respectively;the GSH in the 5 groups were(13.56±0.71),(4.57±0.66),(5.66±0.34),(7.02±0.22),(9.21±0.72)mmol·g^(-1),respectively;the IL-6 in the 5 groups were(66.56±13.41),(115.68±7.93),(99.28±20.19),(82.33±9.27),(76.49±10.14)pg·m L^(-1),respectively;the TNF-αin the 5groups were(89.59±15.45),(152.86±26.64),(149.03±17.97),(123.31±13.42),(111.99±6.49)pg·m L^(-1),respectively;the Nrf2/β-actin ratios in the 5 groups were 0.29±0.14,0.34±0.19,0.70±0.35,0.81±0.34,0.95±0.34,respectively;the NQO1/β-actin ratios in the 5 groups were 0.22±0.12,0.30±0.10,0.45±0.15,0.51±0.16,0.64±0.16,respectively.Comparison between model group and normal group,the difference of the factors were significant(all P<0.05);comparison between medium and high dose experimental groups and model group,the difference of the factors were significant(all P<0.05).Conclusion PNS has a protective effect on STZ-induced renal tissue damage in diabetic mice.The mechanism may be related to the anti-oxidative stress and anti-inflammatory effects of PNS activated by the Nrf2/HO-1 signaling pathway.