七叶皂苷钠对小鼠肾缺血再灌注损伤的保护作用

Protective effect of aescin sodium on renal ischemia-reperfusion injury in mice

目的 研究七叶皂苷钠对小鼠肾缺血再灌注损伤(IRI)的保护作用。方法 将雄性8周龄的WTC57BL/6小鼠,分为假手术组、模型组和低、高剂量实验组。假手术组充分麻醉后,沿小鼠腹正中线打开腹腔暴露双肾后充分游离双侧肾蒂,并按0.5 mL/10 g剂量注射0.9%NaCl溶液,缝合腹壁(即仅游离而不夹闭双侧肾);模型组实验过程与假手术组相同,但游离双侧肾后,用无损伤微型动脉夹彻底夹闭双肾肾蒂35 min后恢复血流灌注(夹闭双侧肾蒂但不给药);低、高剂量实验组过程与缺血再灌注损伤模型组相同,但需要在缺血35 min后1 h内腹腔注射1、2 mg·kg^(-1)七叶皂苷钠,后续在术后6、12、18 h按时按量分次腹腔注射七叶皂苷钠溶液。术后24 h用全自动生化分析仪检测血清肌酸酐(Scr)、尿素氮(BUN)水平;用试剂盒检测肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)、超氧化物歧化酶(SOD)、丙二醛(MDA)含量;在光镜下观察肾组织病理学改变。结果 假手术组、模型组和低、高剂量实验组血清Scr分别为(16.36±1.32)、(133.24±8.69)、(111.46±7.18)、(98.32±8.41)μmol·L^(-1);BUN分别为(7.12±1.02)、(90.77±3.62)、(74.51±2.95)、(52.64±3.30)mmol·L^(-1);血清TNF-α分别为(1.01±0.04)、(7.37±1.01)、(2.40±0.75)、(2.24±0.73)pg·mL^(-1);血清IL-10分别为(1.08±0.12)、(7.61±1.73)、(2.25±0.85)、(2.11±0.53)pg·mL^(-1);血清SOD分别为(160.40±7.69)、(95.43±4.10)、(141.11±5.87)、(138.82±2.28)U·mg^(-1)prot;血清MDA分别为(12.61±1.35)、(17.23±1.55)、(15.11±1.04)、(13.73±0.80)nmol·mg^(-1)prot。与假手术组相比,模型组小鼠血清Scr、BUN、TNF-α、IL-10、MDA水平以及肾病理损伤评分均升高(均P<0.05),SOD水平降低(P<0.05);与模型组相比,低、高剂量实验组小鼠血清Scr、BUN、TNF-ɑ、IL-10、MDA水平均降低(均P<0.05),SOD水平升高(P<0.05)。结论 七叶皂苷钠能减轻小鼠肾缺血再灌注损伤,其机制可能与减轻氧化应激和炎症反应有关。

Objective To investigate the protective effect of aescin sodium on renal ischemia-reperfusion injury (IRI) in mice.MethodsEight-week-old male WTC57BL/6 mice were divided into 4 groups:sham operation group,model group,experimental-L group and experimental-H group.After full anesthesia in sham operation group,the abdominal cavity was opened along the median line of the mouse abdomen to expose both kidneys and fully free both renal pedicles,and0.9%Na Cl solution was injected at the dose of 0.5 m L/10 g to suture the abdominal wall (only free but not clamp both kidneys).The experimental procedure of the model group was the same as that of the sham operation group,but after freeing both kidneys,blood perfusion was restored after the pedicle of both kidneys was completely clamped with non-damaging mini-artery clamp for 35 min (the pedicle of both kidneys was clamped but no drug was administered).The procedure of the experimental-L group and experimental-H group was the same as that of the ischemia reperfusion injury model group,but 1 and 2 mg·kg^(-1)sodium aesculin was injected intraperitoneally within 1 hour after 35 min of ischemia,and then intraperitoneally injected sodium aesculin solution in time and quantity at 6,12 and 18 h after surgery.At 24h after operation,the automatic biochemical analyzer detected serum creatinine (Scr) and blood urea nitrogen (BUN)levels;the kit was use to detect the contents of tumor necrosis factor-α (TNF-α),interleukin-10 (IL-10),superoxide dismutase (SOD) and malondialdehyde (MDA);the pathological changes of renal tissue were observed under light microscope.Results The serum Scr in sham operation group,model group,experimental-L group and experimental-H group were (16.36±1.32),(133.24±8.69),(111.46±7.18),(98.32±8.41)μmol·L^(-1);the serum BUN were (7.12±1.02),(90.77±3.62),(74.51±2.95),(52.64±3.30) mmol·L^(-1);the serum TNF-α were (1.01±0.04),(7.37±1.01),(2.40±0.75),(2.24±0.73) pg·m L^(-1);serum IL-10 were(1.08±0.12),(7.61±1.73),(2.25±0.85),(2.11±0.53) pg·m L^(-1);the serum SOD were(160.40±7.69),(95.43±4.10),(141.11±5.87),(138.82±2.28) U·mg^(-1)prot;the serum MDA were(12.61±1.35),(17.23±1.55),(15.11±1.04),(13.73±0.80) nmol·mg^(-1)prot.Compared with sham operation group,the levels of serum Scr,BUN,TNF-α,IL-10,MDA in model group were increased (all P0.05),and the level of SOD was decreased (P0.05);compared with model group,the levels of serum Scr,BUN,TNF-ɑ,IL-10,MDA and renal pathological scores in experimental-L,-H groups were significantly decreased (all P0.05),and the level of SOD was increased (P0.05).Conclusion Sodium aescinate can alleviate renal ischemia-reperfusion injury in mice,and its mechanism may be related to reducing oxidative stress and inflammatory response.