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目的:探讨臭氧后处理对大鼠肾脏缺血再灌注损伤(IRI)中缺氧诱导因子-1α(HIF-1α)及肾小管上皮细胞凋亡的影响。方法:随机将36只SD大鼠分为假手术组、缺血再灌注组和臭氧后处理组,每组12只,进行如下实验:①假手术组:切除大鼠右肾后缝合腹壁;②缺血再灌注组:切除大鼠右肾后,用无创伤血管夹夹闭左肾动、静脉45min进行缺血,然后开放血管夹进行再灌注;③臭氧后处理组:手术步骤与缺血再灌注组相同,但在术后10d内,每天对该组大鼠经直肠吹入氧气和臭氧的混合气体2.5-3.0ml[臭氧浓度为50mg/L,0.5mg/(kg·d)]。全自动生化分析仪检测大鼠血清肌酐和尿素氮,观察大鼠肾组织的病理改变和细胞凋亡情况,蛋白免疫印迹法(Western blot)检测各组大鼠HIF-1α的含量。结果:与假手术组相比,缺血再灌注组和臭氧后处理组血清尿素氮、肌酐水平均显著升高,而缺血再灌注组的尿素氮、肌酐表达量又明显高于臭氧后处理组。臭氧后处理组的肾组织病理学改变较缺血再灌注组轻。假手术组、缺血再灌注组、缺血后处理组大鼠肾组织的细胞凋亡指数分别为4.32±1.84、39.28±4.06和27.42±2.83,3组间比较,差异均有统计学意义(P<0.05)。Western Blot结果显示,在假手术组、缺氧再灌注组和臭氧后处理组中HIF-1α表达量依次增加,差异有统计学意义(P<0.05)。结论:臭氧后处理可能通过显著增加HIF-1α的表达而抑制肾小管上皮细胞的凋亡,从而发挥肾脏保护作用。
Objective: To investigate the effect of ozone postconditioning on the apoptosis of hypoxia inducible factor-1α (HIF-1α) and renal tubular epithelial cells in renal ischemia-reperfusion injury (IRI) rats. Methods: 36 SD rats were randomly divided into sham operation group, ischemia reperfusion group and ozone postconditioning group, with 12 rats in each group. The following experiments were performed: ① Sham-operation group: the right kidney was resected and the abdominal wall was resected; ② Ischemia / reperfusion group: After excision of the right kidney in rats, the left renal artery and vein were closed with an atraumatic vessel for 45 min for ischemia, and then the vessel clamp was opened for reperfusion; ③ Ozone postconditioning group: Perfused group was the same, but 2.5-3.0ml [ozone concentration 50mg / L, 0.5mg / (kg · d)] was injected into the rectum once a day for 10 days. Serum creatinine and urea nitrogen were detected by automatic biochemical analyzer. The pathological changes and apoptosis of renal tissue were observed. The content of HIF-1α in each group was detected by Western blot. Results: Compared with the sham group, the levels of serum urea nitrogen and creatinine were significantly increased in the ischemia-reperfusion group and the ozone-treated group, while the levels of urea nitrogen and creatinine in the ischemia-reperfusion group were significantly higher than those in the ozone group group. The histopathological changes in the ozone-treated group were lighter than those in the ischemia-reperfusion group. The apoptosis index of kidney in sham-operated group, ischemia-reperfusion group and ischemic postconditioning group were 4.32 ± 1.84, 39.28 ± 4.06 and 27.42 ± 2.83, respectively, the difference was statistically significant ( P <0.05). The results of Western Blot showed that the expression of HIF-1α in sham operation group, hypoxia reperfusion group and ozone postconditioning group increased in turn, with statistical significance (P <0.05). Conclusion: Ozone postconditioning may play a protective role in renal tubular epithelial cells by increasing the expression of HIF-1α significantly.