CAJ | 학술논문

目的观察臭氧氧化预处理对大鼠肾脏缺血再灌注损伤(IRI)后聚腺苷二磷酸核糖聚合酶-1(PARP-1)/凋亡诱导因子(AIF)通路的作用.方法将30只SD大鼠随机分为假手术组、缺血再灌注组和臭氧氧化预处理组,每组10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:切除右肾后,用无创伤血管夹夹闭左肾动、静脉45 min进行缺血,然后开放血管夹进行再灌注;(3)臭氧氧化预处理组:手术步骤与缺血再灌注组相同,但在术前15d内,每天对该组大鼠经直肠吹入氧气和臭氧的混合气体5.0 ～5.5ml[臭氧浓度为50 mg/L,1.0 mg/(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,免疫组织化学法检测PARP-1/AIF的表达,Western blot检测PARP-1、AIF的含量.结果假手术组、缺血再灌注组和臭氧氧化预处理组的血清肌酐(μmol/L)和尿素氮(mmol/L)分别为19.34±2.27、130.18±23.48、70.91±15.69;7.52±1.98、28.81±10.62、16.54±6.18,苏木素-伊红(HE)染色可见臭氧氧化预处理可以明显减轻因缺血再灌注导致的肾小管上皮细胞变性坏死.与假手术组比较,缺血再灌注组和臭氧氧化预处理组PARP-1、AIF水平均显著升高,而臭氧氧化预处理组的上述指标又明显低于缺血再灌注组.假手术组、缺血再灌注组、臭氧氧化预处理组大鼠肾组织的细胞凋亡指数分别为(3.87±1.50)％、(32.45±4.70)％和(23.47±3.10)％,3组间比较,差异均有统计学意义(P＜0.05).结论臭氧氧化预处理能减轻大鼠肾脏IRI损伤,其机制可能与臭氧氧化预处理抑制细胞凋亡通路PARP-1/AIF的表达有关.
목적 관찰취양양화예처리대대서신장결혈재관주손상(IRI)후취선감이린산핵당취합매-1(PARP-1)/조망유도인자(AIF)통로적작용.방법 장30지SD대서수궤분위가수술조、결혈재관주조화취양양화예처리조,매조10지,진행여하실험:(1)가수술조:절제대서우신후봉합복벽;(2)결혈재관주조:절제우신후,용무창상혈관협협폐좌신동、정맥45 min진행결혈,연후개방혈관협진행재관주;(3)취양양화예처리조:수술보취여결혈재관주조상동,단재술전15d내,매천대해조대서경직장취입양기화취양적혼합기체5.0 ～5.5ml[취양농도위50 mg/L,1.0 mg/(kg·d)].전자동생화분석의검측대서혈청기항화뇨소담,관찰대서신조직적병리개변화세포조망,면역조직화학법검측PARP-1/AIF적표체,Western blot검측PARP-1、AIF적함량.결과 가수술조、결혈재관주조화취양양화예처리조적혈청기항(μmol/L)화뇨소담(mmol/L)분별위19.34±2.27、130.18±23.48、70.91±15.69;7.52±1.98、28.81±10.62、16.54±6.18,소목소-이홍(HE)염색가견취양양화예처리가이명현감경인결혈재관주도치적신소관상피세포변성배사.여가수술조비교,결혈재관주조화취양양화예처리조PARP-1、AIF수평균현저승고,이취양양화예처리조적상술지표우명현저우결혈재관주조.가수술조、결혈재관주조、취양양화예처리조대서신조직적세포조망지수분별위(3.87±1.50)％、(32.45±4.70)％화(23.47±3.10)％,3조간비교,차이균유통계학의의(P＜0.05).결론 취양양화예처리능감경대서신장IRI손상,기궤제가능여취양양화예처리억제세포조망통로PARP-1/AIF적표체유관.
Objective To investigate the effect of ozone oxidative preconditioning (OzoneOP) on poly adenosine diphosphate-ribose polymerase-1 (PARP-1)/apoptosis induce factor (AIF) pathway in rats after acute renal ischemia and reperfusion injury (IRI).Methods Thirty SD rats were randomly divided into three groups:control (sham operation) group,IRI group and OzoneOP group.Each group had 10 rats.In sham operation group,the right kidneys of rats were excised.In IRI group,with no wound vascular clamp,left renal arteries and veins of rats were clamped for 45 min,then vascular clamp opened and the rats were subjected to reperfusion.In OzoneOP group,operation steps were the same as the IRI group,but within 15 d before surgery,the renal OzoneOP was induced by rectal insufflation of oxygen and ozone 5.0-5.5 ml [ozone 50 mg/L,0.5 mg/(kg· d)].The parameters [blood urea nitrogen (BUN) and creatine (Cr)] of renal function were determined by auto-biochemieal analyzer.The expression levels of PARP-1 and AIF were measured by immunohistochemical method,the apoptosis of the nephridial tissue was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL),and the structural changes of renal tissue were examined under a microscope.The expression of PARP-1 and AIF was detected by Western blotting.Results Cr (μmol/L) and BUN (mmol/L) in sham operation,IRI and OzoneOP groups were 19.34 ± 2.27,130.18 ± 23.48,70.91 ± 15.69; 7.52 ± 1.98,28.81 ± 10.62,16.54 ±6.18,respectively.The pathological changes of the renal tissue in IRI group were more serious than those in OzoneOP group.The expression of PARP-1 and AIF was significantly increased in IRI group and OzoneOP group.However,these targets in IRI group were significantly increased as compared with those in OzoneOP group.The apoptosis index in sham operation group,IRI group and OzoneOP group was (3.87 ± 1.50) ％,(32.45 ± 4.70) ％ and (23.47 ± 3.10) ％ respectively (P ＜ 0.05).Conclusion OzoneOP can alleviate renal damage induced by IRI probably by inhibiting the expression of PARP-1/AIF.