CAJ | 학술논문

目的研究大鼠局灶性脑缺血再灌注损伤后多聚腺苷二磷酸核糖聚合酶(PARP)和凋亡诱导因子(AIF)在海马CAI区的表达,探讨西洛他唑预处理能否通过PARP/AIF途径发挥脑保护作用. 方法将135只雄性SD大鼠按随机数字表法分为3组:假手术组、模型组、西洛他唑组,每组45只.采用线栓法阻塞大鼠大脑中动脉制作局灶性脑缺血再灌注损伤模型.西洛他唑组造模前灌胃给予30 mg/kg剂量西洛他唑(2次).每组根据再灌注时间点不同(6 h、24 h、72 h)分为3个亚组,每亚组15只.应用TUNEL法检测神经细胞凋亡变化,Western blotting法检测AIF、腺苷二磷酸核糖(PAR)在不同时间点的变化,RT-PCR法检测AIF mRNA的表达变化. 结果大鼠局灶性脑缺血再灌注损伤后出现AIF核移位.与假手术组比较,模型组凋亡细胞数明显增加,AIF、PAR含量及AIF mRNA表达明显增加,24 h时最显著,差异均有统计学意义(P＜0.05).西洛他唑组再灌注6h、24 h、72 h各亚组凋亡细胞数较模型组中各亚组明显减少,AIF、PAR含量较模型组各亚组明显降低,AIF mRNA表达亦明显减少,24h时最显著,差异有统计学意义(P＜0.05). 结论西洛他唑对大鼠脑缺血再灌注损伤有一定保护作用,其抗神经细胞凋亡的机制之一可能是通过抑制脑缺血损伤引起的PARP的过度活化及AIF的易位而实现的.
목적 연구대서국조성뇌결혈재관주손상후다취선감이린산핵당취합매(PARP)화조망유도인자(AIF)재해마CAI구적표체,탐토서락타서예처리능부통과PARP/AIF도경발휘뇌보호작용. 방법 장135지웅성SD대서안수궤수자표법분위3조:가수술조、모형조、서락타서조,매조45지.채용선전법조새대서대뇌중동맥제작국조성뇌결혈재관주손상모형.서락타서조조모전관위급여30 mg/kg제량서락타서(2차).매조근거재관주시간점불동(6 h、24 h、72 h)분위3개아조,매아조15지.응용TUNEL법검측신경세포조망변화,Western blotting법검측AIF、선감이린산핵당(PAR)재불동시간점적변화,RT-PCR법검측AIF mRNA적표체변화. 결과 대서국조성뇌결혈재관주손상후출현AIF핵이위.여가수술조비교,모형조조망세포수명현증가,AIF、PAR함량급AIF mRNA표체명현증가,24 h시최현저,차이균유통계학의의(P＜0.05).서락타서조재관주6h、24 h、72 h각아조조망세포수교모형조중각아조명현감소,AIF、PAR함량교모형조각아조명현강저,AIF mRNA표체역명현감소,24h시최현저,차이유통계학의의(P＜0.05). 결론 서락타서대대서뇌결혈재관주손상유일정보호작용,기항신경세포조망적궤제지일가능시통과억제뇌결혈손상인기적PARP적과도활화급AIF적역위이실현적.
Objective To study the expressions of poly(ADP-ribose) polymerase (PARP) and apoptosis-inducing factor (AIF) in the hippocampal CA1 region of rats after focal cerebral ischemia reperfusion injury,and elucidate the neuroprotective effect ofcilostazol pretreatment through PARP/AIF pathway.Methods One hundred and thirty-five male SD rats were randomly divided into three groups (n=45) for experiments:sham-operated group,ischemia/reperfusion (I/R) vehicle group and cilostazol pretreatment group.Embolization thread was inserted into all rats except those in the sham-operated group for 2 hours to establish middle cerebral artery occlusion models; and then,reperfusion for 6,24,72 hours was performed,and rats of each group were sub-divided into three groups according to reperfusion times (n=l 5).Cilostazol pretreatment group was given cilostazol (30 mg/kg,twice orally) before surgery.The apoptosis cells of the hippocampus were detected by Terminal deoxynucleotidyl Transferasemediated dUTP nick end-labeling (TUNEL).Western blotting was used to test the dynamic changes of AIF and polyadenosine diphosphate ribose (PAR) levels at different times.The mRNA expression of AIF was assessed by real time-PCR.Results There appeared subsequent translocation of AIF after focal cerebral ischemia reperfusion injury in rats.As compared with those in the sham-operated group,the apoptotic cells statistically increased; AIF and PAR contents and AIF mRNA expression significantly increased in the I/R vehicle group reaching its peak level at 24 h ofreperfusion (P＜0.05).As compared with of the I/R model subgroups,the cilostazol pretreatment subgroups had significantly decreased AIF and PAR contents,AIF mRNA expression and number of apoptosis cells (P＜0.05); 24 h of reperfusion subgroup had the most obvious effect (P＜0.05).Conclusion Cilostazol pretreatment in rats after ischemia-reperfusion injury has certain neroprotective effect,whose resistance roles in nerve cell apoptosis may be implemented by inhibiting excessive PARP activation and AIF translocation caused by cerebral ischemia injury.