CAJ | 학술논문

目的探讨凋亡相关基因p53在自发性高血压大鼠(SHR)视网膜缺血再灌注损伤(RIR)后视网膜毛细血管细胞凋亡中的作用.方法将60只SHR随机分为假手术组(SHR-SH)和缺血组(SHR-RIR),每组各30只大鼠,每组又按照不同再灌注时间分别再分为再灌注后2、6、24、72 h和7 d共5个小组,每小组各6只大鼠.选取60只Wistar-Kyoto大鼠(WKY)同样分为假手术组(WKY-SH)和缺血组(wKY-IIR)作为对照.建立大鼠RIR损伤动物模型,采用末端脱氧核苷酸转移酶介导的dUTP缺口翻译法(TUNEL)法观察大鼠视网膜毛细血管细胞凋亡,通过免疫组织化学链酶卵白素-生物素复合体法(SP)检测RIR后不同时段视网膜组织中p53基因的表达变化.结果 SHR视网膜毛细血管细胞凋亡率在RIR后2、6、24,72 h和7 d时分别为(8.64±0.56)%、(14.92±0.99)%、(24.72±2.98)%、(16.53±1.80)%和(7.12±1.10)%.SHR视网膜毛细血管在RIR后2 h p53表达即开始增加,6 h继续增多,24 h达到高峰,72 h仍维持在较高水平,7 d时仍有少量表达,与SHR-SH比较,差异有统计学意义(P<0.01).WKY-SH和SHR-SH组各时间点细胞凋亡和p53表达无明显差异.WKY-RIR与SHR-RIR相应时间点比较,差异有统计学意义(P<0.01).结论 p53可能通过诱导或促进细胞凋亡参与大鼠RIR;高血压状态下发生RIR视网膜毛细血管细胞凋亡更为严重,且尤以缺血再灌注后24 h为著.
목적 탐토조망상관기인p53재자발성고혈압대서(SHR)시망막결혈재관주손상(RIR)후시망막모세혈관세포조망중적작용.방법장60지SHR수궤분위가수술조(SHR-SH)화결혈조(SHR-RIR),매조각30지대서,매조우안조불동재관주시간분별재분위재관주후2、6、24、72 h화7 d공5개소조,매소조각6지대서.선취60지Wistar-Kyoto대서(WKY)동양분위가수술조(WKY-SH)화결혈조(wKY-IIR)작위대조.건립대서RIR손상동물모형,채용말단탈양핵감산전이매개도적dUTP결구번역법(TUNEL)법관찰대서시망막모세혈관세포조망,통과면역조직화학련매란백소-생물소복합체법(SP)검측RIR후불동시단시망막조직중p53기인적표체변화.결과 SHR시망막모세혈관세포조망솔재RIR후2、6、24,72 h화7 d시분별위(8.64±0.56)%、(14.92±0.99)%、(24.72±2.98)%、(16.53±1.80)%화(7.12±1.10)%.SHR시망막모세혈관재RIR후2 h p53표체즉개시증가,6 h계속증다,24 h체도고봉,72 h잉유지재교고수평,7 d시잉유소량표체,여SHR-SH비교,차이유통계학의의(P<0.01).WKY-SH화SHR-SH조각시간점세포조망화p53표체무명현차이.WKY-RIR여SHR-RIR상응시간점비교,차이유통계학의의(P<0.01).결론 p53가능통과유도혹촉진세포조망삼여대서RIR;고혈압상태하발생RIR시망막모세혈관세포조망경위엄중,차우이결혈재관주후24 h위저.
Objective To investigate the effect of apoptosis-related gene p53 on the apoptosis of retinal capillary cells in rats with spontaneously hypertensive (SHR) after ischemic reperfusion injury. Methods A total of 60 SHR rats were randomly divided into sham group (SHR-SH) and retinal ischemie reperfusion group (SHR-RIR), which were subdivided into 5 subgroups according to the time after RIR: 2, 6, 24, and 72 hours and 7 days, with 6 rats in each subgroup. Another 60 Wistar-Kyoto (WKY) rats were divided into the same groups as the SHR rats as the control. The RIR model was set up. The apoptosis of retinal capillary cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) methods and the expression of p53 was determined by streptavidin-perosidase (SP) immunohistochemistry. Results The apoptosis rate of retinal capillary cells in the 5 SHR-RIR groups was (8.64±0.56)%, (14.92±0.99)%, (24.72±2.98)%, (16.53±1.80)%, and (7.12±1.10)%,respectively. The expression of p53 in SHR-RIR groups increased at the 2nd hour after RIR, reached the peak at the 24th hour, kept the high level at the 72nd hour, and remained a little at the 7th day, which was significantly different from which in the SHR-SH groups (P<0. 01). The expression of p53 were higher in SHR-IR groups than that in the WKY-RIR groups (P<0. 01). Conclusions p53 may play a part in RIR injury by inducing or promoting apoptosis. The apoptosis of retinal capillary cells after RIR is more severe under the hypertension, and reaches the peak at the 24 hour after RIR.