CAJ | 학술논문

目的探讨激肽释放酶(kallikrein)对脑梗死周边区域缺血神经细胞凋亡的作用.方法建立大鼠大脑中动脉闭塞(MCAO)模型,将造模成功的30只大鼠采用随机数字表法分为以下几组:A组,空白对照组;B组,注射生理盐水;C组,注射pAdCMV-人类组织激肽释放酶(HTK):每组10只.观察每组大鼠处理前后神经功能损害评分(NSS),用免疫组化技术检测外源性HTK的表达,并通过TUNEL染色、RT-PCR检测细胞凋亡及凋亡因子bcl-2、bax、caspase-3 mRNA水平.结果治疗后24h在C组中可见HTK表达阳性细胞并逐渐增多,72h后达高峰.与B组以及A组相比差异有统计学意义(112±6.1、68±4.2、59±3.9,P<0.05).治疗后72h,C组大鼠NSS神经功能评分明显低于B组以及A组(6.70±0.16、8.13±0.16、7.93±0.20,P<0.05);治疗后7 d差异更明显(5.14±0.18、7.82±0.14、7.91±0.10,P<0.01).凋亡神经细胞集中于梗死周边区,治疗后72h及7d时,C组平均TUNEL阳性细胞数较B组与A组明显减少(72 h:10.1±0.9、16.7±1.1、20.4±0.8;7 d:15.2±1.2、33.6±1.3、28.8±1.7,P<0.05).与B、A组比较,C组中bc1.2 mRNA水平增高不明显(P>0.05);治疗后72 h及7 d,bax与caspase.3 mRNA水平则明显降低(P<0.05). 结论 Kallikrein可能通过减少凋亡因子bax、caspase-3的表达,抑制脑梗死周边区域的神经细胞凋亡,从而达到保护缺血神经细胞,改善神经功能的作用.
목적 탐토격태석방매(kallikrein)대뇌경사주변구역결혈신경세포조망적작용.방법 건립대서대뇌중동맥폐새(MCAO)모형,장조모성공적30지대서채용수궤수자표법분위이하궤조:A조,공백대조조;B조,주사생리염수;C조,주사pAdCMV-인류조직격태석방매(HTK):매조10지.관찰매조대서처리전후신경공능손해평분(NSS),용면역조화기술검측외원성HTK적표체,병통과TUNEL염색、RT-PCR검측세포조망급조망인자bcl-2、bax、caspase-3 mRNA수평.결과 치료후24h재C조중가견HTK표체양성세포병축점증다,72h후체고봉.여B조이급A조상비차이유통계학의의(112±6.1、68±4.2、59±3.9,P<0.05).치료후72h,C조대서NSS신경공능평분명현저우B조이급A조(6.70±0.16、8.13±0.16、7.93±0.20,P<0.05);치료후7 d차이경명현(5.14±0.18、7.82±0.14、7.91±0.10,P<0.01).조망신경세포집중우경사주변구,치료후72h급7d시,C조평균TUNEL양성세포수교B조여A조명현감소(72 h:10.1±0.9、16.7±1.1、20.4±0.8;7 d:15.2±1.2、33.6±1.3、28.8±1.7,P<0.05).여B、A조비교,C조중bc1.2 mRNA수평증고불명현(P>0.05);치료후72 h급7 d,bax여caspase.3 mRNA수평칙명현강저(P<0.05). 결론 Kallikrein가능통과감소조망인자bax、caspase-3적표체,억제뇌경사주변구역적신경세포조망,종이체도보호결혈신경세포,개선신경공능적작용.
Objective To investigate the effect of exogenous kallikrein on apoptosis of the neurons aroundthe cerebralinfarctareain rats. Methods Thirty rats wjth cerebral infarction induced by middle cerebral artery occlusion(MCAO)were assigned randomly into 3 groups(n=10),namely the blank control group,saline group,and pAdCMV-HTK group.In the pAdCMV-HTK group,kallikrein gene was delivered into the cerebral ischemie lesion via a replication-defective adenovims using stereotaetic injection technique, and the expression of exogenous kallikrein was detected immunohistoehemically.TUNEL staining was performed to evaluate the neuronal apoptosis around the infarct area,and RT-PCR used to detect the mRNA expressions ofbcl-2,bax and caspase-3 in the brain tissues. Results At 24 h aftertreatment there were some HTK expressed cells found in group C and peal(at 72 h after treatment.While compare with group B and group C,there existed significant difference(112±6.1,68±4.2,59±3.9,P<0.05).At 72 h after treatment,the NSS of group C was significantly lower than that ofgruop B and A(6.70±0.16,8.13±0.16,7.93±0.20,P<0.05);7 days after the treatment,the difference was more significant(5.14±0.18,7.82±0.14,7.91±0.10,P<0.01).Apoptotic cells were mostly seen around the infarct area.The ratsinpAdCMV-HTK group showed significantly reduced number of cells positive for TUNEL staining as compared to those in the saline and blank control groups at 3 days(10.1±0.9,16.7±1.1,and 20.4±0.8,respectively)and 7 days after the treatment(15.2±1.2,33.6±1.3,and 28.8±1.7,respectively)(P<0.05).The mRNA levels ofbc1-2.bax and caspasc-3 were elevated in all the groups at 24 h,peaked at 72 h,and decreased gradually till 7 days alter the treatment.Compared with those in the other two groups，bcl-2 mRNA level in the pAdCMV-HTK group increased slightly P>0.05) while bax and caspase-3 mRNA levels decreased markedly(P<0.05) 72 h and 7 days after the treatment.Conclusion Kallikrein can inhibit neuronal apoptosis around the cerebral infarct and improve the neurological fimction of rats following cerebral infarction probably by reducing the expressions of such apoptotic factors as bax and caspase-3.