CAJ | 학술논문

目的观察急性全脑缺血再灌注大鼠脑组织Caspase-3表达及凋亡情况,探讨δ阿片受体激动剂DADLE对急性全脑缺血再灌注脑组织的影响.方法将50只健康雌性SD大鼠(180 ～220 g)随机分为五组:假手术组(Sham,n=10):只暴露血管而不夹闭；缺血再灌注组(I/R,n=10):采用改良的二血管阻断加低血压法建立急性全脑缺血再灌注模型,缺血15 min后给予再灌注；DADLE处理组分为DADLE 2 mg/kg组(A,n=10)、DADLE3 mg/kg组(B,n=10)、DADLE5 mg/kg组(C,n=10):在I/R组的基础上于再灌注前经颈外静脉注射DADLE.手术结束后取大脑予4％多聚甲醛固定,原位末端杂交法(Tunel染色)检测细胞凋亡及免疫组化法检测Caspase-3蛋白表达情况.结果 Tunel染色结果显示,I/R组与Sham组相比凋亡细胞明显增加(P＜0.05),DADLE处理组与I/R组相比,凋亡细胞有显著降低(P＜0.05),其中B组、C组凋亡细胞较A组减少更为显著(P＜0.05),B组、C组比较差异无统计学意义(P＞0.05).Sham组Caspase-3阳性细胞明显低于其他各组(P＜0.05),DADLE处理组比较I/R组比较,Caspase-3阳性细胞数明显减少(P＜0.05),其中B组、C组阳性细胞较A组减少更为显著(P＜0.05),B组、C组间差异无统计学意义(P＞0.05).结论在急性大鼠全脑缺血再灌注时,细胞凋亡参与了神经元的损伤,通过上调Caspase-3蛋白的表达介导凋亡的发生,而DADLE可以显著的抑制Caspase-3蛋白表达,从而减少细胞凋亡,发挥脑保护的作用.
목적 관찰급성전뇌결혈재관주대서뇌조직Caspase-3표체급조망정황,탐토δ아편수체격동제DADLE대급성전뇌결혈재관주뇌조직적영향.방법 장50지건강자성SD대서(180 ～220 g)수궤분위오조:가수술조(Sham,n=10):지폭로혈관이불협폐；결혈재관주조(I/R,n=10):채용개량적이혈관조단가저혈압법건립급성전뇌결혈재관주모형,결혈15 min후급여재관주；DADLE처리조분위DADLE 2 mg/kg조(A,n=10)、DADLE3 mg/kg조(B,n=10)、DADLE5 mg/kg조(C,n=10):재I/R조적기출상우재관주전경경외정맥주사DADLE.수술결속후취대뇌여4％다취갑철고정,원위말단잡교법(Tunel염색)검측세포조망급면역조화법검측Caspase-3단백표체정황.결과 Tunel염색결과현시,I/R조여Sham조상비조망세포명현증가(P＜0.05),DADLE처리조여I/R조상비,조망세포유현저강저(P＜0.05),기중B조、C조조망세포교A조감소경위현저(P＜0.05),B조、C조비교차이무통계학의의(P＞0.05).Sham조Caspase-3양성세포명현저우기타각조(P＜0.05),DADLE처리조비교I/R조비교,Caspase-3양성세포수명현감소(P＜0.05),기중B조、C조양성세포교A조감소경위현저(P＜0.05),B조、C조간차이무통계학의의(P＞0.05).결론 재급성대서전뇌결혈재관주시,세포조망삼여료신경원적손상,통과상조Caspase-3단백적표체개도조망적발생,이DADLE가이현저적억제Caspase-3단백표체,종이감소세포조망,발휘뇌보호적작용.
Objective To observe the expression and apoptosis of brain Caspase-3 of acute cerebral ischemia reperfusion rat,and to investigate delta opioid receptor agonist DADLE on acute cerebral ischemia reperfusion brain of rat.Methods Fifty healthy female SD rats (180-220 g) were randomly divided into 5 groups:sham operation group (Sham,n =10):only exposed blood vessels without clipping; ischemia reperfusion group (I/R,n =10):a modified two vessel occlusion with hypotension method to establish acute global cerebral ischemia reperfusion models,lack of blood 15 min given after reperfusion ; DADLE treatments were divided into DADLE 2 mg/kg group(A,n =10),DADLE 3 mg/kg group (B,n =10),DADLE 5 mg/kg group (C,n =10):in the I/R group on the basis of the prior to reperfusion via external jugular vein injection of DADLE.After the operation,from the brain to the fixed in 4％ paraformaldehyde,in situ hybridization (Tunel stain) detection of cell apoptosis and immunohistochemistry assay for detection of Caspase-3 protein expression.Results Tunel staining showed that I/R group compared with Sham group,the apoptosis cells were significantly increased (P ＜ 0.05),DADLE group than in I/R group compared,apoptotic cells decreased significantly (P ＜ 0.05),B group,C group of apoptotic cells was lower than A group was more significant (P ＜ 0.05),B group,C group showed no significant difference (P ＞ 0.05).Group Sham Caspase-3 positive cells were significantly lower than the other groups (P ＜ 0.05),DADLE group than in I/R group,the number of Caspase-3positive cells was significantly reduced (P ＜0.05),positive cells of are group B and C was lower than group A (P ＜ 0.05),group B and C had no significant difference (P ＞ 0.05).Conclusions During acute cerebral ischemia reperfusion in rats,apoptosis is involved in neuronal damage,through the up-regulation of expression of Caspase-3 protein mediated apoptosis,while DADLE can sigrnificantly inhibit the expression of Caspase-3 protein,thereby reducing the cell apoptosis,cerebral protection role play.