CAJ | 학술논문

目的研究舒芬太尼后处理对心肌缺血/再灌注损伤(ischemiia/reperfusion injury,I/RI)细胞凋亡的影响以及与信号通路JAK2-STAT3的关系.方法健康杂种家犬24只,体重10 kg～15 kg,按随机数字表法分为4组(每组6只):假手术组(Sham组,只穿线,不结扎),心肌缺血/再灌注(ischemia/reperfusion,I/R)组,舒芬太尼后处理组(SPO组,于再灌注前5min静脉注射舒芬太尼0.6 μg/kg),舒芬太尼后处理+AG490组(SPO+AG490组,于再灌注前5 min静脉注射l mg/kg AG490,特异性的JAK2抑制剂),除Sham组外,所有犬心脏都经历30 min缺血和120 min再灌注.再灌注120 min时,取各组缺血区心肌组织,采用末端脱氧核苷酸转移酶介导的dUTP原位切口末端标记(terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling,TUNEL)法测定心肌细胞的凋亡情况,免疫组化法测定各组Bcl-2、Bax以及磷酸化STAT3 (p-ATAT3)蛋白的表达,并计算Bcl-2和Bax表达的比值(Bcl-2/Bax).结果再灌注120 min时,可在I/R组缺血区心肌组织中检测到大量凋亡心肌细胞(63.9±4.0)％,而舒芬太尼后处理显著降低心肌细胞凋亡指数(30.7±1.5)％；与Sham组比较,I/R组、SPO组和SPO+AG490组Bcl-2与Bax表达上调,I/R组Bcl-2/Bax比值降低,SPO组Bcl-2/Bax比值升高；舒芬太尼后处理使p-STAT3表达明显增加,特异性的阻断剂AG490抑制了舒芬太尼后处理对心肌I/RI凋亡的作用,即抑制p-STAT3表达的增加. 结论舒芬太尼后处理对心肌I/RI细胞凋亡有一定的抑制作用,通过激活JAK2-STAT3信号转导通路上调Bcl-2蛋白和下调Bax蛋白来发挥作用.
목적 연구서분태니후처리대심기결혈/재관주손상(ischemiia/reperfusion injury,I/RI)세포조망적영향이급여신호통로JAK2-STAT3적관계.방법 건강잡충가견24지,체중10 kg～15 kg,안수궤수자표법분위4조(매조6지):가수술조(Sham조,지천선,불결찰),심기결혈/재관주(ischemia/reperfusion,I/R)조,서분태니후처리조(SPO조,우재관주전5min정맥주사서분태니0.6 μg/kg),서분태니후처리+AG490조(SPO+AG490조,우재관주전5 min정맥주사l mg/kg AG490,특이성적JAK2억제제),제Sham조외,소유견심장도경력30 min결혈화120 min재관주.재관주120 min시,취각조결혈구심기조직,채용말단탈양핵감산전이매개도적dUTP원위절구말단표기(terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling,TUNEL)법측정심기세포적조망정황,면역조화법측정각조Bcl-2、Bax이급린산화STAT3 (p-ATAT3)단백적표체,병계산Bcl-2화Bax표체적비치(Bcl-2/Bax).결과 재관주120 min시,가재I/R조결혈구심기조직중검측도대량조망심기세포(63.9±4.0)％,이서분태니후처리현저강저심기세포조망지수(30.7±1.5)％；여Sham조비교,I/R조、SPO조화SPO+AG490조Bcl-2여Bax표체상조,I/R조Bcl-2/Bax비치강저,SPO조Bcl-2/Bax비치승고；서분태니후처리사p-STAT3표체명현증가,특이성적조단제AG490억제료서분태니후처리대심기I/RI조망적작용,즉억제p-STAT3표체적증가. 결론 서분태니후처리대심기I/RI세포조망유일정적억제작용,통과격활JAK2-STAT3신호전도통로상조Bcl-2단백화하조Bax단백래발휘작용.
Objective To investigate the anti-apoptotic effects of sufentanil postconditioning on myocardial ischemia/reperfusion injury (I/RI) and its relationship to the JAK2-STAT3 signaling pathway. Methods Twenty-four dogs were randomly divided into 4 groups:sham-operation group(group Sham),ischemia/reperfusion group(group I/R),sufentanil postconditioning+I/R group(group SPO),AG490+sufentanil postconditioning+I/R group(group SPO+AG490).Except sham group,all dogs subjected to 30 min of myocardial ischemia followed by 120 min of reperfusion.After reperfusion 2 h,the presence of apoptosis was determined quantitively by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL) methods,immunohistochemistry was used to detect the Bcl-2,Bax,p-STAT3 and protein of myocardial tissue. Results A significant number of TUNEL positive cells [ (63.9±4.0)％ ] were observed in myocardial tissue from hearts subjected to 30 min of myocardial ischemia followed by 120 min of reperfusion.Administration of sufentanil exerted a significant anti-apoptotic effect that has been conformed by reduced TUNEL-positive staining [ (30.7±1.5)％ ].Compared with the group Sham,expression of Bcl-2 and Bax is increased in myocardial group I/R,group SPO and group SPO+AG490.Bcl-2/Bax ratio is lower in group I/R and higher in group SPO; P-STAT3 activity was increased in the myocardial tissue after sufentanil postconditioning compared with that in group Sham (P＜0.05). Conclusions Sufentanil postconditioning provided myocardial protection to I/RI on dogs,the mechanism of myocardial protection is related to the inhibition of cell apoptosis via up-regulation of Bcl-2 expression and down-regulation Bax expression.