CAJ | 학술논문

目的评价磷脂酰肌醇3-激酶/蛋白激酶B(PI3 K/Akt)信号通路在氢气抑制大鼠心肌缺血再灌注时细胞凋亡中的作用.方法健康雄性SD大鼠40只,体重300 ～ 350 g,采用随机数字表法分为4组(n=10):假手术组(S组)、心肌缺血再灌注组(I/R组)、氢气组(H组)和氢气+PI3K特异性阻断剂LY294002组(HL组).采用结扎左冠状动脉前降支30 min,再灌注120 min的方法制备心肌缺血再灌注损伤模型.H组和HL组分别于再灌注即刻腹腔注射浓度为99.6％的氢气5 ml/kg,HL组于氢气注射前鼠尾静脉注射LY294002 0.3 mg/kg.于再灌注120 min时取右颈总动脉血样2 ml,测定血清肌酸激酶同工酶(CK-MB)和乳酸脱氢酶(LDH)活性;采用TUNEL染色法检测心肌细胞凋亡情况,并计算凋亡指数(AI),免疫组化法检测Bcl-2、Bax及caspase-3的表达.计算Bcl-2/Bax比值.结果与S组比较,I/R组血清CK-MB、LDH活性及AI均升高,心肌Bcl-2、Bax、caspase-3表达上调,Bcl-2/Bax比值降低(P＜ 0.01);与I/R组比较,H组血清CK-MB、LDH活性及AI均降低,心肌Bcl-2表达上调,Bax和caspase-3表达下调,Bcl-2/Bax比值升高(P＜0.01),HL组上述指标差异无统计学意义(P＞0.05);与H组比较,HL组血清CK-MB、LDH活性及AI升高,心肌Bcl-2表达下调,Bax和caspase-3表达上调,Bcl-2/Bax比值降低(P＜0.01).结论氢气可通过激活PI3K/Akt信号通路,进而上调抗凋亡蛋白Bcl-2表达及下调促凋亡蛋白Bax和caspase-3的表达,抑制大鼠心肌缺血再灌注时细胞凋亡.
목적 평개린지선기순3-격매/단백격매B(PI3 K/Akt)신호통로재경기억제대서심기결혈재관주시세포조망중적작용.방법 건강웅성SD대서40지,체중300 ～ 350 g,채용수궤수자표법분위4조(n=10):가수술조(S조)、심기결혈재관주조(I/R조)、경기조(H조)화경기+PI3K특이성조단제LY294002조(HL조).채용결찰좌관상동맥전강지30 min,재관주120 min적방법제비심기결혈재관주손상모형.H조화HL조분별우재관주즉각복강주사농도위99.6％적경기5 ml/kg,HL조우경기주사전서미정맥주사LY294002 0.3 mg/kg.우재관주120 min시취우경총동맥혈양2 ml,측정혈청기산격매동공매(CK-MB)화유산탈경매(LDH)활성;채용TUNEL염색법검측심기세포조망정황,병계산조망지수(AI),면역조화법검측Bcl-2、Bax급caspase-3적표체.계산Bcl-2/Bax비치.결과 여S조비교,I/R조혈청CK-MB、LDH활성급AI균승고,심기Bcl-2、Bax、caspase-3표체상조,Bcl-2/Bax비치강저(P＜ 0.01);여I/R조비교,H조혈청CK-MB、LDH활성급AI균강저,심기Bcl-2표체상조,Bax화caspase-3표체하조,Bcl-2/Bax비치승고(P＜0.01),HL조상술지표차이무통계학의의(P＞0.05);여H조비교,HL조혈청CK-MB、LDH활성급AI승고,심기Bcl-2표체하조,Bax화caspase-3표체상조,Bcl-2/Bax비치강저(P＜0.01).결론 경기가통과격활PI3K/Akt신호통로,진이상조항조망단백Bcl-2표체급하조촉조망단백Bax화caspase-3적표체,억제대서심기결혈재관주시세포조망.
Objective To evaluate the role of phosphoinositide 3 kinase/protein kinase B (PI3K/Akt) signaling pathway in hydrogen-induced inhibition of cell apoptosis during myocardial ischemia/reperfusion (I/R) in rats.Methods Forty healthy male Sprague-Dawley rats,weighing 300-350 g,were randomly allocated into 4 groups (n =10 each) using a random number table:sham operation group (S group),I/R group,hydrogen group (group H),and hydrogen + LY294002 group (group HL).Myocardial I/R was induced by occlusion of the anterior descending branch of left coronary artery for 30 min followed by 120 min reperfusion.In H and HL groups,99.6 ％ hydrogen 5 ml/kg was injected intraperitoneally immediately after beginning of reperfusion,and in addition LY294002 (0.3 mg/kg) was injected through the caudal vein before hydrogen injection in group HL.Arterial blood samples were collected at the end of 120 min reperfusion for determination of serum creatine kinase isoenzyme-MB (CK-MB) and lactate dehydrogenase (LDH) activities.The rats were then sacrificed.Myocardial apoptosis was detected by TUNEL and apoptosis index (AI) was calculated.The expression of Bcl-2,Bax and caspase-3 was detected by immuno-histochemistry.Bcl-2/Bax ratio was calculated.Results The serum CK-MB and LDH activities and AI were significantly increased,the expression of myocardial Bcl-2,Bax and caspase-3 was upregulated,and the ratio of Bcl-2/Bax was decreased in group I/R as compared with group S.Compared with group I/R,the serum CK-MB and LDH activities and AI were significantly decreased,the expression of myocardial Bcl2 was up-regulated,while the expression of myocardial Bax and caspase-3 was down-regulated,and the ratio of Bcl-2/Bax was increased in group H,and no significant changes were found in the parameters mentioned above in group HL.The serum CK-MB and LDH activities and AI were significantly increased,the expression of myocardial Bcl-2 was down-regulated,while the expression of myocardial Bax and caspase-3 was up-regulated,and Bcl-2/Bax ratio was decreased in group HL as compared with group H.Conclusion Hydrogen can activate the PI3K/Akt signaling pathway,and further up-regulates Bcl-2 expression and down-regulates Bax and Caspase-3 expression,thus inhibiting cell apoptosis during myocardial I/R in rats.