乌司他丁对出血坏死性胰腺炎兔心肌细胞凋亡和Bcl-2、Bax蛋白表达的影响

目的探讨乌司他丁对出血坏死性胰腺炎兔心肌细胞凋亡和Bcl-2、Bax蛋白表达的影响。方法 24只新西兰大白兔分为对照组、1万U.kg-1乌司他丁组、2万U.kg-1乌司他丁组,每组8只。逆行胰管注射质量分数为5%牛磺胆酸钠溶液制作急性出血坏死性胰腺炎(ANP)模型。所有兔于模型制备成功结束后稳定30 min,阻断左冠状动脉前降支60 min,再灌注180 min。在阻断左冠状动脉前降支近端前对照组经静脉注射生理盐水2 mL.kg-1,1万U.kg-1乌司他丁组经静脉注射乌司他丁1万U.kg-1;2万U.kg-1乌司他丁组经静脉注射乌司他丁2万U.kg-1。于灌注结束后立即将兔处死,末端脱核苷酸转移酶介导的dUTP缺口标记技术观察兔缺血再灌注心肌细胞凋亡的变化,免疫组织化学法检测凋亡相关调控基因Bcl-2、Bax蛋白的表达。结果 1万U.kg-1乌司他丁组、2万U.kg-1乌司他丁组心肌细胞凋亡率均低于对照组(P<0.05),2万U.kg-1乌司他丁组心肌细胞凋亡率低于1万U.kg-1乌司他丁组(P<0.05)。1万U.kg-1乌司他丁组、2万U.kg-1乌司他丁组Bcl-2表达均显著高于对照组(P<0.05),2万U.kg-1乌司他丁组Bcl-2蛋白表达高于1万U.kg-1乌司他丁组(P<0.05);1万U.kg-1乌司他丁组、2万U.kg-1乌司他丁组Bax蛋白表达均低于对照组(P<0.05),2万U.kg-1乌司他丁组Bax蛋白表达低于1万U.kg-1乌司他丁组,但差别无统计学意义(P>0.05)。结论乌司他丁能明显减轻ANP兔心肌细胞凋亡,其抑制心肌细胞凋亡的机制可能是通过调节凋亡相关调控基因Bcl-2及Bax的不同表达来发挥作用。

Effect of ulinastain on the myocardial cell apoptosis and the expression of Bcl-2 and Bax protein in rabbits model with acute hemorrhagic necrotizing pancreatitis

Objective To investigate the effect of ulinastain on the myocardial cell apoptosis and the expression of Bcl-2 and Bax protein in rabbits with acute hemorrhagic necrotizing pancreatitis（ANP）.Methods Twenty-four rabbits were divided into control group,10 000 U·kg-1 ulinastain group and 20 000 U·kg-1 ulinastain group,eight rabbits in each group.The models of ANP was established by retrograde injection of 5% sodium taurocholate solution into the biliopancreatic duct in the rabbits.After the models were established successfully,all the rabbits were stabilized for 60 min,the left anterior descending coronary artery was blocked for 60 min and reperfusion for 180 min.The control group,10 000 U·kg-1 ulinastain group and 20 000 U·kg-1 ulinastain group were given saline solution,10 000 U·kg-1 ulinastain and 20 000 U·kg-1 ulinastain before blocking the left anterior descending coronary artery,respectively.The rabbits were sacrificed after reperfusion.The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling was used to test the myocardial cell apoptosis.The expressions of Bcl-2 and Bax protein were detected by immunohistochemical method.Results The myocardial cell apoptosis rate in 10 000 U·kg-1 ulinastain group and 20 000 U·kg-1 ulinastain group was lower than that in control group（P0.01）,and the myocardial cell apoptosis rate in 20 000 U·kg-1 ulinastain group was lower than that in 10 000 U·kg-1 ulinastain group（P0.05）.The expression of Bcl-2 in 10 000 U·kg-1 ulinastain group and 20 000 U·kg-1 ulinastain group was higher than that in control group（P0.01）,and the expression of Bcl-2 in 20 000 U·kg-1 ulinastain group was higher than that in 10 000 U·kg-1 ulinastain group（P0.05）.The expressions of Bax in 10 000 U·kg-1 ulinastain group and 20 000 U·kg-1 ulinastain group were lower than those in control group（P0.05,P0.01）,and the expression of Bax in 20 000 U·kg-1 ulinastain group was lower than that in 10 000 U·kg-1 ulinastain group,but the difference was no statistically significant（P0.05）.Conclusion Ulinastain can relieve the myocardial cell apoptosis in rabbits with ANP.The inhibition of myocardial cell apoptosis mechanisms could be related to the reduction the expression of Bax and increasing the expression of Bcl-2.