HK2和VDAC1在二乙酰吗啡致心肌细胞凋亡中的作用  

Role of HK2 and VDAC1 in Diacetylmorphine-induced Cardiomyocyte Apoptosis

作　　者：肖锦玲 管雅玲 朱森森 庄梦婕 苏丽萍[2] 蒲红伟[3] Jinling XIAO;Yaling GUAN;Sensen ZHU;Mengjie ZHUANG;Liping SU;Hongwei PU(Dept.of Pathology,School of Basic Medical Sciences,Xinjiang Medical University,Urumqi,Xinjiang Uygur Autonomous Region Xinjiang 830011;Dept.of Pathology,The 1st Affiliated Hospital of Xinjiang Medical University,Urumqi Xinjiang Uygur Autonomous Region 830011;Dept.of Discipline Construction,Urumqi Xinjiang Uygur Autonomous Region 830011,China)

机构地区：[1]新疆医科大学基础医学院病理学教研室,新疆维吾尔自治区乌鲁木齐830011 [2]新疆医科大学第一附属医院病理科,新疆维吾尔自治区乌鲁木齐830011 [3]新疆医科大学第一附属医院学科建设科,新疆维吾尔自治区乌鲁木齐830011

出　　处：《昆明医科大学学报》2024年第2期7-13,共7页Journal of Kunming Medical University

基　　金：国家自然科学基金资助项目(8216020119);新疆维吾尔自治区研究生科研创新项目(XJ2023G192)。

摘　　要：目的探讨HK2和VDAC1在二乙酰吗啡致心肌细胞凋亡中的作用。方法采用剂量递增方法,建立二乙酰吗啡成瘾大鼠模型。将40只SD大鼠随机分为3组,正常组(Control,n=10)皮下注射等量生理盐水;模型组(Model,n=15)首次皮下注射二乙酰吗啡剂量为5 mg/kg,以后逐日递增剂量2.5 mg/(kg·d),连续注射20 d;模型+10 D组(Model+10D,n=15)在模型组基础上继续递增剂量至第10天。采用ELISA法检测乳酸脱氢酶(lactate dehydrogenase,LDH)、谷草转氨酶(glutamic oxaloacetic transaminase,GOT)含量;采用HE染色观察各组心肌组织病理变化;采用TUNEL染色检测各组心肌组织细胞凋亡;采用免疫组化、RT-qPCR和Western blot检测HK2、VDAC1及凋亡相关因子的mRNA和蛋白的表达变化。结果HE染色发现随着二乙酰吗啡干预时间的延长,心肌组织表现出不同程度的损伤。与正常组相比,模型组中血清LDH、GOT含量及心肌凋亡率升高,HK2和抗凋亡因子Bcl-2的mRNA和蛋白水平下降,VDAC1和促凋亡因子Bax、Caspase-3的mRNA和蛋白水平升高,Clevead Caspase-3蛋白水平升高;与正常组相比,模型+10 D组中以上指标,差异有统计学意义(P<0.05)。结论二乙酰吗啡可导致心肌细胞凋亡,HK2和VDAC1可能参与了二乙酰吗啡致心肌细胞凋亡的过程。Objective To investigate the role of HK2 and VDAC1 in diacetylmorphine-induced cardiomyocyte apoptosis.Methods A dose-escalation method was used to establish a rat model of diacetylmorphine addiction.Forty SD rats were randomly divided into three groups,the normal group(n=10)was injected with an equal amount of saline subcutaneously,the model group(n=15)was injected with 5 mg/kg of diacetylmorphine for the first time,and then the dose was increased by 2.5 mg/(kg·d)day by day for 20 days,and the group of model+10 D(n=15)continued to increase the dose based on the model group up to the 10th day.Lactate dehydrogenase(LDH)and glutamic oxaloacetic transaminase(GOT)were detected by ELISA;HE staining was used to observe the pathological changes of myocardial tissues in each group;TUNEL staining was used to detect apoptosis in myocardial tissues in each group;and immunohistochemistry,RT-q-analysis,and immunochemistry were used to detect apoptosis in myocardial tissues in each group.Immunohistochemistry,RT-qPCR and Western bl-ot were used to detect the mRNA and protein expression of HK2,VDAC1 and apoptosis-related factors.Results HE staining revealed that myocardial tissues exhibited different degrees of damage with the prolongation of diacetylmorphine intervention.Compared with the normal group,serum LDH,GOT content and myocardial apoptosis rate increased in the model group,mRNA and protein levels of HK2 and anti-apoptotic factor Bcl-2 decreased,mRNA and protein levels of VDAC1 and pro-apoptotic factors Bax and Caspase-3 increased,and the protein level of Clevead Caspase-3 increased;in the model+10 D group the above indexes,there was a statistically significant difference(P<0.05).Conclusion Diacetylmorphine can cause cardiomyocyte apoptosis,and VDAC1 may be involved in the process of cardiomyocyte apoptosis caused by diacetylmorphine.

关 键 词：二乙酰吗啡 心肌凋亡 己糖激酶2 电压依赖性阴离子通道1 裂解半胱氨酸天冬氨酸蛋白酶-3 

分 类 号：R966[医药卫生—药理学]