小窝蛋白3在小鼠糖尿病心肌病中的作用:与内质网应激的关系  

Role of caveolin 3 in diabetic cardiomyopathy in mice:relationship with endoplasmic reticulum stress

作　　者：张岳甫 周璐 苏娃婷[1] 夏中元[1] 雷少青[1] Zhang Yuefu;Zhou Lu;Su Wating;Xia Zhongyuan;Lei Shaoqing(Department of Anesthesiology,Renmin Hospital of Wuhan University,Wuhan 430000,China)

机构地区：[1]武汉大学人民医院麻醉科,武汉430000

出　　处：《中华麻醉学杂志》2023年第4期463-467,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81772049)。

摘　　要：目的评价小窝蛋白3(Cav-3)在小鼠糖尿病心肌病中的作用及其与内质网应激的关系。方法在体实验:清洁级健康成年雄性野生型小鼠16只,体质量18~20 g,采用随机数字表法分为2组(n=8):对照组(Control组)和糖尿病心肌病组(DCM组),另取Cav-3 KO小鼠8只为Cav-3 KO+糖尿病心肌病组(Cav-3 KO+DCM组)。采用高脂饮食联合腹腔一次性注射链脲佐菌素100 mg/kg的方法制备小鼠2型糖尿病模型,8周时采用心脏超声测定小鼠左心射血分数(EF)、左室短轴缩短率(FS)、左室收缩末期容积(LVESD)、左室舒张末期容积(LVEDD)。随后处死小鼠取心脏,HE染色观察心肌组织形态学结果。离体实验:小鼠来源的HL-1心肌细胞,采用随机数字表法分为3组(n=6):正常糖培养组(NG组)、高糖组(HG组)和高糖+甲基-β-环糊精组(HG+β-CD组)。高糖模型采用专用培养基加入50%葡萄糖至终浓度达到30 mmol/L,持续培养36 h。采用LDH、CCK8试剂盒评估细胞损伤情况。采用Western blot法检测心肌组织与HL-1细胞内质网应激相关蛋白结合免疫球蛋白(BiP)、C/EBP同源蛋白(CHOP)及剪接型X-box结合蛋白1(XBP1-s)的表达。结果在体实验:与Control组比较,DCM组和Cav-3 KO+DCM组小鼠进食量、饮水量及心脏质量/体质量增加,EF和FS降低,LVESD和LVEDD升高,心肌BiP、CHOP和XBP1-s表达上调,Cav-3表达下调(P<0.05),病理学损伤加重;与DCM组比较,Cav-3 KO+DCM组EF和FS降低,LVESD和LVEDD升高,心肌BiP、CHOP与XBP1-s表达上调,Cav-3表达下调(P<0.05),病理学损伤加重。离体实验:与NG组比较,HG组和HG+β-CD组心肌细胞存活率下降,LDH活性升高,BiP、CHOP与XBP1-s表达上调,Cav-3表达下调(P<0.05);与HG组比较,HG+β-CD组心肌细胞存活率下降,LDH活性升高,BiP、CHOP和XBP1-s表达上调,Cav-3表达下调(P<0.05)。结论Cav-3表达下调会加重小鼠糖尿病心肌损伤,其机制与内质网应激过度激活有关。Objective To evaluate the role of caveolin 3(Cav-3)in diabetic cardiomyopathy and the relationship with endoplasmic reticulum stress in mice.Methods This experiment was performed in two parts.PartⅠin vivo experiment Sixteen clean-grade healthy adult male wild type mice weighing 18-20 g,were divided into 2 groups(n=8 each)using a random number table method:control group(Control group)and diabetic cardiomyopathy group(DCM group).Another 8 Cav-3 KO mice were selected and served as Cav-3 KO+diabetic cardiomyopathy group(Cav-3 KO+DCM group).Type 2 diabetic models were developed by high fat diet combined with intraperitoneal injection of streptozotocin(100 mg/kg).The left ventricular ejection fraction(EF),left ventricular short axis shortening rate(FS),left ventricular end-systolic diameter(LVESD)and left ventricular end-diastolic diameter(LVEDD)were measured by B ultrasound at 8 weeks.Then the mice were sacrificed,and the myocardial histomorphology was observed using HE staining.PartⅡin vitro experiment HL-1 cardiomyocytes were divided into 3 groups(n=6 each)using a random number table method:normal glucose group(NG group),high glucose group(HG group)and high glucose+methyl-β-cyclodextrin group(HG+β-CD group).The high glucose model was prepared by adding 50%glucose to a specialized culture medium until the final concentration reached 30 mmol/L,and HL-1 cardiomyocytes were continuously cultivated for 36 h.The cellular injury was assessed using LDH and CCK8 kits.The expression of endoplasmic reticulum stress-related proteins binding immunoglobulin protein(BiP),C/EBP-homologous protein(CHOP)and X-box binding protein 1(XBP1-s)in myocardial tissues and HL-1 cells was detected by Western blot.Results In vivo experiment Compared with Control group,the food intake,water intake,and heart mass/body mass were significantly increased,EF and FS were decreased,LVESD and LVEDD were increased,the expression of BiP,CHOP and XBP1-s was up-regulated,the expression of Cav-3 was down-regulated(P<0.05),and the pathological damage was

关 键 词：细胞质膜微囊蛋白 糖尿病心肌病 内质网应激 

分 类 号：R587.2[医药卫生—内分泌] R542.2[医药卫生—内科学]