ABSTRACT
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. Part Ⅰ in 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. Part Ⅱ in 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 aggravated in DCM group and Cav-3 KO+ DCM group. Compared with DCM group, EF and FS were significantly 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 aggravated in Cav-3 KO+ DCM group. In vitro experiment Compared with NG group, the cell viability was significantly decreased, LDH activity was increased, the expression of BiP, CHOP and XBP1-s was up-regulated, and the expression of Cav-3 was down-regulated in HG group and HG+ β-CD group ( P<0.05). Compared with HG group, the cell viability was significantly decreased, LDH was increased, the expression of BiP, CHOP and XBP1-s was up-regulated, and the expression of Cav-3 was down-regulated in HG+ β-CD group ( P<0.05). Conclusions:Down-regulation of Cav-3 expression aggravates myocardial injury in diabetes mellitus, and the mechanism is related to excessive activation of endoplasmic reticulum stress in mice.
ABSTRACT
Objective To evaluate the effects of N-acetycysteine on the expression of caveolin-3 (Cav-3) during myocardial ischemia-reperfusion (I/R) in diabetic rats.Methods Twenty-four pathogenfree healthy adult male Sprague-Dawley rats,weighing 230-270 g,were divided into 3 groups (n =8 each) using a random number table:myocardial I/R group (group I/R),diabetes mellitus plus myocardial I/R group (group D) and N-acetycysteine group (group NAC).Diabetes mellitus was induced by injection of streptozotocin 60 mg/kg via the tail vein and confirmed by blood glucose ≥ 16.7 mmol/L 3 days later.At 1 week after successful establishment of the model,N-acetycysteine 1.5 g · kg-1 · d-1 was injected through a gastric tube into stomach for 4 consecutive weeks in group NAC,and the equal volume of normal saline was given for 4 consecutive weeks in I/R and D groups.Myocardial I/R was then induced by 30 min ligation of the left anterior descending branch of the coronary artery followed by 2 h of reperfusion.At the end of reperfusion,the myocardial infarct size was determined by triphenyl tetrazolium chloride staining,the levels of serum creatine kinase-MB (CK-MB) and 15-F2t-Isoprostane were measured by enzyme-linked immunosorbent assay,and the expression of myocardial Cav-3,Akt,phosphor-Akt (p-Akt),endothelial nitric oxide synthase (eNOS) and phosphor-eNOS (p-eNOS) was detected by Western blot.Results Compared with group I/R,the myocardial infarct size and levels of serum CK-MB and 15-F2t-Isoprostane were significantly increased,and the myocardial Cav-3,p-Akt and p-eNOS expression and NO level were decreased in group D (P＜0.05).Compared with group D,the myocardial infarct size and levels of serum CK-MB and 15-F2t-Isoprostane were significantly decreased,and the myocardial Car-3,p-Akt and p-eNOS expression and NO level were increased in group NAC (P＜0.05).Conclusion N-acetycystein can activate Akt/eNOS/NO signaling pathway through up-regulating myocardial Cav-3 expression,thus reducing myocardial I/R injury in diabetic rats.