BACKGROUND:Brahma-related gene 1 (Brg1), a catalytic subunit of an important chromatin remodeling complex, has been considered as a key nuclear transcriptional factor, and tends to be decreased in diabetic cardiomyopathy.
OBJECTIVE:To construct an adenovirus vector carrying Brg1, and observe its protective role in oxidative stress induced-cardiomyocyte apoptosis.
METHODS:The recombinant adenovirus plasmid was linearized and transfected into HEK293 cel s using Fugene HD for packaging and amplification. The adenovirus particles were further purified, quantified, and sequential y transfected to cardiomyocytes of neonatal Sprague-Dawley rats. The Adeno-EGFP transfected and non-transfected cardiomyocytes were used as control group. 24 hours later, the transfection efficiency was observed by fluorescent microscope, and expressions of Brg1 mRNA and protein were detected by quantified PCR and western blotting. After treatment with 100 μmol/L H2O2 for 12 hours, the expressions of Brg1 protein and cleaved-Caspase 3 were measured by western blotting, and cel apoptosis was analyzed by flow cytometry.
RESULTS AND CONCLUSION:(1) The recombinant adenovirus vector of Brg1 had been successful y transfected into cardiomyocytes with higher expressions of Brg1 mRNA and protein, and the transfection efficiency reached more than 90%. (2) After H2O2 treatment, the Brg1 was significantly down-regulated in contrast to the up-regulation of cleaved-Caspase 3;the flow cytometry data showed that the apoptotic cel s were increased. But in Adeno-Brg1 transfected cardiomyocytes, the H2O2 induced cel apoptosis was significantly decreased compared with non-transfected cel s and empty vector transfected cel s. (3) These results suggest that oxidative stress can directly inhibit the Brg1 expression, and overexpression of Brg1 can protect the cardiomyocytes from cel apoptosis induced by oxidative stress.

Aim Carvacrol ( CAR ) , possesses a wide variety of pharmacological properties including antioxi-dant and anti-inflammatory potential. The present stud-y is designed to investigate the effect of CAR on glu-cose and lipid metabolism in type 1 diabetic mice. Methods Diabetes was induced by intraperitoneal( i. p) injection of streptozotocin into male mice at the dose of 45 mg·kg-1 body weight( BW) . Mice were divided into three different groups containing eight to twelve in each. Age matched male C57 mice were used as nor-mal controls. Group I diabetes, Group Ⅱ and Ⅱ in-jected with CAR at 10 and 20 mg · kg-1 BW respec-tively once daily. After CAR injection 2, 4 or 6 weeks, the rats were weighted and the plasma concen-trations of glucose, total cholesterol( TC) , triglycerides (TG), Glutamic oxalacetic transaminase(AST), Ala-nine transaminase( ALT) levels were enzymatically de-termined using commercial kits. Results STZ-induced C57 BL/6 J diabetic mice showed an elevation in serum glucose, TG, ALT, AST and LDH levels. Compared to diabetic mice, administration of CAR resulted in sig-nificant decreases(P <0. 05) in plasma glucose, TG and LDH levels in a dose dependent manner, but no effect on elevated TC, ALT and AST levels. Conclu-sion These major findings provide evidence that CAR has anti diabetic property and it has the potential for development into a drug to prevent hyperglycemia, re-duce blood lipids and protect the dammaged organs.