ABSTRACT
[Abstract ] Objective Cardiac shock wave therapy (CSWT) can promote arteriogenesis in ischemic myocardia , but the mo-lecular mechanism remains unclear .The study aimed to explore the effect of CSWT on arteriogenesis in human cardiac microvascular endothelial cells ( HCMEC ) and the role of focal adhesion kinase (FAK) and Calcium-activated potassium channels (KCa) in the sig-nal conduction pathway of CSWT arteriogenesis . Methods HC-MEC cells cultured in vitro were randomly divided into control group , CSWT group , CSWT +T ( FAK inhibitor PF-573228 ) group and CSWT+F( SCa inhibitor iberiotoxin ) group.Each group received one CSWT(0.09 mJ/mm2, 200Times) 48 h after added stimulant.24 hours'conventional culture later , tests were made on the levels of endothelial nitric oxide synthase ( eNOS ) and vascular endothelial growth factor ( VEGF) mRNA as well as the changes of related protein expression . Results ①QPCR test showed that eNOS , VEGF mRNA expressions increased in CSWT group compared with control group (4.61 ±0.19 vs 3.99 ±0.17, P<0.05), while compared with CSWT group, eNOS, VEGF mRNA expressions in CSWT +T group were decreased (0.62 ±0.10 vs 0.40 ±0.02, P<0.05), eNOS, VEGF mRNA expressions in CSWT +F group were also decreased (0.53 ±0.02 vs 0.64 ±0.02, P<0.05), all the differ-ences were of statistical significance .②Western blot showed that eNOS , VEGF protein expressions increased in CSWT group compared with control group(0.63 ±0.02 vs 0.43 ±0.02, P<0.05), while compared with CSWT group , eNOS, VEGF protein expressions in CSWT+T group were decreased (0.36 ±0.01 vs 0.29 ±0.02, P<0.05), eNOS, VEGF protein expressions in CSWT +F group were also decreased (0.37 ±0.02 vs 0.30 ±0.02, P<0.05), all the differences were of statistical significance . Conclusion CSWT can improve eNOS , VEGF mRNA and protein expressions in HCMEC cells and FAK and KCa may participate in the signal conduction pathway of CSWT arteriogenesis .
ABSTRACT
[ ABSTRACT] AIM:To investigate the protective effect of naringin ( Nar) on the injury of human umbilical vein endothelial cells ( HUVECs) induced by 33 mmol/L high glucose ( HG) and to explore its possible mechanisms.METH-ODS:The injury model was established by treating HUVECs with HG medium for the indicated time (6, 12, 24, 48 and 72 h) , and then the levels of NO, eNOS and p-eNOS were detected, respectively.The effects of Nar on high glucose-in-duced endothelial cell injury were observed.HUVECs were treated with Nar at concentrations of 5, 10, 25, 50 and 100 mg/L for 6 h, 12 h, 24 h, 36 h and 48 h.The levels of NO in the supernatants were measured.The effects of Nar on HG-injured HUVECs were explored by treating the cells with 10 μmol/L of LY294002, a PI3K inhibitor, or 0.5 μmol/L of AKT inhibitorⅣ, an AKT inhibitor, and then the levels of NO, PI3K, AKT, eNOS and their phosphorylated proteins were determined by Western blot.RESULTS:Nar at concentration of 50 mg/L significantly attenuated the injury of endothelial cells induced by high glucose ( P<0.01) , and the protective effects of Nar were abolished by pretreating with the inhibitor of PI3K or AKT (P<0.01).CONCLUSION: Nar protects endothelial cells against the injury induced by high glucose through PI3K/AKT/eNOS pathway.