Abstract:

BACKGROUND: Previous studies have proved that Rho-associated coiled-coil protein kinase (ROCK) 1 and ROCK2 involve in the process of apoptosis. On that basis, a hypothesis has been proposed that myocardial apoptosis can be inhibited by down-regulating the expression of ROCK1 and ROCK2.
OBJECTIVE: To verify whether the transfection of ROCK1-short hairpin RNA (ShRNA) and ROCK2-shRNA can effectively silence the expressions of ROCK1 and ROCK2 in rat cardiomyocytes and to screen for the shRNA with the best silencing effeciency.
METHODS: Three specific recombinant plasmids of ROCK1-shRNA and ROCK2-shRNA were constructed and identified respectively. Silencing effects of ROCK1 and ROCK2 in SD rat myocardial cells were assessed after shRNA transfection. And the shRNA with the best silencing efficiency was selected among these shRNAs.
RESULTS AND CONCLUSIONS: Western blot results showed that, compared with the control groups, the transfection of all three recombinant plasmids of ROCK1-shRNAs and ROCK2-shRNAs inhibited the expressions of ROCK1 and ROCK2 significantly (P < 0.05 or P < 0.01). Among these shRNAs, ROCK1-shRNA1 and ROCK2-shRNA2 showed the best silencing efficiency of (82.15±7.53)% and (89.42±5.81)%, respectively (P < 0.05). These findings show that the expressions of ROCK1 and ROCK2 in rat cardiomyocytes can be effectively inhibited by the transfection of ROCK1-shRNA and ROCK2-shRNA. And the shRNA with the best silencing efficiency was selected among these shRNAs.