Molecular cloning of human heat shock protein 27 and study of its protective effects on oxidative damage in rat cardiomyocte H9c2

摘要

Objective: To clone human cardiac heat shock protein 27(HSP27) gene and to determine the effects of HSP27 on the oxidative stress in rat cardiomyocyte cell line H9c2. Methods: Full length of HSP27 cDNA which got by RT-PCR was constructed into pCDNA3.1~+ . The recombinant was transfected into rat cardiomyocyte cell line H9c2 and the stable transfection cell line was selected by G418. Then we observe the effects of HSP27 over-expression on LDH release and apoptosis induced H_2O_2 in H9c2. Results: ①pCDNA3.1~+ /HSP27 provided a sound expression of HSP27 in both 293T and H9c2. ②LDH releasing induced by 0, 100,250,500, 1000 μmol/L H_2O_2 in HSP27 over-expression group and wild type group were 0.396 ± 0.017 vs. 0.390± 0.009 (p > 0.05), 0.437±0. 014 vs. 0.416±0.015 (P< 0.05), 0.471±0.018 vs. 0.417±0.009 (P< 0.001), 0.505±0.030 vs. 0.657 ± 0.022(P < 0.001), 0.547 ± 0.027 and 0.661 ± 0.011 (P < 0.001), respectively. ③Apoptosis induced by 150 μmol/L H_2O_2 in HSP27 over-expression group and wild type group were (10.693±1.122)% vs. (4.027± 1.628)%(P< 0.01). Conclusion: We cloned and constructed human cardiac HSP27 gene successfully, and over-expression of human HSP27 could inhibit oxidative damage significantly in H9c2.