Molecular cloning of human heat shock protein 27 and its protective effects on oxidative damage induced by H_2O_2 in rat cardiomyocte cell line H9c2

Objective To clone human cardiac heat shock protein 27 gene and to determine the effects of HSP27 on the oxidative stress in rat cardiomyocyte cell line H9c2. Methods Full length of HSP27 cDNA obtained by RT PCR was constructed into pCDNA3 1 + . The recombinant plasmid was transfected into rat cardiomyocyte cell line H9c2 and stable transfection cell line was selected by G418. Then we observed the effects of HSP27 over expression on the LDH release and apoptosis induced H 2 O 2 in H9c2. Results (1) pCDNA3 1 + /HSP27 provided a sound expression of HSP27 in both 293T and H9c2. (2)LDH releasing induced by 0, 100, 250, 500,1000 μmol/L H 2 O 2 in HSP27 overexpression group and wild type group were as follows: 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. (3)Apoptosis induced by 150 μmol/L H 2 O 2 in HSP27 over expression group and wild type group were (10 693±1 122)% vs (4 027±1 628)%( P 0 01). Conclusions The over expression of human HSP27 could inhibit oxidative damage significantly in H9c2.