EFFECT OF SINI DECOCTION ON THE EXPRESSION OF CAVEOLIN-1 AND ENOS IN EAHY926 CELL INJURED BY HOMOCYSTEINE. (8th October 2012) - Yong, Liu
Abstract : Objectives: To detect the effect of Sini Decoction on the expression of Caveolin-1 and eNOS in EAhy926 cell injured by homocysteine. Methods: Model of EAhy926 cell injured by homocysteine was made, the protection on the EAhy926 cell of Sini Decoction with different dosages and different durations were observed. The effection of Sini Decoction on the expression of protein of Caveolin-1 and eNOS in EAhy926 cell were observed by Western-blot, and effection of Sini Decoction on the expression of mRNA of Caveolin-1 and eNOS in EAhy926 cell were observed by fluorescent quantitation PCR. Results: After model of EAhy926 cell injured by homocysteine was made, we found that cultured with 0.5, 1.0, 2.0, 4.0, 8.0 µmol/l homocysteine, cells grew less than cultured with normal culture medium, and with the increase of homocysteine concentration, the number of attached cell grew downwards obviously, as culturing with homocysteine 4.0 µmol/l for 24 h did lower damage to cells and could induce effective cell injuring, it was made to be the model of injury. To detect the effection of Sini Decoction on EAhy926 cell injured by homocysteine, well growing EAhy926 cells were cultured in culture plate. 24 h later, cells were cultured with DMEM medium containing 2% fetal calf serum for 8 h to make cells hungry, then cultured with medium containing Sini Decoction 0, 0.25, 0.5, 1.0 g/ml respectively for 30 min, then cultured with medium containing homocysteine 4.0 µmol/l for 24 h. It was found that, compared with control group, attached cells in Sini Decoction groups grew better, and attached cells in Sini Decoction 1.0 g/ml plus homocysteine 4.0 µmol/l group grew best. Detected by western-blot, it was found that, compared with control group, there was no obvious change of protein of Caveolin-1 and eNOS in Sini Decoction 1.0 g/ml group, but in homocysteine 4.0 µmol/l medol group, expression of Caveolin-1 protein enhanced obviously, expression of eNOS protein weakened obviously, and in Sini Decoction groups, expression of Caveolin-1 protein weakened, and expression of eNOS protein enhanced, and in Sini Decoction 1.0 g/ml plus homocysteine 4.0 µmol/l group it was the most obvious p<0.05). Detected by fluorescent quantitation, it was found that, compared with control group, there was no obvious change of mRNA of Caveolin-1 and eNOS in Sini Decoction 1.0 g/ml group, but in homocysteine 4.0 µmol/l medol group, expression of Caveolin-1 mRNA enhanced obviously, expression of eNOS mRNA weakened obviously, and in Sini Decoction groups, expression of Caveolin-1 mRNA weakened, and expression of eNOS mRNA enhanced, and in Sini Decoction 1.0 g/ml Plus homocysteine 4.0 µmol/l group, it was the most obvious p<0.05). Conclusions: Homocysteine may injure EAhy926 cell by enhancing the expression of caveolin-1 then suppressing the expression of eNOS, while Sini Decoction may protect EAhy926 cell by suppressing the expression of caveolin-1 then enhancing the expression of eNOS.