Influence of hydrogen sulfide on zymogen activation of homocysteine-induced matrix metalloproteinase-2 in H9C2 cardiocytes.

Abstract

To observe the influence of different concentrations of homocysteine (Hcy) and hydrogen sulfide (H2S) on the secretion and activation of matrix metalloproteinase-2 (MMP-2) in cardiocytes so as to search for new ways to fight against myocardial tissue fibrosis. Cardiocytes H9C2 was cultured invitro and different concentrations of Hcy and H2S were added for 6-h and 24-h cultivation. MTT cell proliferation assay was applied to test the activation change of cardiocytes H9C2 after affecting by different concentrations of Hcy and H2S. ELISA and MTT were employed to detect the expression and enzymatic activity of MMP-2. The H9C2 cell inhibition of activity was more significant with 1000μmol/L of Hcy as compared with other concentrations (P<0.001). With 2.5-100.0μmol/L Hcy and 0.1, 1.0 and 10.0mmol/L H2S, the activity of H9C2 did not change significantly (P>0.05). Hcy with concentrations of 10, 50 and 100μmol/L could increase the quantity of MMP-2 secreted by cardiocytes H9C2, and the interaction strength was concentration-dependent (P<0.05). After interacting with 100μmol/L of Hcy for 6h, the zymogen activation effect of MMP-2 was stronger than that of the 2.5-25μmol/L group (P<0.05). After interacting with Hcy and H2S (1.0mmol/L) for 6h and 24h, the activation effect of MMP-2 was stronger than those interacted with 10, 25, 50 and 100μmol/L of Hcy (P<0.05). Hcy can increase the production of MMP-2 secreted by H9C2 cell and improve its zymogen activation. Besides, the interaction strength is concentration-dependent; while H2S can up-regulate the activation of MMP-2 and co-promote the activation of MMP-2 with Hcy as well.