The effect of curcumin on DNA content, mitochondrial transmembrane potential and calcium of rabbit cultured retinal pigment epithelial cells

Abstract

To investigate the Change of DNA content (apoptosis rate), mitochondrial transmembrane potential (DeltaPsim) and calcium (Ca(2+)) of rabbit retinal pigment epithelial (RPE) cells cultured with curcumin. It was an experimental study. The RPE cells were dissociated from rabbit eyes and cultured. The RPE cells in the 4(th) passage were divided into 2 groups: curcumin group and control group (10% FBS-EMDM contains 0.05% dimethyl sulfoxide). The curcumin group contained 3 mass concentration: 10 mg/L, 15 mg/L and 20 mg/L. The MTT assay was used to evaluate the inhibition effect of RPE cells cultured with curcumin after 24 h, 48 h, 72 h and 96 h respectively. The IC(50) value in 24 h, 48 h, 72 h and 96 h were gotten by Linear Regression. Flow cytometry was performed to detect the change of DNA content (apoptosis rate), DeltaPsim and Ca(2+) of RPE cells cultured with curcumin (15 mg/L) after 8 h, 24 h, 48 h and 72 h respectively. RPE cells were significantly inhibited by curcumin in a dose dependent and time dependent manner. The IC(50) value of curcumin at 24 h, 48 h, 72 h and 96 h was 29.31 mg/L, 17.50 mg/L, 13.24 mg/L and 10.99 mg/L respectively. Ca(2+) was significantly increased at 8 h, 24 h, 48 h and 72 h after cultured with curcumin (15 mg/L) than that of the control group respectively (t = 7.50, 10.61, 20.74, 21.14, P < 0.01), and DeltaPsim was significantly decreased at 8 h, 24 h, 48 h and 72 h after cultured with curcumin (15 mg/L) than that of the control group respectively (t = 7.50, 11.74, 14.91, 15.29, P < 0.01). There was no change of DNA content in RPE cells at 8h after cultured with curcumin (15 mg/L), but significantly lower than that of the control group at 24 h, 48 h and 72 h respectively (t = 10.00, 14.68, 13.68, P < 0.01). The apoptosis of RPE cells induced by curcumin is caused by increase of Ca(2+) and decrease of DeltaPsim that causes decrease of DNA content. The RPE cells are significantly inhibited by curcumin, which may become a potential drug to prevent and treat proliferative vitreoretinopathy.