Influence of puerarin on proliferative activity of human fetal scleral fibroblasts and expression of collagen type I and matrix metalloproteinase-2 in power frequency electromagnetic field

Abstract

To investigate the influence of puerarin on the proliferative activity of human fetal scleral fibroblasts (HFSFs) and the expression of collagen type I and matrix metalloproteinase-2 (MMP-2) in HFSFs in the power frequency electromagnetic field. HFSFs were cultured and divided into four radiation groups (0.2 mT, 50 Hz) and control group. Puerarin at concentrations of 0.0, 1.0, 5.0, and 10.0 μmol/L was added to the four radiation groups, respectively. The methyl thiazolyl tetrazolium colorimetry was used to measure the influence on the proliferative activity of HFSFs, Western blot and RT-qPCR were used to measure the protein and mRNA expression of collagen type I and MMP-2 induced by puerarin in the power frequency electromagnetic field. The radiation groups and control group showed significant increases in the proliferative activity of HFSFs over the culture time (F= 959.472 and 279.468, both P<0.01). At 24 and 48 hours, the radiation groups showed significantly lower proliferative activity than the control group (0.432±0.038/0.591±0.017 vs 0.536±0.034/0.801±0.020, both P<0.01). With the increasing concentration of puerarin (0.0, 1.0, 5.0, and 10.0 μmol/L) , the proliferative activity of HFSFs significantly increased, with A values of 0.598±0.031, 0.809±0.041, 0.910±0.037, and 0.983±0.054, respectively (P<0.05). After exposure for 24 hours, the radiation groups showed significantly lower protein expression of collagen type I and significantly higher protein expression of MMP-2 (t=7.917 and 7.831, both P<0.01) ; compared with the 0.0 μmol/L puerarin group, the 1.0, 5.0, and 10.0 μmol/L puerarin groups showed significant increases in the protein expression of collagen type I and significant reductions in the protein expression of MMP-2 (all P<0.01). Compared with the control group, the radiation groups showed significant reductions in the mRNA expression of collagen type I and MMP-2 (t=17.293 and 16.378, both P<0.01) ; compared with the 0.0 μmol/L puerarin group, the 1.0, 5.0, and 10.0 μmol/L puerarin groups showed significant increases in the mRNA expression of collagen type I and significant reductions in the mRNA expression of MMP-2 (P<0.01). Puerarin can inhibit the reduction in the proliferative activity of HFSFs in the power frequency electromagnetic field, upregulate the expression of collagen type I, downregulate the expression of MMP-2, and thus exert its protective effect.