Comparative effects of biodynes, tocotrienol-rich fraction, and tocopherol in enhancing collagen synthesis and inhibiting collagen degradation in stress-induced premature senescence model of human diploid fibroblasts

Abstract

Biodynes, tocotrienol-rich fraction (TRF), and tocopherol have shown antiaging properties. However, the combined effects of these compounds on skin aging are yet to be investigated. This study aimed to elucidate the skin aging effects of biodynes, TRF, and tocopherol on stress-induced premature senescence (SIPS) model of human diploid fibroblasts (HDFs) by determining the expression of collagen and MMPs at gene and protein levels. Primary HDFs were treated with biodynes, TRF, and tocopherol prior to hydrogen peroxide (H2O2) exposure. The expression of COL1A1, COL3A1, MMP1, MMP2, MMP3, and MMP9 genes was determined by qRT-PCR. Type I and type III procollagen proteins were measured by Western blotting while the activities of MMPs were quantified by fluorometric Sensolyte MMP Kit. Our results showed that biodynes, TRF, and tocopherol upregulated collagen genes and downregulated MMP genes (P < 0.05). Type I procollagen and type III procollagen protein levels were significantly increased in response to biodynes, TRF, and tocopherol treatment (P < 0.05) with reduction in MMP-1, MMP-2, MMP-3, and MMP-9 activities (P < 0.05). These findings indicated that biodynes, TRF, and tocopherol effectively enhanced collagen synthesis and inhibited collagen degradation and therefore may protect the skin from aging.

Figure 1

Effects of biodynes, TRF, and tocopherol on COL1A1 gene and procollagen type I protein expression in HDFs. COL1A1 expression (a). Biodynes, TRF, tocopherol, and BTT significantly increased COL1A1 expression compared to SIPS. Procollagen type I protein expression (b). TRF, tocopherol, and BTT significantly increased the expression of procollagen type I. ^aDenotes P < 0.05 compared to SIPS, ^bdenotes P < 0.05 compared to biodynes, ^cdenotes P < 0.05 compared to TRF, and ^ddenotes P < 0.05 compared to tocopherol. Data are presented as the mean of three experiments ±S.D, n = 6.

Figure 2

Effects of biodynes, TRF, and tocopherol on COL3A1 gene and procollagen type III protein expression in HDFs. COL3A1 expression (a). Biodynes and TRF significantly increased COL3A1 expression compared to SIPS. Procollagen type III protein expression (b). Biodynes, TRF, tocopherol, and BTT significantly increased the expression of procollagen type III. ^aDenotes P < 0.05 compared to SIPS, ^bdenotes P < 0.05 compared to biodynes. Data are presented as the mean of three experiments ±S.D, n = 6.

Figure 3

Effects of biodynes, TRF, and tocopherol on MMP1 gene expression and MMP-1 activity in HDFs. MMP1 expression (a). TRF, tocopherol, and BTT significantly downregulated MMP1 compared to SIPS. MMP-1 activity (b). BTT decreased the MMP-1 activity compared to SIPS. ^aDenotes P < 0.05 compared to SIPS. Data are presented as the mean of three experiments ±S.D, n = 6.

Figure 4

Effects of biodynes, TRF, and tocopherol on MMP2 gene expression and MMP-2 activity in HDFs. MMP2 expression (a). TRF, tocopherol and BTT downregulated MMP2 compared to SIPS. MMP-2 activity (b). Biodynes, TRF, tocopherol, and BTT significantly decreased, the MMP-2 activity compared to SIPS. ^aDenotes P < 0.05 compared to SIPS. Data are presented as the mean of three experiments ±S.D, n = 6.

Figure 5

Effects of biodynes, TRF, and tocopherol on MMP3 gene expression and MMP-3 activity in HDFs. MMP3 expression (a). Biodynes, TRF, tocopherol, and BTT significantly downregulated MMP3 compared to SIPS. MMP-3 activity (b). Biodynes, TRF, tocopherol, and BTT significantly decreased MMP-3 activity. ^aDenotes P < 0.05 compared to SIPS. Data are presented as the mean of three experiments ±S.D, n = 6.

Figure 6

Effects of biodynes, TRF and tocopherol on MMP9 gene expression and MMP-9 activity in HDFs. MMP9 expression (a). Biodynes, TRF, tocopherol and BTT significantly down regulated MMP9 expression compared to SIPS. MMP-9 activity (b). Biodynes, tocopherol and BTT significantly decreased MMP-9 activity. ^aDenotes P < 0.05 compared to SIPS, ^adenotes P < 0.05 compared to biodynes, ^cdenotes P < 0.05 compared to TRF, ^ddenotes P < 0.05 compared to tocopherol. Data are presented as the mean of three experiments ±S.D, n = 6.