Effects Of Different Drugs On The Expression Of Matrix Metalloproteinase-1,Matrix Metalloproteinase-12,Lysyl Oxidase Gene In Human Dermal Fibroblasts In Vitro

Objective:Many enzymes produced by the body play important roles during the skin aging, which are one of the important reasons and their pathophy-siological mechanisms are closely related to skin aging. The purpose of this study is to expore the mechanism of lysozyme (LZM), vitamin E (Vit-E), estrogen (ESG) and basic fibroblast growth factor (bFGF) on delaying skin aging by observing the expression of matrix metalloproteinase-1,12(MMP-1,12) and lysine oxidase(LOX) mRNA in human skin fibroblasts (HSF) in vitro.Methods:Human skin fibroblast were cultured in vitro using the method of enzyme digestion,these fibroblast cells were then divided into 3 groups, lower dimensional LZM, Vit-E, ESG and bFGF were added in the medium Separately in the second group, higher dimensional LZM, Vit-E, ESG and bFGF were added in the medium Separately in the third group. A few days later, we collected the fibroblast, which was index number growth phase form different groups, extracted RNA, then amplified by RT-PCR. The PCR product was determined by agorose electrophoresis, to analysis the level of MMP-1,MMP-12,LOX mRNA expression.Results:1. LZM group (0 mol/L, 1×10-8 mol/L, 1×10-7mol/L):RT-PCR showed that no significant difference(P＞0.05) was found in MMP-1,MMP-12 mRNA expression as between the low-dose group and control group as well as between the low-dose group and the high-dose group; MMP-1,MMP-12 mRNA expression was significantly lower(P＜0.05)in the high-dose group than the control group; LOX mRNA expression was higher(P＜0.05) in low-dose group than control group and the high-dose group than the control group; No significant difference(P＞0.05) was found in LOX mRNA expression as between the low-dose group and the high-dose group.2. Vit-E group (0 mol/L, 1×10-5 mol/L, 1×10-4 mol/L):RT-PCR showed that the difference of the expression MMP-12 and LOX mRNA among control group, low-dose group and high-dose group was statistically significant (F=7.410/5.820, P=0.008/0.017), SNK-q test showed that no significant difference(P＞0.05) was found in MMP-12 and LOX mRNA expression as between the low-dose group and control group as well as between the low-dose group and the high-dose group; MMP-12 mRNA expression was significantly lower(P<0.05)in the high-dose group than the control group. LOX mRNA expression was significantly higher(P<0.05)in the high-dose group than the control group.3. ESG group(0 mol/L, 1×10-5 mol/L, 1×10-4 mol/L):RT-PCR showed that there was no significant difference of the expression MMP-12 mRNA among control group, low-dose group and high-dose group(F=3.711, P=0.056). The difference of the expression LOX mRNA among these three groups was statistically significant (F=4.337,P=0.038). SNK-q test showed that no significant difference (P>0.05) was found in LOX mRNA expression between the low-dose group and control group as well as between the low-dose group and the high-dose group; LOX mRNA expression was significantly higher(P＜0.05)in the high-dose group than the control group.4. bFGF group(0μg/L,10μg/L,50μg/L):RT-PCR showed that the difference of the expression MMP-1 among control group, low-dose group and high-dose group was statistically significant (F=4.809, P=0.029), and no significant difference of the expression MMP-12 and LOX mRNA among control group, low-dose group and high-dose group(F=3.429/2.173, P=0.066/0.157). SNK-q test showed that compared with the control group, the difference of High-dose bFGF on level of MMP-1 mRNA expression was statistically significant(P<0.05). the difference of low-dose bFGF on level of MMP-1 mRNA expression was no statistically significant (P>0.05).Conclusion:1. lysozyme can decrease the level of MMP-1,MMP-12 mRNA expression, and enhance the level of LOX mRNA expression.2. Vit E can decrease the level of MMP-12 mRNA expression,and enhance the level of LOX mRNA expression.3. Estrogen can enhance the level of LOX mRNA expression.4. bFGF can decrease the level of MMP-1 mRNA expression.