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Objective To investigate the possible mechanisms using different chemicals to improve the skin aging by the effect of estrogen,vitamin E,vitamin C on the transcriptional level of MMP-1 and HAS-2 mRNAs in cultured human skin fibroblast in vitro.Methods The different concentrations of estrogen,vitamin E,vitamin C were put into cultured human skin fibroblasts.After 24 hours,RT-PCR was used to detect the effects of the different chemicals on the transcriptional levels of the HAS-2 and MMP-1 mRNAs in fibroblasts.Results The effects of estrogen and vitamin C on the MMP-1 mRNA had significant difference between the high dose group and blank control group,and between the low dose group and blank control group (P<0.05) ; the difference between the low dose group and high dose group was not statistically significant (P>0.05).The effects of vitamin E and vitamin C on Has-2 mRNA also had significant difference between the high dose group and the blank control group,and between the low dose group and blank control group (P<0.05),but the difference between the low dose group and high dose group was not statistically significant (P>0.05).Conclusions Estrogen and vitamin C can reduce the transcription of MMP-1 gene,which decreases the degradation of collagen in the skin; vitamin E and vitamin C can increase the transcription of HAS-2 gene,which may possibly increase the synthesis of hyaluronic acid to alleviate the skin aging.
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Objective To investigate the influences of lysozyme on the mRNA expressions of MMP-1,-12 and lysyl oxidase(LOX)in cultured fibroblasts in vitro.Methods Primarily cultured fibroblasts isolated from human skin were treated with three concentrations(0.1×10~(-8),1×10~(-7)mol/L)of lysozyme followed by another 24-hour cuhure.Subsequently,total RNA was extracted from the fibroblasts and subjected to RT-PCR for the detection of MMP-1,-12 and LOX mRNA.Results There was a significant difference in the mRNA expressions of MMP-1,-12 and LOX among the fibroblasts treated with the three concentrations of lysozyme (F=6.98,4.44,5.24,respectively,all P<0.05).SNK-q test showed that untreated fibroblasts differed signifi-cantly from those treated with lysozyme of 1×10~(-7) mol/L in the mRNA expression of MMP-1 and MMP-12 (P<0.05),and from those treated with iysozyme of 1×10~(-7) mol/L.and 1×10~(-8)mol/L in the mRNA expres-sion of LOX(both P<0.05),whereas no significant difference was ohserved between fibroblasts treated with lysozyme of 1×10~(-8) mol/L and untreated fibrohlasts or those with lysozyme of 1 x 10~(-7)mol/L in the mRNA expression of MMP-1 and MMP-12.or between fibroblasts treated with lysozyme of 1 x 10~(-8)mol/L and those with that of 1×10~(-7) mol/L in the expression of LOX (all P>0.05).Conclusions Lysozyme upregulates the mRNA expression of MMP-1 and MMP-12 but downregulates the mRNA expression of LOX in cultured fibro-blasts in vitro.
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Objective To explore the mechanism of vitamin E on delaying skin aging by observ-ing the expression of hyaluronic acid synthetase-2 (HAS-2) in human dermal fibroblasts in vitro. Methods Human skin fibroblasts were cultured in vitro, and these fibroblast cells were then divided into 3 groups: different concentration of vitamin E (0, 0.1 × 10-10, 1 ×109mol/L) was added in the medium in the different group. 24 hours later, the fibroblasts were collected, RNAs extracted, and then amplified by RT-PCR. The PCR product was determined by agarose gel electrophoresis, to analyze the level of HAS-2 mRNA expression. Results RT-PCR showed the lever of HAS-2 mRNA was higher in the low-dose group than the control group, with significant difference (P<0.05) ; the lever of HAS-2 mRNA was higher in the high-dose group than control group, with significant difference (P<0.05). There was no significant difference in the lever of HAS-2 mRNA between the low-dose group and the high-dose group. Conclusions Vitamin E can enhance the expression of hyaluronic acid synthetase-2 mRNA, may increase the synthesis of HAS in skin fibroblasts and increase water content in the skin, so that it might reverse or delay the skin aging.