绞股蓝总皂苷对光老化人皮肤细胞p38MAPK和MMP-1的影响

Effect of Gypenosides on Expressions of p38MAPK and MMP-1 in Photoaging Skin Cells

目的：研究绞股蓝总皂苷对光老化人皮肤p38促分裂原活化蛋白激酶（p38MAPK）和基质金属蛋白酶-1（MMP-1）的影响。方法：制备空白大鼠血清和绞股蓝含药血清,将人角质形成细胞（HaCaT）分为4组：空白对照组（A）、UVB模型组（B）、空白血清组（C）和含药血清组（D）;将成纤维细胞（HSF）分为6组：空白对照组（Ⅰ）、UVA模型组（Ⅱ）、HaCaT培养上清液A组（Ⅲ）、HaCaT培养上清液B组（Ⅳ）、HaCaT培养上清液C组（Ⅴ）、HaCaT培养上清液D组（Ⅵ）。对HaCaT细胞B、C、D组和HSF细胞Ⅱ、Ⅲ、Ⅳ、Ⅴ、Ⅵ组分别用UVB和UVA照射,制备光老化细胞模型。将4组HaCaT细胞培养上清液分别加入HSF细胞的Ⅲ~Ⅵ组。采用RT-PCR和Western blotting方法检测Ⅰ~Ⅵ组细胞p38MAPK和MMP-1mRNA和蛋白的表达水平。结果：与Ⅰ组比较,Ⅱ组细胞p38MAPK和MMP-1 mRNA和蛋白的表达水平显著上调（P〈0.01）;与Ⅱ组比较,Ⅳ组p38MAPK和MMP-1mRNA和蛋白表达均上调（P〈0.01）,而Ⅲ组变化不显著;与Ⅳ组比较,Ⅵ组p38MAPK和MMP-1 mRNA和蛋白表达均显著下调（P〈0.01）,而Ⅴ组变化不显著。结论：光老化HaCaT细胞分泌细胞因子促进光老化HSF细胞p38MAPK和MMP-1 mRNA和蛋白的表达。绞股蓝总皂苷对光老化HaCaT细胞培养上清液作用的光老化HSF细胞中MAPK信号转导通路中相关基因和蛋白的表达具有抑制作用。

Objective： To study the effect of gypenosides-containing serum on the expressions of p38 mitogen-activated protein kinase （p38MAPK）and matrix metallo proteinase-1 （MMP-I）protein and mRNA in photoaging cells. Methods ： We made blank serum and gypenosides-containing serum and divided HaCaT into 4 groups： blank control group （ A ）, UVB model group （ B ）, blank serum group （ C ）, gypenosides-containing serum group （ D ） and divided HSF into 6 groups ： blank control group （ Ⅰ ）, UVA model group （ Ⅱ）, HaCaT supernatant A group （Ⅲ）, HaCaT supernatant B group （ Ⅳ ）, HaCaT supernatant C group （Ⅴ ）, HaCaT supernatant D group （ Ⅵ ）. We photoaged HaCaT B, C, D groups and HSF Ⅱ,Ⅲ,Ⅳ,Ⅴ,Ⅵ groups by UVB and UVA radiation. 4 groups HaCaT supernatant was put into HSF Ⅲ-Ⅵ groups in turn. The expression levels of p38MAPK and MMP-lmRNA and protein in each group were measured by RT-PCR and Western blotting method. Results： Compared with group Ⅰ , p38MAPK and MMP-lmRNA and protein expression of group Ⅱ were significantly higher （ P〈0.01 ）; compared with group Ⅱ , p38MAPK and MMP-lmRNA and protein expression in cells of group Ⅳ were significantly higher （P〈0.01）, the change of group Ⅲ were not significant; compared with group Ⅳ, p38MAPK and MMP- lmRNA and protein expression in cells of group Ⅵ were significantly lower （ P〈0.01 ）. The change of group V was not significant. Conclusion ： The photoaging HaCaT cell secrete cytokine to increase p38MAPK and MMP-lmRNA and protein expression in cell HSF. Gypenosides can inhibit p38MAPK and MMP-ImRNA and protein expression of photoaging HSF cells which were affected by photoaging HaCaT supernatant.