摘要
目的研究补骨脂酚对中波紫外线(UVB)照射诱导的光老化人皮肤成纤维细胞(HSF)的保护作用。方法体外培养HSF细胞,分为空白组(正常培养,不进行UVB辐射)、UVB模型组(UVB辐射)、雌二醇组及补骨脂酚低(1μmol/L)、中(10μmol/L)、高(100μmol/L)剂量组。采用MTT法检测细胞活力;采用Western blot法检测细胞内Ⅰ型胶原蛋白(COLⅠ)、基质金属蛋白酶-1(MMP-1)和基质金属蛋白酶-3(MMP-3)的表达量;采用逆转录PCR法(RT-PCR)检测细胞COLⅠ、MMP-1 mRNA的表达水平。结果确定光老化HSF细胞模型的最佳UVB照射剂量为25 mJ/cm2。与空白组比较,补骨脂酚低、中、高剂量组细胞活力及MMP-1、MMP-3、COLⅠ分泌差异无统计学意义(P> 0.05);UVB模型组的细胞活力、COLⅠ的表达水平显著下降(P <0.05),MMP-1和MMP-3的表达水平显著上升(P <0.05);MMP-1 mRNA表达水平升高、COLⅠmRNA表达水平下降(P <0.05)。与UVB模型组比较,补骨脂酚低、中、高剂量组细胞活力、COLⅠ的表达量显著上升(P <0.05),MMP-1和MMP-3的表达水平显著下降(P <0.05),MMP-1 mRNA表达下降、COLⅠmRNA表达上升(P <0.05)。结论补骨脂酚能够促进HSF细胞COLⅠ表达,抑制MMP-1与MMP-3的表达,防护UVB对HSF细胞的损伤,发挥抗皮肤光老化的作用。
Objective To study the protective effect of bakuchiol on photoaging human skin fibroblasts(HSF) induced by ultraviolet radiation B(UVB) irradiation. Methods HSF cells were cultured in vitro and divided into blank group(normal culture, no UVB radiation), UVB model group(UVB radiation), estradiol group and bakuchiol low(1 μmol/L),medium(10 μmol/L) and high dose(100 μmol/L) groups. Cell viability was detected by MTT assay. The protein expressions of type Ⅰ collagen(COL Ⅰ), matrix metalloproteinase-1(MMP-1) and matrix metalloproteinase-3(MMP-3)were detected by Western blot. The expression levels of COL Ⅰ and MMP-1 m RNA were detected by reverse transcription polymerase chain reaction(RT-PCR). Results The optimal UVB radiation dose for photoaging HSF cell model was 25 mJ/cm2. Compared with the blank group, there was no significant difference in cell viability and secretion of MMP-1, MMP-3 and COL Ⅰ between the low, medium and high dose groups of bakuchiol(P > 0.05); the cell viability and COL Ⅰ expression level in the UVB model group decreased significantly(P < 0.05), the protein expression levels of MMP-1 and MMP-3 increased significantly(P < 0.05); the m RNA expression levels of MMP-1 and COL Ⅰ decreased significantly(P < 0.05). Compared with the UVB model group, the cell viability and COL Ⅰ expression in the low, middle and high dose groups of bakuchiol increased significantly(P < 0.05), the protein expression levels of MMP-1 and MMP-3 decreased significantly(P <0.05), the mRNA expression levels of MMP-1 and COL Ⅰ increased significantly(P < 0.05). Conclusion Bakuchiol can promote the expression of COL Ⅰ in HSFs, inhibit the expression of MMP-1 and MMP-3,protect UVB from damage to HSFs and exert anti-photoaging effects on skin.
引文
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