白桦脂醇对人皮肤成纤维细胞胶原蛋白及相关蛋白酶基因的调控机制研究

Regulative effect of collagen and related protease gene of betulin on human ESF-1 cells

目的研究自桦脂醇对人皮肤成纤维细胞（ESF-1）相关抗衰老基因mRNA的调控作用，探讨白桦脂醇对皮肤皱纹的修复机制。方法将细胞分为空白对照组，雌二醇组，白桦脂醇高、中、低剂量组。采用MTT法测定各组细胞活力，RT-PCR法测定各组细胞中I型及Ⅲ型胶原蛋白（collagen，Col）、基质金属蛋白酶抑制剂（tissueinhibitors of matrix metalloproteinase，TIMP）1和2、基质金属蛋白酶1（matrix metallo Droteinase1，MMP-1）mRNA的表达水平。结果与空白对照组比较，雌二醇组与白桦脂醇中剂量组细胞增殖率[（0．455±0．037）、（0．445±0．040）比（0．385±0．601）]增高（P〈0．05）；ColImRNA[（0．960±0．012）、（O．929±0．015）比（0．842±0．014）]，ColIIImRNA[（0．892±0．009）、（0．824±0．022）比（O．768±0．025）]、TIMP．1mRNA[（0．938±0．026）、（0．878±0．035）比（0．796±0．022）]、TIMP-2mRNAl（0．557±0．025）、（0．506±0．036）比（0．436±0．063）]表达水平均升高（P〈0．01）。结论白桦脂醇通过促进细胞增殖，提高Col合成及相关蛋白酶的表达等途径发挥对皱纹的修复作用。

Objective To explore the mechanism and regulative function of betulin on anti-aging genes related of normol human dermal fibroblasts （ESF-1）, and to reveal the repairing mechanism of betulin on wrinkles. Methods The cultured cell in vitro were divided into the control group, the estradiol group and the betulin high, medium and low dose groups. Competence of ESF-1 cells was detected by MTT. Expression of mRNA of collagen type I and III （Col I and Col III）, tissue inhibitors of matrix metalloproteinase 1 and 2 （TIMP-1 and TIMP-2）, matrix metalloproteinase 1 （MMP-1） was detected by RT-PCR. Results Compared with the control group, the proliferation of ESF-1 cells （0.455 ±0.037, 0.445 ± 0.040 vs. 0.385 ± 0.601） in the estradiol group and the medial-dose betulin group were increased （P〈0.05）. Compared with the control group, the expression of mRNA of Col I （0.960± 0.012, 0.929 ± 0.015 vs. 0.842 ±0.014）, Col III （0.892 ± 0.009, 0.824 ±0.022 vs. 0.768 ± 0.025）, TIMP-1 （0.938 ± 0.026, 0.878 ± 0.035 vs. 0.796 ± 0.022）, TIMP-2 （0.557 ± 0.025, 0.506 ± 0.036 vs. 0.436 ± 0.063） in the estradiol group and the medial-dose betulin group were increased （all P〈0.01）. Conclusion Betulin based on increasing the competence of ESF-1 cells, promoting collagen synthesis and the expression level of related-proteinase is to suspend the development of wrinkles.