Zhang Jia-an, Yin Zhi, Ma Li-wen, Yin Zhi-qiang, Hu Yan-yan, Xu Yang, Wu Di, Permatasari Felicia, Luo Dan, Zhou Bing-rong
Department of Dermatology, the First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, P. R. China.
PLoS One. 2014 Jun 20;9(6):e99703. doi: 10.1371/journal.pone.0099703. eCollection 2014.
This study was aimed to evaluate the anti-photoaging effects of baicalin on Ultraviolet B (UVB)-induced photoaging in the dorsal skin of hairless mice and premature senescence in human dermal fibroblasts.
We established in vivo and in vitro photoaging models by repeated exposures to UVB irradiation. By HE staining, masson staining, immunohistostaing and real-time RT-PCR, we analyzed epidermal thickness, collagen expression and the mRNA and protein levels of type I collagen, type III collagen, interstitial collagenase (MMP-1 and MMP-3) in UVB exposed dorsal mice skin. The aging condition in human dermal fibroblasts was determined by senescence-associated β-galactosidase (SA-β-gal) staining. Cell viability was determined using the Cell Counting Kit-8 (CCK-8). The G1 phase cell growth arrest was analyzed by flow cytometry. The senescence-related protein levels of p16INK-4a, p21WAF-1, and p53 and protein levels of phosphorylated histone H2AX were estimated by Western blotting.
Topically application of baicalin treatment reduced UVB-induced epidermal thickening of mouse skin and also result in an increase in the production of collagen I and III, and a decrease in the expression of MMP-1 and MMP-3. Compared with the UVB-irradiated group, we found that the irradiated fibroblasts additionally treated with baicalin demonstrated a decrease in the expression of SA-β-gal, a increase in the cell viability, a decrease in the G1 phase cell proportion, a downregulation in the level of senescence-associated and γ-H2AX proteins. However, Baicalin had no difference in the normal fibroblasts without UVB irradiation and long-term Baicalin incubation of UVB-SIPS fibroblasts gave no effects on the cell proliferation.
Taken together, these results suggest that baicalin significantly antagonizes photoaging induced by UVB in vivo and in vitro, indicating the potential of baicalin application for anti-photoaging treatment.
本研究旨在评估黄芩苷对紫外线B(UVB)诱导的无毛小鼠背部皮肤光老化及人真皮成纤维细胞早衰的抗光老化作用。
通过反复暴露于UVB辐射建立体内和体外光老化模型。通过苏木精-伊红(HE)染色、马松染色、免疫组织化学和实时逆转录-聚合酶链反应(RT-PCR),分析UVB照射的小鼠背部皮肤的表皮厚度、胶原蛋白表达以及I型胶原蛋白、III型胶原蛋白、间质胶原酶(MMP-1和MMP-3)的mRNA和蛋白水平。通过衰老相关β-半乳糖苷酶(SA-β-gal)染色确定人真皮成纤维细胞的衰老状况。使用细胞计数试剂盒-8(CCK-8)测定细胞活力。通过流式细胞术分析G1期细胞生长停滞情况。通过蛋白质印迹法评估衰老相关蛋白p16INK-4a、p21WAF-1和p53的水平以及磷酸化组蛋白H2AX的蛋白水平。
局部应用黄芩苷治疗可减轻UVB诱导的小鼠皮肤表皮增厚,还可导致I型和III型胶原蛋白生成增加,以及MMP-1和MMP-3表达减少。与UVB照射组相比,我们发现用黄芩苷额外处理的照射后成纤维细胞SA-β-gal表达降低、细胞活力增加、G1期细胞比例降低、衰老相关蛋白和γ-H2AX蛋白水平下调。然而,黄芩苷对未接受UVB照射的正常成纤维细胞没有差异,且对UVB-慢照射后成纤维细胞长期进行黄芩苷孵育对细胞增殖没有影响。
综上所述,这些结果表明黄芩苷在体内和体外均能显著拮抗UVB诱导的光老化,表明黄芩苷在抗光老化治疗中的应用潜力。