Nuryana Ch Tri, Haryana Sofia Mubarika, Wirohadidjojo Yohanes Widodo, Arfian Nur
Doctoral program of Medicine and Health, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia.
Department of Histology and Cell Biology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia.
J Stem Cells Regen Med. 2020 May 27;16(1):26-31. doi: 10.46582/jsrm.1601005. eCollection 2020.
Ultraviolet radiation induces skin photoaging by increasing matrix metalloproteinase-1 (MMP-1). MMP-1 degrades type I and III collagen that comprise the dermal connective tissue. mucous (AFM) is a natural remedy that has protective effects on fibroblasts and collagen. To investigate the effects of AFM on cell viability and collagen deposition in UVB-irradiated human fibroblast culture. The mucous was extracted from 50 snails that were stimulated by a 5-10 Volt electricity shock for 30-60 seconds and converted into powder by the freeze-drying process. The human dermal fibroblast culture was divided into six groups: group 1 were normal fibroblasts without UVB irradiation as normal control, groups 2-5 consisted of 100 mJ/cm UVB-irradiated fibroblasts. Group 2 had no treatment as negative control, group 3 was treated by PRP 10% as positive control group and groups 4-6 were treated by various concentrations of AFM (3.9; 15.625 and 62.5 μg/mL). At the end of the experiment, the proliferation was assessed with MTT assay, furthermore collagen deposition was measured by Sirius red assay. Real Time-PCR (RT-PCR) was performed to quantify Coll I, Coll III and MMP-1 mRNA expression, then to measured COL 1/COL III ratio. UVB induced significant lower viability, upregulated MMP-1 and downregulated COL I and COL III mRNA expressions. Meanwhile AFM treated groups demonstrated higher cell viability with downregulation of MMP-1 and upregulation of COL I and COL III mRNA expressions. The ratio of COL I/ III expression was significantly (<0.05) lower in the AFM treated groups compared to the UVB group. Among AFM treated groups, administration of 62.5 μg/mL AFM represented the best result. AFM may ameliorate viability of UVB-irradiated human fibroblast culture which associates with downregulating MMP-1, upregulating COL I and Col III, and reducing COL I/III ratio.
紫外线辐射通过增加基质金属蛋白酶-1（MMP-1）诱导皮肤光老化。MMP-1降解构成真皮结缔组织的I型和III型胶原蛋白。 黏液（AFM）是一种对成纤维细胞和胶原蛋白具有保护作用的天然疗法。为了研究AFM对紫外线B（UVB）照射的人成纤维细胞培养物中细胞活力和胶原蛋白沉积的影响。从50只蜗牛中提取黏液，这些蜗牛经5-10伏电击刺激30-60秒，然后通过冷冻干燥过程转化为粉末。人真皮成纤维细胞培养物分为六组：第1组为未接受UVB照射的正常成纤维细胞作为正常对照，第2-5组为由100 mJ/cm² UVB照射的成纤维细胞组成。第2组未进行处理作为阴性对照，第3组用10%富血小板血浆（PRP）处理作为阳性对照组，第4-6组用不同浓度的AFM（3.9；15.625和62.5 μg/mL）处理。在实验结束时，用MTT法评估细胞增殖，此外用天狼星红法测量胶原蛋白沉积。进行实时聚合酶链反应（RT-PCR）以定量I型胶原（Coll I）、III型胶原（Coll III）和MMP-1信使核糖核酸（mRNA）表达，然后测量COL 1/COL III比率。UVB诱导显著较低的活力，上调MMP-1并下调COL I和COL III mRNA表达。同时，AFM处理组表现出较高的细胞活力，MMP-1下调，COL I和COL III mRNA表达上调。与UVB组相比，AFM处理组中COL I/III表达比率显著降低（<0.05）。在AFM处理组中，给予62.5 μg/mL AFM的效果最佳。AFM可能改善UVB照射的人成纤维细胞培养物的活力，这与下调MMP-1、上调COL I和Col III以及降低COL I/III比率有关。
