重组弹性蛋白的表征及在紫外诱导皮肤光老化过程中的作用  

Characterization of recombinant elastin and its roles in ultraviolet-induced skin photoaging

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作  者:张娅 周浩 王蓉 侯增淼 黄文涛 杨鹭 Ya Zhang;Hao Zhou;Rong Wang;Zengmiao Hou;Wentao Huang;Lu Yang(Xi’an De Nuohisi Medical Technology Co.,Ltd.,Xi’an,Shaanxi 710000,China)

机构地区:[1]西安德诺海思医疗科技有限公司,陕西西安710000

出  处:《日用化学工业(中英文)》2024年第9期1106-1116,共11页China Surfactant Detergent & Cosmetics

摘  要:采用基因工程技术构建重组弹性蛋白工程菌,再利用发酵纯化技术得到重组弹性蛋白,并经氨基酸N端测序、分子量测定、氨基酸组成、蛋白纯度分析、内毒素含量测定及其红外光谱和圆二色谱分析对其进行表征鉴定。通过测定重组弹性蛋白对L929小鼠胚胎成纤维细胞粘附能力和细胞活力影响,以及在中波紫外线(UVB)诱导L929细胞光老化模型中,考察重组弹性蛋白对Ⅰ型胶原蛋白(CollagenⅠ)、Ⅲ型胶原蛋白(CollagenⅢ)、弹性蛋白(Elastin)以及基质金属蛋白酶1(MMP-1)mRNA转录水平的影响,探究重组弹性蛋白在UVB诱导皮肤光老化过程中的作用。结果表明,获得的重组弹性蛋白与天然弹性蛋白二级结构相似;在0.1 mg/mL质量浓度下无细胞毒性且具有一定的细胞活力,显著促进L929细胞的粘附和增殖。此外,重组弹性蛋白可通过下调UVB辐射后L929细胞内MMP-1转录水平,提高细胞中的CollagenⅠ,CollagenⅢ和Elastin表达,对UVB诱导的L929细胞光损伤具有良好的保护作用,显著提升L929细胞的存活率。The recombinantelastin engineering bacteria was constructed by genetic engineering technology,and then recombinant elastin was obtained by fermentation purification technology.The recombinant elastin was characterized and identified by N-terminal sequencing of amino acids,molecular weight determination,analysis of amino acid composition,protein purity analysis,endotoxicity determination,infrared spectroscopy and circular dichroism analysis.To explore the roles of recombinant elastin in UVB-induced skin photoaging,the effects of recombinant elastin on the adhesion ability and cell viability of L929 mouse embryonic fibroblasts,as well as the effects of recombinant elastin on the expression levels of CollagenⅠ,CollagenⅢ,Elastin and matrix metalloproteinase 1(MMP-1),were investigated in the model of UVB-induced L929 cell photoaging.The results showed that the secondary structure of recombinant elastin was similar to that of natural elastin.The recombinant elastin showed certain cell viability and no cytotoxicity at 0.1 mg/mL,which significantly promoted the adhesion and proliferation of L929 cells.Additionally,recombinant elastin could also improve the mRNA transcription levels of CollagenⅠ,CollagenⅢand Elastin in UVB irradiated cells by down-regulating the expression of MMP-1 in UVB irradiated cells,thus protecting L929 cells from light damage.That was why recombinant elastin could improve the survival rate of photodamaged fibroblasts.

关 键 词:重组弹性蛋白 小鼠胚胎成纤维细胞 细胞外基质蛋白 光老化 紫外线(UVB) 

分 类 号:TQ658[化学工程—精细化工]

 

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