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

doi:10.3969/j.issn.2097-2806.2024.09.012

Abstract

Abstract:

The recombinant-elastin 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 Collagen Ⅰ, Collagen Ⅲ, 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 Collagen Ⅰ, Collagen Ⅲ and 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.

Key words: recombinant elastin, mouse embryonic fibroblast, extracellular matrix protein, photoaging, ultraviolet light

CLC Number:

TQ658

Ya Zhang, Hao Zhou, Rong Wang, Zengmiao Hou, Wentao Huang, Lu Yang. Characterization of recombinant elastin and its roles in ultraviolet-induced skin photoaging[J].China Surfactant Detergent & Cosmetics, 2024, 54(9): 1106-1116.

Figures/Tables 12

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References 25

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氨基酸名称	理论氨基酸个数	样品考察
氨基酸名称	理论氨基酸个数	实测摩尔数	实测氨基酸个数	误差百分比/%	实测摩尔百分比/%
Asp（天冬氨酸）	0	—	—	—	—
Glu（谷氨酸）	0	—	—	—	—
Ser（色氨酸）	0	—	—	—	—
Gly（甘氨酸）	95	4 325.46	92.45	-2.67	32.78
His（组氨酸）	0	—	—	—	—
Arg（精氨酸）	0	—	—	—	—
Thr（苏氨酸）	0	—	—	—	—
Ala（丙氨酸）	46	1 872.70	40.03	-12.99	14.19
Pro（脯氨酸）	49	2 277.15	48.67	-0.67	17.26
Tyr（酪氨酸）	0	—	—	—	—
Val（缬氨酸）	92	4 718.73	100.85	9.62	35.76
Met（蛋氨酸）	0	—	—	—	—
Cys-Cys（半胱氨酸）	0	—	—	—	—
Lle（异亮氨酸）	0	—	—	—	—
Leu（亮氨酸）	0	—	—	—	—
Phe（苯丙氨酸）	0	—	—	—	—
Trp（色氨酸）	0	—	—	—	—
Lys（赖氨酸）	0	—	—	—	—

泳道	纯度/%
泳道2	97.9
泳道3	97.5
泳道4	97.0
泳道5	97.5
泳道6	98.0
泳道7	98.5
平均值	97.7

样品名称	重组弹性蛋白	2λ/重组弹性蛋白	2λ/ 检查用水	检查用水
1	-/-	+/+	+/+	-/-
2	-/-	+/+
3	-/-	+/+

组别	照射时间/min	A值	存活率/%
对照	0	0.66±0.031	100
UVB造模	1	0.65±0.019	98
	3	0.59±0.011	89
	5	0.48±0.017 ^*	73
	10	0.30±0.012 ^**	45
	15	0.24±0.014 ^**	36
	30	0.21±0.005 ^**	32

组别	A值	存活率/%
对照	0.31±0.028	100
UVB模型组	0.23±0.008 ^**	74
UVB+0.01 mg/mL重组弹性蛋白	0.25±0.016 ^*	81
UVB+0.05 mg/mL重组弹性蛋白	0.26±0.008 ^*	84
UVB+0.1 mg/mL重组弹性蛋白	0.29±0.011	94
UVB+0.25 mg/mL重组弹性蛋白	0.25±0.019 ^*	81

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

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