驴骨胶原蛋白成分分析及促进皮肤修复活性的研究

樊雨梅 史传超 帖航 张筱梒 赵海晴 廖峰

樊雨梅,史传超,帖航,等. 驴骨胶原蛋白成分分析及促进皮肤修复活性的研究[J]. 食品工业科技,2021,42(23):341−349. doi:  10.13386/j.issn1002-0306.2021010211
引用本文: 樊雨梅,史传超,帖航,等. 驴骨胶原蛋白成分分析及促进皮肤修复活性的研究[J]. 食品工业科技,2021,42(23):341−349. doi:  10.13386/j.issn1002-0306.2021010211
FAN Yumei, SHI Chuanchao, TIE Hang, et al. Study on the Nutritional Ingredients and the Effect on the Repair of Damaged Skin Barrier Function of Donkey Bone Collagen[J]. Science and Technology of Food Industry, 2021, 42(23): 341−349. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2021010211
Citation: FAN Yumei, SHI Chuanchao, TIE Hang, et al. Study on the Nutritional Ingredients and the Effect on the Repair of Damaged Skin Barrier Function of Donkey Bone Collagen[J]. Science and Technology of Food Industry, 2021, 42(23): 341−349. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2021010211

驴骨胶原蛋白成分分析及促进皮肤修复活性的研究

doi: 10.13386/j.issn1002-0306.2021010211
基金项目: 山东省泰山产业领军人才传统产业创新类项目(TSCY20170233)
详细信息
    作者简介:

    樊雨梅(1992−),女,硕士,工程师,研究方向:天然产物功效成分,E-mail:bzguo9@163.com

    通讯作者:

    廖峰(1981−),男,博士,正高级工程师,研究方向:天然产物功效研究与应用,E-mail:liaofengcd@163.com

  • 中图分类号: TS251.9

Study on the Nutritional Ingredients and the Effect on the Repair of Damaged Skin Barrier Function of Donkey Bone Collagen

  • 摘要: 为从分子水平探究驴骨胶原蛋白的功效物质基础和驴骨胶原蛋白对受损皮肤屏障的修复作用,本实验采用超高效液相色谱综合高分辨率轨道分析质谱(UPLC-Orbitrap MS)技术定性分析驴骨胶原蛋白的有效成分,用5-溴脱氧尿嘧啶核苷(5-bromodeoxyuridine, BrdU)渗入法和细胞划痕实验检测驴骨胶原蛋白对角质形成细胞增殖和迁移的影响,以探究其对皮肤的修复作用。结果表明:驴骨胶原蛋白的蛋白含量为57.5%,具有典型的胶原氨基酸组成。驴骨胶原蛋白含有较为丰富的氨基酸、维生素、糖类和脂质等活性成分。0.15 g/mL的驴骨胶原蛋白促进人表皮角质形成细胞(HaCaT细胞)增殖和迁移的效果与5%胎牛血清类似,显著优于空白对照组(P<0.05)。最终推断驴骨胶原蛋白能够加速表皮屏障修复,其机制可能与促进角质形成细胞增殖和迁移有关。
  • 图  1  驴骨胶原蛋白总离子流色谱图

    Figure  1.  The total ion chromatogram of donkey bone collagen

    注:a:正离子模式;b:负离子模式。

    图  2  驴骨胶原蛋白对HaCaT细胞存活率的影响

    Figure  2.  Effects of donkey bone collagen on the survival rate of HaCaT cells

    注:*表示与空白对照组相比差异显著,P<0.05;**表示与空白对照组相比差异极显著,P<0.01;图3~图4同。

    图  3  驴骨胶原蛋白(a)和各处理组(b)对HaCaT细胞增殖的影响

    Figure  3.  Effects of donkey bone collagen(a)and different treatments(b)on the proliferation of HaCaT cells

    图  4  驴骨胶原蛋白和各处理组对HaCaT细胞迁移的影响

    Figure  4.  Effects of donkey bone collagen and different treatments on the migration of HaCaT cells

    注:图A为划痕实验图像;图B为对细胞迁移率的影响结果。

    表  1  流动相洗脱程序

    Table  1.   Elution procedure of mobile phase

    时间(min)流速(mL/min)A相(%)B相(%)
    00.3955
    10.3955
    20.36040
    70.32080
    110.3595
    15.010.3595
    15.50.3955
    19.50.3955
    下载: 导出CSV

    表  2  驴骨胶原蛋白的氨基酸组成及含量

    Table  2.   The amino acid composition and content of donkey bone collagen

    氨基酸种类含量(%)氨基酸种类含量(%)
    天门冬氨酸6.708半胱氨酸0.000
    谷氨酸12.616缬氨酸4.064
    丝氨酸3.297蛋氨酸0.139
    组氨酸1.097苯丙氨酸2.941
    甘氨酸24.037异亮氨酸1.690
    苏氨酸2.360亮氨酸4.155
    精氨酸8.780赖氨酸4.580
    丙氨酸9.793脯氨酸12.717
    酪氨酸0.154
    下载: 导出CSV

    表  3  驴骨胶原蛋白LC-MS全谱鉴定

    Table  3.   Compound identification of donkey bone collagen by LC-MS

    序号名称理论分子量分类分子式保留时间(min)
    1γ-氨基丁酸103.0633氨基酸及其衍生物类C4H9NO20.902
    2L-(-)-丝氨酸105.0426氨基酸及其衍生物类C3H7NO30.876
    3L-(-)-苏氨酸119.0582氨基酸及其衍生物类C4H9NO30.882
    4牛磺酸125.0147氨基酸及其衍生物类C2H7NO3S0.895
    5L-焦谷氨酸129.0426氨基酸及其衍生物类C5H7NO31.425
    6L-(+)-亮氨酸131.0946氨基酸及其衍生物类C6H13NO21.738
    7L-(+)-天冬氨酸133.0375氨基酸及其衍生物类C4H7NO40.893
    8DL-谷氨酰胺146.0691氨基酸及其衍生物类C5H10N2O30.883
    9DL-赖氨酸146.1055氨基酸及其衍生物类C6H14N2O20.816
    10DL-谷氨酸147.0532氨基酸及其衍生物类C5H9NO40.902
    11DL-组氨酸155.0695氨基酸及其衍生物类C6H9N3O20.839
    12DL-苯丙氨酸165.0790氨基酸及其衍生物类C9H11NO22.956
    13脱氨基酪氨酸166.0630氨基酸及其衍生物类C9H10O34.346
    14DL-精氨酸174.1117氨基酸及其衍生物类C6H14N4O20.841
    15DL-酪氨酸181.0739氨基酸及其衍生物类C9H11NO31.534
    16DL-色氨酸204.0899氨基酸及其衍生物类C11H12N2O23.712
    172-氯苯丙氨酸199.0400内标C9H10ClNO23.725
    18黄嘌呤152.0334嘌呤类C5H4N4O21.409
    19D-木糖酸166.0480糖类C5H10O60.922
    20己糖180.0634糖类C6H12O60.975
    21己糖醇182.0790糖类C6H14O60.882
    22D-葡萄糖酸196.0583糖类C6H12O70.914
    23N-乙酰-α-氨基葡萄糖-1-磷酸酯301.0563糖类C8H16NO9P0.917
    24海藻糖342.1162糖类C12H22O110.912
    25维生素B5219.1110维生素类C9H17NO53.511
    26DL-乳酸90.0317有机酸类C3H6O31.112
    27苹果酸134.0215有机酸类C4H6O50.983
    28水杨酸138.0317有机酸类C7H6O33.967
    29环己基乙酸142.0990有机酸类C8H14O25.275
    30丁酰乙酸乙酯158.0940有机酸类C8H14O35.342
    31焦碳酸二乙酯162.0530有机酸类C6H10O50.884
    32尿酸168.0283有机酸类C5H4N4O30.986
    33乙酰丙酸丁酯172.1100有机酸类C9H16O35.474
    34马尿酸179.0582有机酸类C9H9NO34.026
    35柠檬酸192.0270有机酸类C6H8O70.998
    36银杏酸346.2510有机酸类C22H34O39.063
    37月桂酸200.1776脂质类C12H24O29.457
    38癸酸172.1460脂质类C10H20O28.054
    39杜鹃花酸188.1050脂质类C9H16O44.803
    405-羟基癸酸188.1410脂质类C10H20O36.078
    41癸二酸202.1210脂质类C10H18O47.416
    42乙酰肉碱203.1158脂质类C9H17NO44.169
    4312-羟基月桂酸216.1730脂质类C12H24O37.406
    44甘油磷酸246.0505脂质类C6H15O8P0.910
    45肉豆蔻酸乙酯256.2402脂质类C16H32O28.339
    46棕榈酸256.2402脂质类C16H32O212.737
    473-氧代棕榈酸270.2195脂质类C16H30O39.208
    48亚油酸280.2402脂质类C18H32O211.814
    49油酸282.2559脂质类C18H34O28.654
    50硬脂酸284.2715脂质类C18H36O29.711
    51十六烷二酸286.2144脂质类C16H30O46.508
    52肉豆蔻酯硫酸盐294.1865脂质类C14H30O4S15.020
    5313-KODE294.2195脂质类C18H30O38.965
    54蓖麻油酸298.2508脂质类C18H34O311.768
    5512-羟基硬脂酸300.2664脂质类C18H36O39.809
    56花生四烯酸304.2402脂质类C20H32O29.112
    57二高-γ-亚麻酸306.2559脂质类C20H34O212.294
    58(+/-)12(13)-DiHOME314.2457脂质类C18H34O47.382
    599,10-羟基硬脂酸316.2614脂质类C18H36O47.987
    6015S-羟基二十碳三烯酸322.2508脂质类C20H34O39.160
    61(15Z)-9,12,13-三羟基-15-十八碳烯酸330.2406脂质类C18H34O55.345
    62(10E)-9,12,13-三羟基-10-十八碳烯酸330.2406脂质类C18H34O55.880
    63二十碳五烯酸乙酯330.2559脂质类C22H34O211.815
    6413,14-二氢-15-酮前列腺素A2334.2144脂质类C20H30O45.536
    65前列腺素A1336.2301脂质类C20H32O46.682
    66白三烯B4336.2301脂质类C20H32O46.743
    67N-油酰甘氨酸339.2773脂质类C20H37NO310.968
    68十二烷酰肉碱343.2723脂质类C19H37NO49.873
    69前列腺素E2352.225脂质类C20H32O55.535
    706β-前列腺素I1354.2406脂质类C20H34O55.807
    71前列腺素E1354.2406脂质类C20H34O55.594
    72前列腺素F1α356.2563脂质类C20H36O55.938
    7313,14-二氢前列素E1356.2563脂质类C20H36O55.689
    74甘膦酸-N-棕榈酰乙醇胺453.2855脂质类C21H44NO7P8.299
    751-十六烷酰基-sn-甘油-3-磷酸乙醇胺453.2855脂质类C21H44NO7P8.067
    762-亚油酰-sn-甘油-3-磷酸乙醇胺477.2855脂质类C23H44NO7P7.866
    771-油酰-sn-甘油-3-磷酸乙醇胺479.3012脂质类C23H46NO7P8.654
    781-硬脂酰-sn-甘油-3-磷酸乙醇胺481.3168脂质类C23H48NO7P9.420
    791-十六烷酰-sn-甘油-3-磷酸胆碱495.3325脂质类C24H50NO7P9.123
    801-花生四烯酰-sn-甘油-3-磷酸乙醇胺501.2855脂质类C25H44NO7P7.715
    81油酰溶血磷脂酰胆碱521.3481脂质类C26H52NO7P10.836
    821-(9z-十六烷酰基)-2-(9z,12z-十八烷酰基)-sn-甘油590.4910脂质类C37H66O513.840
    83丙二醛72.0211其他C3H4O20.884
    84磷酸97.9769其他H3O4P0.930
    85尿囊素158.0440其他C4H6N4O30.953
    86反式香叶酸168.1150其他C10H16O28.864
    87肌肽226.1070其他C9H14N4O30.794
    88雄甾烯醇274.2300其他C19H30O5.589
    下载: 导出CSV
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出版历程
  • 收稿日期:  2021-01-27
  • 网络出版日期:  2021-10-22
  • 刊出日期:  2021-12-01

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