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鲟鱼软骨制品的工艺优化及其对人软骨细胞炎症的调控

刘恒阁 徐新星 白帆 汪金林 高瑞昌 刘康 赵元晖

刘恒阁,徐新星,白帆,等. 鲟鱼软骨制品的工艺优化及其对人软骨细胞炎症的调控[J]. 食品工业科技,2022,43(15):165−174. doi:  10.13386/j.issn1002-0306.2021100051
引用本文: 刘恒阁,徐新星,白帆,等. 鲟鱼软骨制品的工艺优化及其对人软骨细胞炎症的调控[J]. 食品工业科技,2022,43(15):165−174. doi:  10.13386/j.issn1002-0306.2021100051
LIU Hengge, XU Xinxing, BAI Fan, et al. Optimization of Sturgeon Cartilage Products and Its Regulation on Human Chondrocyte Inflammation[J]. Science and Technology of Food Industry, 2022, 43(15): 165−174. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2021100051
Citation: LIU Hengge, XU Xinxing, BAI Fan, et al. Optimization of Sturgeon Cartilage Products and Its Regulation on Human Chondrocyte Inflammation[J]. Science and Technology of Food Industry, 2022, 43(15): 165−174. (in Chinese with English abstract). doi:  10.13386/j.issn1002-0306.2021100051

鲟鱼软骨制品的工艺优化及其对人软骨细胞炎症的调控

doi: 10.13386/j.issn1002-0306.2021100051
基金项目: 国家重点研发计划项目(2018YFD0700901);财政部和农业农村部:国家现代农业产业技术体系(CARS-46)。
详细信息
    作者简介:

    刘恒阁(1997−),男,硕士研究生,研究方向:水产品高值化利用,E-mail:vulpesyolo0819@163.com

    通讯作者:

    刘康(1988−),男,博士,研究方向:水产品高值化利用,E-mail:liukang1888@ouc.edu.cn

    赵元晖(1979−),男,博士,副教授,研究方向:水产品高值化利用,E-mail:zhaoyuanhui@ouc.edu.cn

  • 中图分类号: TS254.9

Optimization of Sturgeon Cartilage Products and Its Regulation on Human Chondrocyte Inflammation

  • 摘要: 本文旨在以鲟鱼软骨为原料制得一种鲟鱼软骨制品,并探究其对人软骨细胞炎症的影响。通过高温高压技术对鲟鱼软骨和龙筋进行热液化处理并加工制得了一种鲟鱼软骨制品,通过单因素及正交试验确定了其最佳制备条件,通过制备含药血清来培养人软骨细胞,以MTT、阿尔新兰染色、ELISA等方法考察了鲟鱼软骨制品对白细胞介素1β(Interleukin 1β,IL-1β)诱导细胞炎症模型的影响。结果表明,鲟鱼软骨制品的最佳制备条件为热液化温度120 ℃、热液化时间20 min、熬煮温度80 ℃、熬煮时间30 min,此条件下该产品的感官评分为59.69分,且鲟鱼软骨制品组的含药血清不具备细胞毒性,能够刺激软骨细胞分泌胶原蛋白及蛋白聚糖等细胞外基质。同时,该产品可以显著提升炎症细胞的增殖活力达156%(P<0.05),且可以显著抑制炎症状态下软骨细胞NO、INOS、IL-6、COX-2和PGE2的表达水平(P<0.05)。
  • 图  1  鲟鱼软骨制品的制备工艺流程

    Figure  1.  Preparation process of sturgeon cartilage products

    图  2  不同热液化条件对鲟鱼软骨制品感官品质的影响

    Figure  2.  Effects of different thermal liquefaction conditions on sensory quality of sturgeon cartilage product

    注:(a)为热液化时间;(b)为热液化温度。

    图  3  不同熬煮条件对鲟鱼软骨制品感官品质的影响

    Figure  3.  Effects of different boiling conditions on sensory quality of sturgeon cartilage product

    注:(a)为熬煮时间;(b)为熬煮温度。

    图  4  鲟鱼软骨制品外观图

    Figure  4.  Appearance of sturgeon cartilage product

    注:(a)、(b)为鲟鱼软骨制品外观侧视图和俯视图。

    图  5  含药血清中羟脯氨酸含量

    Figure  5.  Hydroxyproline content in drug-containing serum

    注:不同的小写字母表示组间差异显著(P<0.05);图7图8同。

    图  6  软骨细胞阿尔新蓝染色

    Figure  6.  Alcian blue staining of chondrocytes

    注:1:未造模组;2:空白对照组;3:空白血清组; 4: 阳性对照组;5~7:鲟鱼软骨制品血清低、中、高浓度组。

    图  7  不同组含药血清对软骨细胞活力的影响

    Figure  7.  Effect of different groups of drug-containing serum on chondrocyte viability

    图  8  不同组含药血清对软骨细胞培养液中细胞因子表达水平的影响

    Figure  8.  Effect of different groups of drug-containing serum on cytokine expression in chondrocyte culture medium

    注:(A)、(B)、(C)分别为INOS和NO、IL-8和IL-6、 COX-2和PGE2的表达水平。

    表  1  正交试验因素水平设计

    Table  1.   Factor and level of orthogonal experimental

    水平因素
    热液化温度(℃)热液化时间(min)熬煮温度(℃)熬煮时间(min)
    1110157020
    2120208030
    3130259040
    下载: 导出CSV

    表  2  感官评定标准

    Table  2.   Sensory evaluation standard

    项目总分(分)
    色泽性状气味、滋味
    浅棕黄色,色泽透亮均匀质地均匀透明,无分层香味浓郁协调,无腥味,无苦涩感,细腻顺滑46~60
    浅棕黄色,色泽略暗均匀质地均匀,略透明,无分层香味稍淡,无腥味,略涩,口感顺滑31~45
    棕黄色,色泽暗淡质地略不均匀,有絮状物漂浮无香味,略带腥味,偏甜或偏酸,略涩,口感略微下降16~30
    深棕色,色泽较黑不均匀质地不均匀,有絮状沉淀难以接受的臭味,腥味重,过甜或过酸,有涩味,口感难以接受0~15
    下载: 导出CSV

    表  3  各单糖成分的线性回归方程和范围

    Table  3.   The linear equations and linear ranges of each monosaccharide

    单糖成分线性回归方程决定系数(R2线性范围(μg/mL)
    甘露糖y=26550x−154420.99961.50~650.00
    氨基葡萄糖y=18445x−150040.99911.50~650.00
    葡萄糖醛酸y=32008x+1233540.99941.50~650.00
    氨基半乳糖y=18664x+121160.99971.50~650.00
    葡萄糖y=18008x−4876600.99921.50~650.00
    半乳糖y=38562x−66240.99941.50~650.00
    下载: 导出CSV

    表  4  正交试验结果

    Table  4.   Results of orthogonal experiment

    实验号A热液化温度B热液化时间C熬煮温度D熬煮时间感官评分(分)
    1111152.86
    2122258.11
    3133355.72
    4212359.19
    5223159.17
    6231258.51
    7313258.33
    8321357.78
    9332157.12
    K1166.69170.38169.15169.15
    K2176.87175.06174.42174.95
    K3173.23171.35173.22172.69
    k155.5656.7956.3856.38
    k258.9658.3558.1458.32
    k357.7457.1257.7457.56
    R3.401.561.761.94
    下载: 导出CSV

    表  5  方差分析结果

    Table  5.   Results of variance analysis

    方差来源自由度F显著性
    热液化温度23.58*
    热液化时间20.43
    熬煮温度20.56
    熬煮时间20.64
    注:*表示显著,P<0.05。
    下载: 导出CSV

    表  6  鲟鱼软骨制品理化指标检测结果

    Table  6.   Results of physical and chemical indexes of sturgeon cartilage product

    项目结果定量限限值说明
    甲基汞(mg/kg)未检出0.025≤1.0符合
    多氯联苯(PCBs)(mg/kg)未检出≤0.5符合
    N-二甲基硝胺(μg/kg)未检出1.0≤4.0符合
    铅(Pb)(mg/kg)未检出0.04≤1.0符合
    铬(Cr)(mg/kg)未检出0.1≤2.0符合
    镉(Cd)(mg/kg)未检出0.003≤0.1符合
    苯甲酸(g/kg)未检出0.010不得检出符合
    无机砷(mg/kg)未检出0.08≤0.1符合
    下载: 导出CSV

    表  7  鲟鱼软骨制品微生物指标检测结果

    Table  7.   Results of microbiological index detection of sturgeon cartilage product

    项目结果ncmM说明
    沙门氏菌/25 g未检出500符合
    金黄色葡萄糖球菌(CFU/g)<10511001000符合
    副溶血性弧菌(MPN/g)<0.3511001000符合
    注:n:同一批次产品应采集的样品件数;c:最大可允许超出m值的样品数;m:微生物指标可接受水平的限量值;M:微生物指标的最高安全限量值。
    下载: 导出CSV

    表  8  鲟鱼软骨制品主要营养成分检测结果

    Table  8.   Results of main nutrients of sturgeon cartilage product

    项目(g/100 g)方法结果检出限
    胶原蛋白羟脯氨酸含量换算2.200.1
    硫酸软骨素GB/T 20365-20062.820.1
    氨基葡萄糖GB/T 20365-20061.170.1
    注:羟脯氨酸含量换算方法,参考陆剑锋等[20]的方法。
    下载: 导出CSV

    表  9  含药血清中单糖成分分析

    Table  9.   Analysis of monosaccharide composition in drug-containing serum

    组别摩尔浓度比率(%)
    甘露糖(%)氨基葡萄糖(%)葡萄糖醛酸(%)氨基半乳糖(%)葡萄糖(%)半乳糖(%)
    空白对照组27.56±0.12a27.66±0.04c0.99±0.16a0.58±0.04b28.40±0.74c14.80±1.42b
    阳性对照组24.44±0.24bc31.83±1.24a0.16±0.02c0.62±0.14a31.05±0.86b11.90±0.24cd
    产品低浓度组26.30±0.09ab30.04±0.84b0.08±0.06d0.57±0.27b27.34±1.24d15.67±0.06ab
    产品中浓度组25.15±0.14b32.26±1.66a0.08±0.08d0.59±0.12b31.08±1.08b10.84±1.04d
    产品高浓度组23.43±0.60c30.01±0.57b0.22±0.04b0.64±0.02a33.05±0.26a12.65±0.84c
    注:同列不同小写字母表示组间差异显著(P<0.05)。
    下载: 导出CSV
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  • 收稿日期:  2021-10-12
  • 网络出版日期:  2022-06-30
  • 刊出日期:  2022-08-03

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