双斑东方鲀鱼皮胶原多肽对UVB辐射小鼠成纤维细胞的光损伤修复功效

    Photoprotective effect of collagen peptides from the skin of Takifugu bimaculatus on UVB-induced damage in mouse fibroblasts

    • 摘要:
      背景 双斑东方鲀(Takifugu bimaculatus)是福建省养殖河鲀的主要品种之一。近年来,双斑东方鲀养殖产量的显著提高对其鱼皮等加工副产物的高值化利用提出了更加迫切的要求。
      目的 长期暴露于紫外线B(UVB)导致皮肤炎症、真皮层胶原蛋白减少等皮肤光损伤现象。本研究以UVB辐照小鼠成纤维细胞L929为光损伤模型,探讨低分子量双斑东方鲀鱼皮胶原多肽(TBCP-L)对UVB暴露L929细胞的修复作用。
      方法 本研究首先通过体外模型验证双斑东方鲀鱼皮胶原多肽(TBCP)的安全性,随后通过超滤获得了分子量小于3 kD的TBCP-L,利用细胞活性试验探究TBCP-L对UVB辐照L929细胞的预防和修复功效。荧光定量聚合酶链式反应(qPCR)检测UVB暴露L929细胞促炎因子及基质金属蛋白酶(MMPs)的信使核糖核酸(mRNA)表达水平。通过细胞划痕试验研究TBCP-L对伤口创面的修复功效。
      结果 EpiskinTM 3D皮肤模型和牛角膜试验结果表明,1.5 g/mL TBCP溶液对皮肤及眼睛无刺激性。分子量分布测定结果表明,TBCP-L的重均分子量为2352 Da,其中低于2 200 Da的占比达到80.2%。2 mg/mL TBCP-L对L929细胞无毒性作用;对UVB辐射L929细胞无预防作用,但有良好的修复活性。qPCR检测结果表明,TBCP-L可显著抑制UVB刺激后L929中促炎因子白细胞介素-1β、白细胞介素-6、肿瘤坏死因子-α和环氧化酶-2以及MMP2MMP3MMP9的上调表达。同时孵育500 μg/mL TBCP-L 96 h可显著加快L929细胞划痕的修复速度,迁移率达到37.8%±3.3%。
      结论 TBCP-L可能通过抑制炎症反应和胶原蛋白流失加快对UVB辐照的L929细胞的修复,且对皮肤及眼无刺激性,是抗老化和晒后修复化妆品原料的潜在候选物质。
      意义 本研究旨在深入探讨TBCP-L作为缓解光老化和晒后修复化妆品原料的潜在应用潜力,为其成为该领域的有效候选物质提供坚实的理论支撑。同时,对于推动河鲀资源的高值化利用及产业的可持续发展具有深远的实践意义。

       

      Abstract:
      Background Takifugu bimaculatus is one of the main species of cultured puffer fish in Fujian Province. The significant increase in its production in recent years has led to an increasingly urgent need for high-value utilization of processing by-products such as its skin.
      Objective Long-term exposure to ultraviolet B (UVB) radiation leads to skin inflammation and a reduction in dermal collagen, among other photodamage phenomena. This study utilized UVB-irradiated mouse fibroblasts as a model for photodamage to investigate the reparative effects of low molecular weight collagen peptides derived from T. bimaculatus fish skin (TBCP-L) on UVB-exposed L929 cells.
      Methods The study first verified the safety of TBCP through in vitro models. Subsequently, TBCP-L with a molecular weight less than 3 kD was obtained through ultrafiltration. Cell viability assays were conducted to evaluate the preventive and reparative effects of TBCP-L on UVB-irradiated L929 cells. Real-time quantitative polymerase chain reaction (qPCR) was used to detect the messenger ribonucleic acid (mRNA) expression levels of inflammatory cytokines and matrix metalloproteinases (MMPs) in UVB-exposed L929 cells. The wound healing effect of TBCP-L was studied through a cell scratch assay.
      Results EpiskinTM 3D models and bovine corneal experiments indicated that 1.5 g/mL TBCP solution was non-irritating to skin and eyes. The molecular weight distribution results indicated that the weight-average molecular weight of TBCP-L was 2352 Da, with 80.2% of the components having a molecular weight below 2200 Da. A 2 mg/mL TBCP-L concentration was non-toxic to L929 cells and did not prevent UVB radiation damage but exhibited significant reparative activity. qPCR results indicated that TBCP-L could significantly inhibit the upregulated expression of inflammatory cytokines such as interleukin-1β, interleukin-6, tumor necrosis factor-α and cyclooxygenase-2 and MMP2, MMP3, and MMP9 in L929 cells following UVB stimulation. Additionally, incubation with 500 μg/mL TBCP-L for 96 h significantly accelerated the scratch repair rate of L929 cells, with a migration rate reaching 37.8%±3.3%.
      Conclusion In conclusion, TBCP-L may accelerate the repair of UVB-irradiated L929 cells by inhibiting inflammatory responses and collagen loss, and it is non-irritating to the skin and eyes. It is a potential candidate for anti-aging and post-sun exposure repair cosmetic ingredients.
      Significance  This study aims to investigate the potential of TBCP-L as a cosmetic ingredient for mitigating photoaging and post-sun exposure repair, providing robust theoretical support for its efficacy. Additionally, it holds significant practical implications for the high-value utilization of puffer fish resources and the sustainable development of the industry.

       

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