陶素素,彭志兰,陈晖,等.方格星虫酶解超滤组分对顺铂所致HK-2人肾小管上皮细胞损伤的保护作用研究[J].中国海洋药物,2024,43(2):41-52.
方格星虫酶解超滤组分对顺铂所致HK-2人肾小管上皮细胞损伤的保护作用研究
Protective effect of ultrafiltration fractions of Sipunculus nudus enzymatic hydrolysate on cisplatin-induced HK-2 human renal tubular epithelial cell damages
投稿时间:2022-10-17  修订日期:2023-04-10
DOI:
中文关键词:  方格星虫  超滤组分  NF-κB信号通路  顺铂  人肾小管上皮细胞
English Keywords:Sipunculus nudus  Enzymolysis products  NF-κB signaling pathway  Cisplatin  HK-2
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作者单位E-mail
陶素素 广东医科大学 t463262157@163.com 
彭志兰 广东医科大学海洋医药研究院 amypeng037@163.com 
陈晖 自然资源部第三海洋研究所 chenhui@tio.org.cn 
李茹 广东医科大学海洋医药研究院 1135743908@qq.com 
刘华锋 广东医科大学附属医院肾病研究所 hf-liu@263.net 
杨陈 广东医科大学附属医院肾病研究所 yangchen307@126.com 
戚怡* 广东医科大学海洋医药研究院 qiyi7272@gdmu.edu.cn 
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中文摘要:
      目的 探究方格星虫酶解产物对顺铂诱导的HK-2人肾小管上皮细胞损伤的改善作用。方法 方格星虫酶解后的离心液,逐级通过50K Da和2.5K Da超滤膜后,获得>50K Da(F-Ⅰ),2.5~50K Da(F-Ⅱ),<2.5K Da(F-Ⅲ)三个超滤组分,分析其营养成分、分子量排布、和氨基酸组成;在顺铂诱导HK-2细胞损伤模型上,评价各超滤组分对顺铂损伤的HK-2细胞活力、细胞凋亡以及细胞内ROS的产生的影响;采用Western Blotting探究其中活性最优F-Ⅰ超滤组分对NF-κB信号通路的影响。结果 营养成分和分子量排布分析发现F-Ⅰ超滤组分中总糖所占的比例最高,以干基计约占48.59%,逐级超滤可以提高超滤组分中多肽的含量,F-Ⅲ组分主要为<2.5K Da的小分子肽。氨基酸组成分析发现三个超滤组分氨基酸组成齐全,疏水性氨基酸含量高。细胞实验结果表明三个超滤组分均能显著提高顺铂损伤后的HK-2细胞的细胞活力,抑制细胞凋亡和减少细胞内活性氧ROS的累积。其中F-Ⅰ组分缓解顺铂对HK-2细胞损伤的效果最好,能抑制NF-κB信号通路p65和Iκb-α蛋白的磷酸化。结论 方格星虫的三个酶解超滤组分均对顺铂顺伤后的HK-2细胞具有保护作用。相较于其他两个组分,含有较高多糖的F-Ⅰ组分具有更好的改善作用,可能机制与F-Ⅰ组分能更好抑制ROS的积累、细胞凋亡以及NF-κB信号通路有关。
English Summary:
      This study was aimed to investigate the protective effect of ultrafiltration fractions of Sipunculus nudus enzymatic hydrolysate on cisplatin-induced HK-2 cell damages. Firstly, Sipunculus nudus was hydrolyzed by trypsase, then gradually ultra-filtrated by 50K Da and 2.5K Da membrane with molecular weight cutoffs to separate three ultrafiltration fractions, including >50K Da (F-Ⅰ), 2.5~50K Da (F-Ⅱ) and <2.5K Da (F-Ⅲ). Subsequently, the nutritional ingredients and the amino acid composition of three fractions were further analysed. Finally, the ameliorative effects of three fractions were tested in a cisplatin-damaged renal toxicity model by evaluating its cell viability, cell apoptosis and intracellular ROS production level in HK-2 cells. The nutrient composition analysis showed that the proportion of polysaccharides in F-Ⅰ ultrafiltration fraction was the highest, about 48.59%, and the content of peptides in ultrafiltration fractions could be increased by gradually ultrafiltration, the main component of F-Ⅲ was peptides with molecular weight below <2.5K Da. Three ultrafiltration fractions had complete amino acid composition and high hydrophobic amino acid content. The results of cell experiment showed that three fractions could significantly improve the cell viability, inhibit cell apoptosis and reduce the accumulation of intracellular reactive oxygen species (ROS) in HK-2 cells after cisplatin exposure. The effect of F-Ⅰ fraction is significantly better than that of other fractions, which possibly related to better regulating ROS accumulation, cell apoptosis and NF-κB signaling pathway with its higher polysaccharides property.
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