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
陶素素 1彭志兰 2陈晖 3李茹 2刘华锋 4杨陈 4戚怡2
作者信息
- 1. 广东医科大学药学院,广东湛江 524000
- 2. 广东医科大学药学院,广东湛江 524000;广东医科大学海洋医药研究院,广东湛江 524000
- 3. 自然资源部第三海洋研究所,福建厦门 361000
- 4. 广东医科大学药学院,广东湛江 524000;广东医科大学附属医院肾病研究所,广东湛江 524000
- 折叠
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 50 kDa 和 2.5 kDa超滤膜后,获得＞50 kDa(F-Ⅰ),2.5 kDa～50 kDa(F-Ⅱ),＜2.5 kDa(F-Ⅲ)3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＜2.5 kDa的小分子肽.氨基酸组成分析发现3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3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IKb-α蛋白的磷酸化.结论 方格星虫的3个酶解超滤组分均对顺铂损伤后的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.
Abstract
Objective To investigate the protective effect of ultrafiltration fractions of Sipunculus nudus enzymatic hydrolysate on cisplatin-induced HK-2 cell damages.Methods Sipunculus nudus was hydrolyzed by trypsase,then gradually ultra-filtrated by 50 kDa and 2.5 kDa membrane with molecular weight cutoffs to separate three ultrafiltration fractions,including＞50 kDa(F-Ⅰ),2.5 kDa-50 kDa(F-Ⅱ)and＜2.5 kDa(F-Ⅲ).The nutritional ingredients and the amino acid composition of three fractions were further analysed.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 effect of the most active F-Ⅰ ultrafiltration component on the NF-κB signaling pathway was investigated by Western blotting.Results 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＜2.5 kDa.Three ultrafiltration fractions had complete amino acid composition and high hydrophobic amino acid content.Cell experiment showed that three fractions significantly improved the cell viability of HK-2 cells,inhibited its cell apoptosis and reduced the accumulation of intracellular reactive oxygen species(ROS)in HK-2 cells after cisplatin exposure.F-Ⅰ had the best effect on cisplatin-induced damages of HK-2 cells,as well as inhibited the phosphorylation of p65 and Iκb-α proteins in the NF-κB signaling pathway.Conclusion The effect of F-Ⅰfraction on cisplatin-induced cell damages was significantly better than that of other fractions,which possibly related to better regulating ROS accumulation,cell apoptosis and NF-κB signaling pathway in HK-2 cells.
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Key words
Sipunculus nudus/enzymolysis products/NF-κB signaling pathway/cisplatin/human renal tubular epithelial cell引用本文复制引用
基金项目
南海海洋生物制品产业化关键技术研发与创新示范平台项目(2019A01005)
广东省医学科研项目(A2021190)
出版年
2024
NETL
NSTL
万方数据