食品与发酵工业 >

2021 , Vol. 47 >Issue 21: 132 - 139

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.026921

研究报告

草苁蓉多糖对过氧化氢致HL-7702细胞损伤的保护作用

  • 金爱花 ,
  • 常慧 ,
  • 张钊 ,
  • 葛乃嘉 ,
  • 全吉淑 ,
  • 尹学哲
展开
  • 1(延边大学附属医院,吉林 延吉,133000)
    2(延边大学 医学院,吉林 延吉,133002)
博士研究生(全吉淑教授和尹学哲教授为共同通讯作者,E-mail:quanjs@ybu.edu.cn;yinxz@ybu.edu.cn)

收稿日期: 2021-01-29

  修回日期: 2021-04-03

  网络出版日期: 2021-11-30

基金资助

国家自然科学地区科学基金项目(82060113;81760659)

收起

Protective effect of Boschniakia rossica polysaccharides on H2O2-induced injury of HL-7702 cells

  • JIN Aihua ,
  • CHANG Hui ,
  • ZHANG Zhao ,
  • GE Naijia ,
  • QUAN Jishu ,
  • YIN Xuezhe
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  • 1(Yanbian University Hospital, Yanji 133000, China)
    2(Yanbian University Medical College, Yanji 133002, China)

Received date: 2021-01-29

  Revised date: 2021-04-03

  Online published: 2021-11-30

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摘要

采用草苁蓉多糖(Boschniakia rossica polysaccharides,BRP)对HL-7702细胞进行预保护,再用H2O2处理HL-7702细胞构建肝细胞氧化损伤模型。CCK-8法测定HL-7702细胞存活率,微板法测定HL-7702细胞培养液中乳酸脱氢酶(lactate dehydrogenase,LDH)活性,硫代巴比妥酸法检测HL-7702细胞丙二醛(malondialdehyde,MDA)的生成,Hoechst33342染色法及原位末端标记法观察HL-7702细胞的凋亡情况,蛋白印迹技术检测HL-7702细胞Caspase 3活性片段(C-casp3)、聚腺苷二磷酸核糖聚合酶活性片段[cleaved poly(ADP-ribose) polymerase,C-PARP]、细胞色素c(cytochrome c,Cyto c)、p53、B细胞淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2-associated X protein,Bax)、Bcl-2同源拮抗剂/杀伤剂(Bcl-2 homologous antagonist/killer,Bak)蛋白表达以及丝裂原活化蛋白激酶、核转录因子-κB(nuclear factor-κB,NF-κB)激活情况。结果表明,300 μmol/LH2O2处理HL-7702细胞6 h可使细胞存活率降低58%(P<0.05),表明H2O2能够诱导HL-7702细胞损伤。而50和100 mg/L BRP能够减缓H2O2导致的HL-7702细胞损伤。与模型组比较,BRP高剂量组HL-7702细胞存活率增高30.8%(P<0.05),LDH释放率降低32.8%(P<0.05),细胞MDA生成降低81.0%(P<0.05)。此外,与模型组比较,BRP组HL-7702细胞凋亡明显减少,胞浆Cyto c水平下降(P<0.05),细胞p53、Bak、Bax蛋白表达下调(P<0.05),Bcl-2蛋白表达上调(P<0.05),C-casp3、C-PARP水平降低(P<0.05),磷酸化细胞外信号调节蛋白激酶(phosphorylated extracellular signal-regulated kinase,p-ERK)、磷酸化c-Jun氨基末端激酶(phosphorylated c-Jun N-terminal kinase,p-JNK)以及核NF-κB水平下降(P<0.05)。提示,BRP能够有效保护H2O2诱导的HL-7702细胞损伤,并抑制HL-7702细胞凋亡,此作用可能与BRP对ERK、JNK、NF-κB通路的调控有关。

关键词： 草苁蓉; 多糖; HL-7702细胞; 过氧化氢; 细胞凋亡

本文引用格式

金爱花 , 常慧 , 张钊 , 葛乃嘉 , 全吉淑 , 尹学哲 . 草苁蓉多糖对过氧化氢致HL-7702细胞损伤的保护作用[J]. 食品与发酵工业, 2021 , 47(21) : 132 -139 . DOI: 10.13995/j.cnki.11-1802/ts.026921

Abstract

The protective effect of Boschniakia rossica polysaccharides (BRP) on the injury of HL-7702 cells induced by hydrogen peroxide was investigated. The cellular injury model was established using hydrogen peroxide in HL-7702 cells. HL-7702 cells were pretreated with BRP, and then cell viability of HL-7702 was investigated with the cell counting kit-8 (CCK-8) assay, and the lactate dehydrogenase (LDH) release was tested with the microplate method. Cellular malondialdehyde (MDA) was tested with the thiobarbituric acid method, cell apoptosis was detected by the Hoechst33342 staining and TdT-mediated dUTP nick end labeling (TUNEL) assay. The protein expression of cytochrome c (Cyto c), p53, B cell lymphoma/lewkmia-2 (Bcl-2), Bcl-2-associated X protein (Bax), Bcl-2 homologous antagonist/killer (Bak), cleaved caspase 3 (C-casp3), cleaved poly (ADP-ribose) polymerase (C-PARP) and the activation of mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) were determined with the western blotting method. The results showed that hydrogen peroxide at a concentration of 300 μmol/L decreased the viability of HL-7702 cells by 58% (P<0.05). However, the cell viability of HL-7702 was increased by 30.8% (P<0.05), and the release of LDH was reduced by 32.8% (P<0.05). Besides, the cellular MDA was decreased by 81.0% (P<0.05) in the BRP group as compared with the model group. In addition, BRP inhibited cell apoptosis, reduced the cytoplasmic Cyto c (P<0.05), increased Bcl-2 protein expression (P<0.05), and reduced the expression of p53, Bak, Bax, C-casp3, C-PARP, phosphorylated extracellular signal-regulated kinase (p-ERK), phosphorylated c-Jun N-terminal kinase (p-JNK) as well as the nuclear translocation of NF-κB (P<0.05). It is suggested that BRP could protect HL-7702 cells from hydrogen peroxide-induced oxidative damage, and inhibit the apoptosis of HL-7702 cells through regulating ERK, JNK and NF-κB pathways.

Key words： Boschniakia rossica; polysaccharides; HL-7702 cells; hydrogen peroxide; cell apoptosis

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