海洋深层水与褐藻糖胶联合用药对IR-HepG2细胞氧化应激水平的影响  

Combination treatment of DSW and FPS protected against oxidative stress in IR-HepG2 cells

作　　者：何珊[1] 彭维兵[2,3] 周洪雷 HE Shan;PENG Wei-bing;ZHOU Hong-lei(School of Pharmaceutical Sciences,Shandong University of Traditional Chinese Medicine,Jinan 250355,China;Biology Institute of Shandong Academy of Sciences,Engineering I.aboratory for Biological Testing Technology of Shandong Province,Key I.aboratory for Biosensor of Shandong Province,Key Laboratory for Drug Screening Technology of Shandong Academy of Sciences,Jinan 250014,China;Qilu University of Technology（Shandong Academy of Sciences）,Jinan 250014,China)

机构地区：[1]山东中医药大学药学院,山东济南250355 [2]山东省科学院生物研究所山东省生物检测技术工程实验室山东省生物传感器重点实验室山东省科学院药物筛选技术重点实验室,山东济南250014 [3]齐鲁工业大学(山东省科学院),山东济南250014

出　　处：《中国海洋药物》2018年第3期32-40,共9页Chinese Journal of Marine Drugs

基　　金：山东省高等学校科技计划项目(J16LM07);山东省自然科学基金项目(ZR2015HM080);一流学科中药药效物质发现与评价研究项目(220311)资助

摘　　要：目的探讨海洋深层水(deep-sea water,DSW)与褐藻糖胶(fucoidan,FPS)联合用药对胰岛素抵抗(insulin resistance,IR)HepG2细胞内氧化应激水平的影响。方法制备FPS和硬度为1 000的DSW,并对其理化性质进行分析。采用MTT法检测DSW与FPS联合用药对HepG2细胞增殖的影响,确定与DSW联合用药的FPS浓度。高浓度葡萄糖(30mmol·L^(-1))诱导建立IR-HepG2细胞模型,同时给予DSW与不同浓度FPS的联合用药,药物孵育24h后,测定肝糖输出量和糖原含量,并以二者为指标确定DSW与FPS的最佳联用剂量;然后对胞内活性氧簇(reactive oxygen species,ROS)和丙二醛(malondialdehyde,MDA)含量及超氧化物歧化酶(Superoxide Dismutase,SOD)、谷胱甘肽过氧化物酶(Glutathione peroxidase,GSH-Px)进行测定,并采用Western blot对C-Jun氨基端激酶(c-jun NH2-terminal kinase,JNK)蛋白的表达及磷酸化程度进行检测,观察DSW与FPS对IR-HepG2氧化应激的影响。结果 DSW与50mg·L^(-1)以内的FPS联合用药不会对细胞增殖产生影响,可以剂量依赖性地降低肝糖输出量,提高糖原水平,其中以DSW与50mg·L^(-1) FPS的联用效果最好,而且其联用效果显著优于单用同剂量的DSW或FPS;DSW与50mg·L^(-1) FPS联用可以显著降低MDA和ROS的水平,提高GSH-Px和SOD的活性,并显著降低JNK的磷酸化程度。结论 DSW与FPS联合用药可降低IR-HepG2细胞内的氧化应激水平,其最佳联用剂量为50mg·L^(-1),其保护作用可能与激活抗氧化酶、抑制脂质过氧化反应及抑制JNK通路有关。Objective To investigate the protective effect of drug combination of deep-sea water（DSW）and fucoidans（FPS）in insulin resistance（IR）-HepG2 cells.Methods DSW of hardness 1 000 and FPS were prepared,then the composition and physicochemical property were elucidated.Thereafter cell proliferation was determined by MTT assay to provide the safe dose for the following experiments.IR cells model was induced by high concentration of glucose（30 mmol·L-1）,at the same time cells were treated with drug combination of DSW and various doses of FPS（5、10、20、50 mg·L-1）for 24 h.The contents of hepatic glucose production and glycogen level in IR-HepG2 were measured;the contents of ROS,MDA,SOD and GSH-Px were observed by spectrophotometric method;the expression and phosphorylation of JNK were detected by Western blot.Results MTT assay showed that the cells were tolerant when they were treated with co-treatment of DSW and FPS with a concentration below 50 mg·L-1.Combination treatment of DSW and FPS could repress hepatic glucose production and increase glycogen level in IR-HepG2 cells in a dose-dependent manner.Moreover,compared with the single use of DSW or FPS,drug combination of DSW and FPS in dosage of 50 mg·L-1 exerted higher potency to decrease hepatic glucose production in IR-HepG2 cells.Compared with the model group,combination treatment of DSW and FPS significantly decreased the levels of MDA and ROS,improved the enzyme activity of SOD and GSH-Px in IR-HepG2 cells.Meanwhile,the phosphorylation of JNK also could be reduced significantly.Conclusion Combination treatment of DSW and FPS could alleviate oxidative stress in IR-HepG2 cells,which might be associated with activation of the antioxidant enzymes,inhibition of lipid peroxidation and JNK pathway.

关 键 词：海洋深层水 褐藻糖胶 IR-HepG2 氧化应激 JNK通路 

分 类 号：R965[医药卫生—药理学]