Study on the protective effect and mechanism of seabuckthorn polysaccharides on insulin-resistant HepG2 cells from oxidative stress
Objective:The protective effect and mechanism of seabuckthorn polysaccharides on insulin-resistant HepG2 cells against oxidative stress was investigated.Methods:The seabuckthorn polysaccharide was extracted by water extraction and alcohol precipitation method, and the polysaccharide content was determined by the phenol sulfuric acid method. The CCK-8 method was used to determine the effects of different concentrations of sea-buckthorn polysaccharides on the viability of HepG2 cells. HepG2 cells were induced with a medium containing 5×10-8 mol/L insulin for 24 hours to establish a HepG2 cell insulin resistance model. A kit was used to determine glucose consumption and relative glycogen content, and to determine the content of superoxide dismutase (SOD) and malondialdehyde (MDA). Western Blot method was used to determine the expression of oxidative stress-related proteins.Results:The content of sea-buckthorn polysaccharide was 88.46%. When the concentration of sea-buckthorn polysaccharide reached 800 μg/mL, the cell viability decreased significantly (P＜0.05). The subsequent safe dose was 400 μg/mL. Glucose consumption and relative glycogen content after seabuckthorn polysaccharide treatment were significantly increased (P＜0.05), SOD level reached (79.31±2.16) U/mg, MDA level reached (2.15±0.12) nmol/mg. After seabuckthorn polysaccharide treatment, the expression level of Nrf2 and HO-1 was significantly increased (P＜0.05), and the expression level of Keap1 was significantly reduced (P＜0.05).Conclusion:Hippophae rhamnoides polysaccharide can improve the abnormal glucose metabolism and oxidative stress level of insulin resistance cell model, and can improve insulin resistance by regulating the Nrf2/Keap1/HO-1 pathway.
Qiu-dan, WANG; Kai-di, ZHAO; and Chang-qing, LIN
"Study on the protective effect and mechanism of seabuckthorn polysaccharides on insulin-resistant HepG2 cells from oxidative stress,"
Food and Machinery: Vol. 38:
3, Article 27.
Available at: https://www.ifoodmm.cn/journal/vol38/iss3/27
 刘雅娜,包晓玮,王娟,等.沙棘多糖抗运动性疲劳及抗氧化作用的研究[J].食品工业科技,2021,42(10):321-326.LIU Ya-na,BAO Xiao-wei,WANG Juan,et al.Research on anti-exercise fatigue and antioxidant effects of seabuckthorn polysaccharides[J].Science and Technology of Food Industry,2021,42(10):321-326.
 李静,樊佳妮,张龄芷,等.活性植物多糖在食品领域的应用综述[J].上海师范大学学报(自然科学版),2021,50(2):162-169.LI Jing,FAN Jia-ni,ZHANG Ling-zhi,et al.Overview of the application of active plant polysaccharides in the field of food[J].Journal of Shanghai Normal University(Natural Science Edition),2021,50(2):162-169.
 杨玉红,郝慧敏.药用植物多糖的生物学功能及应用研究进展[J].生物学教学,2013,38(1):6-8.YANG Yu-hong,HAO Hui-min.Research progress on the biological function and application of medicinal plant polysaccharides[J].Biology Teaching,2013,38(1):6-8.
 武美馥.响应面优化闪式提取沙棘多糖工艺及其生物活性的研究[D].吉林:吉林化工学院,2019:24-26.WU Mei-fu.Study on the flash extraction process and biological activity of Hippophae rhamnoides polysaccharide by response surface optimization[D].Jilin:Jilin Institute of Chemical Technology,2019:24-26.
 吕志琴,童军茂,陈国刚.沙棘多糖提取纯化工艺[J].农产品加工,2014(9):44-45.LU Zhi-qin,TONG Jun-mao,CHEN Guo-gang.Extraction and purification process of seabuckthorn polysaccharides[J].Agricultural Products Processing,2014(9):44-45.
 刘明杰.沙棘多糖的分离纯化及其抗运动性疲劳作用[J].食品安全质量检测学报,2021,12(10):4 009-4 013.LIU Ming-jie.Isolation and purification of sea buckthorn polysaccharides and its anti-exercise fatigue effect[J].Journal of Food Safety and Quality Inspection,2021,12(10):4 009-4 013.
 魏晨业,包晓玮,王娟,等.沙棘多糖分离纯化及抗氧化活性[J].食品科学,2021,42(4):227-232.WEI Chen-ye,BAO Xiao-wei,WANG Juan,et al.Isolation,purification and antioxidant activity of seabuckthorn polysaccharides[J].Food Science,2021,42(4):227-232.
 王毅兵.沙棘多糖的响应面提取及抗炎作用研究[J].食品工业,2013,34(10):22-24.WANG Yi-bing.Response surface extraction and anti-inflammatory effects of seabuckthorn polysaccharides[J].Food Industry,2013,34(10):22-24.
 GUNDMI S,MAIYA A G,BHAT A K,et al.Hand dysfunction in type 2 diabetes mellitus:Systematic review with meta-analysis[J].Annal Phys Rehabiliati Med,2018,61(2):99-104.
 FENG W,LIU Y,FEI F,et al.Improvement of high-glucose and insulin resistance of chromium malate in 3T3-L1 adipocytes by glucose uptake and insulin sensitivity signaling pathways and its mechanism[J].RSC Adv,2019,9(1):114-127.
 SAEEDI P,PETERSOHN I,SALPEA P,et al.Global and regional diabetes prevalence estimates for 2019 and projections for 2030 and 2045:Results from the international diabetes federation diabetes atlas[J].Diabetes Res Clin Pract,2019,157:107843.
 励丽.扒一扒降糖药的副作用[J].大众健康,2021(4):58-59.LI Li.Side effects of antidiabetic drugs for Pa Yipa[J].Public Health,2021(4):58-59.
 姚兰,蒋成霞,郭艳艳,等.沙棘多糖抑制PERK/ATF4/CHOP通路缓解糖尿病大鼠胰岛素抵抗和肝肾功能损伤[J].中国临床解剖学杂志,2021,39(2):187-191.YAO Lan,JIANG Cheng-xia,GUO Yan-yan,et al.Hippophae rhamnoides polysaccharide inhibits PERK/ATF4/CHOP pathway to relieve insulin resistance and liver and kidney damage in diabetic rats[J].Chinese Journal of Clinical Anatomy,2021,39(2):187-191.
 祝敏,展俊岭,杨洁,等.沙棘叶多糖提取方法及生物活性研究现状[J].化工时刊,2018,32(12):36-38.ZHU Min,ZHAN Jun-ling,YANG Jie,et al.Research status of extraction methods and biological activity ofseabuckthorn leaves polysaccharides[J].Chemical Industry Times,2018,32(12):36-38.
 罗春萍,陆友利,王星星.苯酚—硫酸法快速测定多糖方法的优化[J].化工管理,2021(3):90-91,94.LUO Chun-ping,LU You-li,WANG Xing-xing.Optimization of phenol-sulfuric acid method for rapid determination of polysaccharides[J].Chemical Industry Management,2021(3):90-91,94.
 刘迪迪,邱军强,程翠林,等.HepG2细胞胰岛素抵抗模型建立影响因素研究[J].食品与药品,2018,20(1):1-6.LIU Di-di,QIU Jun-qiang,CHENG Cui-lin,et al.Study on the influencing factors of the establishment of HepG2 cell insulin resistance model[J].Food and Drugs,2018,20(1):1-6.
 周玉荣.桃叶珊瑚苷对胰岛素抵抗的改善作用机制研究[D].大连:大连理工大学,2021:31-32.ZHOU Yu-rong.Study on the mechanism of aucubin in improving insulin resistance[D].Dalian:Dalian University of Technology,2021:31-32.
 CHEN J C,LI L,ZHANG X,et al.Structural characterization of polysaccharide from Centipeda minima and its hypoglycemic activity through alleviating insulin resistance of hepatic HepG2 cells[J].Journal of Functional Foods,2021,82:104478.
 刘亚萍,李雨蒙,王梦,等.菊粉复配灵芝多糖对HepG2细胞胰岛素抵抗的调节作用[J].中国食品添加剂,2018(12):116-121.LIU Ya-ping,LI Yu-meng,WANG Meng,et al.The modulating effect of inulin compounded with Ganoderma lucidum polysaccharide on insulin resistance of HepG2 cells[J].China Food Additives,2018(12):116-121.
 ZHU Y,BAI J,ZHOU Y,et al.Water-soluble and alkali-soluble polysaccharides from bitter melon inhibited lipid accumulation in HepG2 cells and Caenorhabditis elegans[J].International Journal of Biological Macromolecules,2021,166:155-165.
 WEI E,YANG R,ZHAO H,et al.Microwave-assisted extraction releases the antioxidant polysaccharides from seabuckthorn(Hippophae rhamnoides L.)berries[J].International Journal of Biological Macromolecules,2019,123:280-290.
 WASIK U,MILKIEWICZ M,KEMPINSKA-PODHORODECKA A,et al.Protection against oxidative stress mediated by the Nrf2/Keap1 axis is impaired in primary biliary cholangitis[J].Scientific Reports,2017,7:44769.