桦褐孔菌皂苷的优化提取及其对HepG2细胞脂肪堆积的清除作用

doi:10.13481/j.1671-587x.20180313

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (03): 526-531.doi: 10.13481/j.1671-587x.20180313

桦褐孔菌皂苷的优化提取及其对HepG2细胞脂肪堆积的清除作用

张蕾¹, 张琨², 王德利³, 任小红⁴, 崔娜¹, 刘洋¹, 赵庭勋¹

1. 牡丹江师范学院生命科学与技术学院制药工程系, 黑龙江牡丹江 157011;
2. 吉林大学第二医院研究中心, 吉林长春 130041;
3. 吉林大学生命科学学院生物实验示范中心, 吉林长春 130012;
4. 中国科学院上海药物研究所药物制剂中心, 上海 201210

收稿日期:2017-09-18 出版日期:2018-05-28 发布日期:2018-05-31
通讯作者: 张蕾,讲师(Tel:0453-6511042,E-mail:swxzlz@126.com) E-mail:swxzlz@126.com
作者简介:张蕾(1984-),女,黑龙江省东宁市人,讲师,理学博士,主要从事微生物和生物化学方面的研究。
基金资助:
黑龙江省科技厅重点创新预研项目资助课题（SY2014009）

Optimal extraction of Phaeoporus obliquus saponins and its scavengingeffect on fat accumulation in HepG2 cells

ZHANG Lei¹, ZHANG Kun², WANG Deli³, REN Xiaohong⁴, CUI Na¹, LIU Yang¹, ZHAO Tingxun¹

1. Department of Pharmaceutical Engineering, College of Life Science and Technology, Mudanjiang Normal University, Mudanjiang 157011, China;
2. Research Center, Second Hospital, Jilin University, Changchun 130041, China;
3. Domonstration Center for Biological Experiments, School of Life Sciences, Jilin University, Changchun 130012, China;
4. Center of Pharmaceutical Preparations, Chinese Academy of Sciences, Shanghai Institute of Materia Medica, Shanghai 201210, China

Received:2017-09-18 Online:2018-05-28 Published:2018-05-31

摘要/Abstract

摘要： 目的：探讨桦褐孔菌皂苷清除肝细胞中脂肪堆积的能力，阐明桦褐孔菌皂苷对非酒精性脂肪肝病（NAFLD）的预防效果。方法：MTT法选取桦褐孔菌皂苷对HepG2细胞的安全浓度，将不同浓度（60.000、6.000、0.600、0.060和0.006g·L^-1）桦褐孔菌皂苷作用于HepG2细胞作为给药组，以不给药无细胞的空白孔为对照组，以不给药的HepG2细胞为模型组。MTT法检测各组HepG2细胞存活率，检测各组HepG2细胞中甘油三酯（TG）的清除率，油红O染色检测各组HepG2细胞形态表现。结果：提取桦褐孔菌皂苷最佳条件为1∶15的料液比、温度为30℃、提取时间为30 min、超声功率为50%。与对照组比较，0.060 g·L^-1桦褐孔菌皂苷组细胞存活率降低，TG清除率升高（P<0.05），细胞中脂质相互融合的现象明显减轻；0.600 g·L^-1桦褐孔菌皂苷组细胞存活率降低，TG清除率升高（P<0.01），细胞中脂滴数量明显减少。结论：最佳提取条件下桦褐孔菌皂苷提取率明显升高且具有良好的体外降脂活性，对NAFLD有较好的预防效果。

关键词: 桦褐孔菌, 脂肪堆积, 皂苷, HepG2细胞, 甘油三酯

Abstract: Objective: To investigate the ability of Phaeoporus obliquus saponins in scavenging the fat accumulation of liver cells,and to clarify its effect on the prevention of non-alcoholic fatty liver disease(NAFLD). Methods: The safe concentrations of Phaeoporus obliquus in the HepG2 cells was selected by MTT method. The cells treated with 60.000,6.000,0.600,0.060,and 0.006 g·L^-1 Phaeoporus obliquus saponins were regarded as treatment groups;the blank well without drugs and cells was regarded as control group,and the HepG2 cells without drugs were regarded as model group.The survival rates of HepG2 cells in various groups were detected by MTT method;the clearance rates of triglyceride(TG) in the HepG2 cells in various groups were detected;the morphology of HepG2 cells was detected by oil red O staining. Results: The optimum extraction condition of Phaeoporus obliquus saponins were as followed:the liquid ratio was 1:15,the extraction temperature was 30℃,the extraction time was 30 min, and the ultrasonic power was 50%.Compared with control group,the survival rate of HepG2 cells in 0.060 g·L^-1 Phaeoporus obliquus saponins group was decreased and the clearance rate of TG was increased(P<0.05),and the lipid interfusion in cells was significantly reduced.Compared with control group,the survival rate of HepG2 cells in 0.600 g·L^-1 Phaeoporus obliquus saponins group was decreased and the clearance rate of TG was increased(P<0.01),and the number of lipid drops was significantly reduced. Conclusion: The extraction rate of Phaeoporus obliquus saponins is obviously improved under the optimum extraction conditions and it has good in vitro lipid-lowering activity,and it has better preventive effect on NAFLD.

Key words: Phaeoporus obliquus, saponins, fat accumulation, triglyceride, HepG2 cells

中图分类号:

R282.71

张蕾, 张琨, 王德利, 任小红, 崔娜, 刘洋, 赵庭勋. 桦褐孔菌皂苷的优化提取及其对HepG2细胞脂肪堆积的清除作用[J]. 吉林大学学报(医学版), 2018, 44(03): 526-531.

ZHANG Lei, ZHANG Kun, WANG Deli, REN Xiaohong, CUI Na, LIU Yang, ZHAO Tingxun. Optimal extraction of Phaeoporus obliquus saponins and its scavengingeffect on fat accumulation in HepG2 cells[J]. Journal of Jilin University Medicine Edition, 2018, 44(03): 526-531.

参考文献

[1] Abdelmalek MF,Diehl AM.Nonalcoholic fatty liver disease as a complication of insulin resistance[J]. Med Clin North Am,2007,91(6):1125-1149.
[2] Chow WC,Tai ES,Lian SC,et al.Significant non-alcoholic fatty liver disease is found in non-diabetic, pre-obese Chinese in Singapore[J]. Singapore Med J, 2007,48(8):752-757.
[3] Samuel VT,Liu ZX, Qu X, et al.Mechanism of hepatic insulin resistance in non-alcoholic fatty liver disease[J].J Biol Chem,2004,279(31):32345-32353.
[4] 侯晓琳,周培,杨小林,等.竹节参皂苷Ⅴ对油酸诱导 HepG2细胞脂肪堆积的影响作用[J].海峡药学,2017,29(3):38-40.
[5] Shin Y,Tamai Y,Terazawa M.Chemical constituents of Inonotus boliquus,a new triterpene,21,24-cyclopentalanosta-3β,21,25-triol-8-ene from sclera-tium[J].Wood Res Soc,2001,47(9):313-316.
[6] Cui Y,Kim DS,Park KC.Antioxidant effect of Inonotus obliquus[J].Eth Pha,2005,96(1/2):79-85.
[7] Kim YO,Park HW,Kim JH,et al.Anti-cancer effect and sructural characterization ofendo-polysacchride from cultivated mycelia of Inonotusobliquus[J].Life Sci,2006,79(1):72-80.
[8] Song HS,Lee YJ,Kim SK,et al.Down regulatory effect of AGI-1120(α-glucosidase inhibitor)and Chagamushoom(Inonotus obliquus)on cellular NF-kB acti-vation and their antioxidant activity[J].Kor Jph,2004,35(9):92-97.
[9] 杜秀菊,徐伟,穆红梅,等.桦褐孔菌多糖的药理活性与化学结构研究进展[J].食用菌学报,2012,19(1):100-104.
[10] 李宏.桦褐孔菌多糖的提取工艺优化及其对小鼠免疫功能的影响[J].赤峰学院学报:自然科学版,2012,28(2):128-130.
[11] 计红芳,张令文,孙科祥,等.玉米须总皂苷的微波辅助提取及抑菌活性研究[J].中国食品添加剂试验研究,2012,11(3):103-107.
[12] 陈勋,于海宁,唐德松,等.苦瓜皂甙快速分离纯化方法研究[J].食品科学,2004,25(2):114-117.
[13] Cousin SP,Hugl SR,Wrede CE,et al. Free fatty acid -induced inhibition of glucose and insulin-like growth factor I-induced deoxyribonucleic acid synthesis in the pancreatic beta-cell line INS-1[J].Endocrinology,2001,142(1):229-240.
[14] 陈静,祁东利,张春风,等.人参皂苷 Rb1 对油酸诱导 HepG2 细胞脂肪堆积的影响作用[J].中国野生植物资源,2013,32(2):20-23.
[15] 孙丽娜,杨中林.川续断皂苷对油酸诱导细胞脂肪堆积的干预作用[J].药学与临床研究,2014,22(4):326-328.
[16] 丁晓东,范建高.非酒精性肝脂肪变性和脂肪性肝炎的发生机制[J].现代消化及介入诊疗,2009,14(4):237-242.
[17] 李甜,廖茂梁,郑雅楠,等.泽泻汤对油酸诱导HepG2细胞脂肪堆积的抑制作用[J].现在药物与临床,2017,32(5):772-775.
[18] Sonia MN,Rexford SA.Metabolic basis of obesity[J].Springer New York,2011(10):219-227.
[19] 杨林辉,陈东风.油酸诱导培养肝细胞脂肪变性模型的建立[J].重庆医学,2007,36(8):698-700.
[20] 魏颖,鲁军,刘艳,等.玉米低聚肽对Hep G2细胞脂肪堆积和抗氧化活性的影响[J].食品科技,2013,38(10):190-193.
[21] 李明辉,刘洋,陈冲,等.自体脂肪源性干细胞联合无菌生物护创膜对慢性创面愈合的影响[J].解放军医学杂志,2016,41(12):1025-1030.

桦褐孔菌皂苷的优化提取及其对HepG2细胞脂肪堆积的清除作用

Optimal extraction of Phaeoporus obliquus saponins and its scavengingeffect on fat accumulation in HepG2 cells

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