青钱柳活性成分对IR-HepG2细胞葡萄糖消耗量及α-葡萄糖苷酶活性的影响
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摘要
研究青钱柳多糖(CPP)和青钱柳黄酮(CPF)对胰岛素抵抗的人源肝癌细胞(IR-HepG2)葡萄糖消耗量及α-葡萄糖苷酶活性影响。用高浓度胰岛素诱导培养HepG2细胞,建立IRHepG2细胞模型,将IR-HepG2细胞分为模型组、CPP高、中、低剂量组、CPF高、中、低剂量组和二甲双胍组,各组以相应药物孵育24 h,采用葡萄糖试剂盒测定细胞葡萄糖消耗量(△GC),MTT法测定单位细胞葡萄糖消耗量(△GC/OD);以阿卡波糖为阳性对照,比较不同浓度梯度的CPP与CPF对α-葡萄糖苷酶活性影响。与模型组比较,阴性对照组、二甲双胍组、CPP组与CPF组细胞△GC与△GC/OD显著性升高;不同浓度的CPP与CPF对α-葡萄糖苷酶均有抑制作用。提示青钱柳活性提取物降血糖功效与其提高细胞葡萄糖摄取量,抑制α-葡萄糖苷酶活性有关。
The effects of Cyclocarya paliurus(Batal.) Iljinskaja Extracts on glucose consumption of insulin resistance HepG2 cell and activity of alpha-glucosidase is studied. HepG2 cell with high concentration insulin(10 μg/mL) is induced to establish insulin resistance cell model and divided model cell into Cyclocarya paliurus polysaccharide(CPP),Cyclocarya paliurus flavone(CPF) and dimethylbiguanide groups. After 24 h administration,glucose assay is used fit to measure the gross glucose consumption of cells(△GC), glucose consumption of unite cell(△GC/OD) is detected by MTT assay. The inhibitory activity on alpha-glucosidase of CPP and CPF is evaluated,compared with acarbose,in vitro. Compared with model group,△GC and △GC/OD of IR-HepG2 cell is increased by CPF and CPP significantly. CPP and CPF have inhibitory effect on the activity of alpha-glucosidase. Those results indicat that CPF and CPP have effects on increasing the glucose consumption of cell and decreasing the activity of alpha-glucosidase,thus alleviating hyperglycemia.
The effects of Cyclocarya paliurus(Batal.) Iljinskaja Extracts on glucose consumption of insulin resistance HepG2 cell and activity of alpha-glucosidase is studied. HepG2 cell with high concentration insulin(10 μg/mL) is induced to establish insulin resistance cell model and divided model cell into Cyclocarya paliurus polysaccharide(CPP),Cyclocarya paliurus flavone(CPF) and dimethylbiguanide groups. After 24 h administration,glucose assay is used fit to measure the gross glucose consumption of cells(△GC), glucose consumption of unite cell(△GC/OD) is detected by MTT assay. The inhibitory activity on alpha-glucosidase of CPP and CPF is evaluated,compared with acarbose,in vitro. Compared with model group,△GC and △GC/OD of IR-HepG2 cell is increased by CPF and CPP significantly. CPP and CPF have inhibitory effect on the activity of alpha-glucosidase. Those results indicat that CPF and CPP have effects on increasing the glucose consumption of cell and decreasing the activity of alpha-glucosidase,thus alleviating hyperglycemia.
引文
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[2]XIE Mingyong,XIE Jianhua. Review about the research on Cyclocarya paliurus(Batal.)Iljinskaja[J]. Journal of Food Science and Biotechnology,2008,27(1):113-121.
[3]XIE J H,SHEN M Y,NIE S P,et al. Separation of water-soluble polysaccharides from Cyclocarya paliurus by ultrafiltration process[J]. Carbohydrate Polymers,2014,101(1):479-483.
[4]WANG Q Q,JIANG C H,FANG S Z,et al. Antihyperglycemic,antihyperlipidemic and antioxidant effects of ethanol and aqueous extracts of Cyclocarya paliurus leaves in type 2 diabetic rats[J]. Journal of Ethnopharmacology,2013,150(3):1119-1127.
[5]LI S,LI J,GUAN X L,et al. Hypoglycemic effects and constituents of the barks of Cyclocarya paliurus and their inhibiting activities to glucosidase and glycogen phosphorylase[J]. Fitoterapia,2011,82(7):1081-1085.
[6]XU Mingsheng,SHEN Yonggen,WU Hailong,et al. The hypoglycemic effects of Cyclocarya paliurus(Batal.)Iljinsk water extracts on diabetes mice[J]. Acta Nutrimenta Sinica,2004,26(3):230-234.(in Chinese)
[7]JIANG C H,YAO N,WANG Q Q,et al. Cyclocarya paliurus extract modulates adipokine expression and improves insulin sensitivity by inhibition of inflammation in mice[J]. Journal of Ethnopharmacology,2014,153(2):344-351.
[8]KURIHARA H,FUKAMI H,KUSUMOTO A,et al. Hypoglycemic action of Cyclocarya paliurus(Batal.)Iljinsk in normal and diabetic mice[J]. Biosci Biotechnol Biochem,2003,67(4):877-880.
[9]GU J F,ZHENG Z Y,YUAN J R,et al. Comparison on hypoglycemic and antioxidant activities of the fresh and dried Portulaca oleracea L. in insulin-resistant HepG2 cells and streptozotocin-induced C57BL/6J diabetic mice[J]. Journal of Ethnopharmacology,161(23):214-223.
[10]WILLIAMS J F,OLEFSKY J M. Defective insulin receptor function in down-regulated HepG2 cell[J]. Endocrinology,1990,127(4):1706-1717.
[11]LIN C L,HUANG H C,LIN J K. The aflavins attenuate hepatic lipid accumulation through activating AMPK in human HepG2cells[J]. Journal of Lipid Research,2007,48(11):2334-2343.
[12]LIU Jie,XIANG Yanru,DING Jiayu,et al. Screening the active fraction from Cyclocarya paliurus(Batal.)Iljinskaja which inhibitα-glucosidase and study its effects on hyperglycemia in mice with typeⅡdiabetes mellitus[J]. Science and Technology of Food Industry,2015,36(14):363-369.(in Chinese)
[13]XIE J H,XIE M Y,NIE S P,et al. Chemical composition and antioxidant activities of a water-soluble polysaccharide from Cyclocarya paliurus(Batal.)Iljinskaja[J]. Food Chemistry,2010,119(4):1626-1632.
[14]LAURENTIN A,EDWARDS C A. A microtiter modification of the anthrone-sulfuric acid colorimetric assay for glucose-based carbohydrates[J]. Analytical Biochemistry,2003,315(1):143-145.
[15]XIE J H,DONG C J,NIE S P,et al. Extraction,chemical composition and antioxidant activity of flavonoids from Cyclocarya paliurus(Batal.)Iljinskaja leaves[J]. Food Chemistry,2015,186(2):97-105.
[16]AMIR M,MUJEEB M,KHAN A,et al. Phytochemical analysis and in vitro antioxidant activity of Uncaria gambir[J].International Journal of Green Pharmacy,2012,6(1):67-72.
[17]SU Jin,JIAO Jun,YU Lian,et al. Effects of Dioscorea japonica polysaccharide on glucose consumption capacity and insulin resistance of human lung cancer HepG2 cells[J]. China Pharmacy,2015,26(4):458-460.
[18]方飞.桑叶有效部位改善HepG2胰岛素抵抗及其机制的研究[D].广州:华南理工大学,2012.
[19]LYU Huan,LUO Mingli,FANG Fei,et al. In vitro inhibitory effect of mulberry leaf extract on alpha-glucosidase activity[J].Lishizhen Medicine and Materia Medica Research,2012,23(1):41-42.(in Chinese)
[20]TONG Xing,TANG Qiusong,WU Yufei,et al. Cloning of the gene encodingα-glucosidase from Aspergillus niger and its expression in Pichia pastoris[J]. Acta Microbiologica Sinica,2009,49(2):262-268.(in Chinese)
[21]YANG Wuying,SHANG GUAN Xinchen,XU Mingsheng,et al. Effects of Cyclocarya paliurus flavonoids on activity ofα-glucosidase and blood glucose level in diabetic mice[J]. Acta Nutrimenta Sinica,2007,29(5):507-509.(in Chinese)
[22]JAMES D E,BROWN R,NAVARRO J,et al. Insulin-regulatable tissues express a unique insulin-sensitive glucose transport protein[J]. Nature,1988,333(6169):183-185.
[23]LI Nan,ZHAO Jing,WU Ru,et al. Effects of Cyclocarya paliurus(Batal.)Iljinskaja polysaccharides on lipid metabolism and expressions of PPARα,FAS,and GLUT4 m RNA in hyperlipidemic Rats[J]. Modern Food Science and Technology,2015,31(4):29-35.(in Chinese)
[24]JIN M N,SHI G R,TANG S A,et al. Flavonoids from Tetrastigma obtectum enhancing glucose consumption in insulin-resistance HepG2 cells via activating AMPK[J]. Fitoterapia,2013,90(20):240-246.
[25]MA X L,ZHOU F C,CHEN Y Y,et al. A polysaccharide from Grifola frondosa relieves insulin resistance of HepG2 cell by Akt-GSK-3 pathway[J]. Glycoconjugate Journal,2014,31(5):355-363.