木犀草素和异荭草素对Caco-2细胞中α-葡萄糖苷酶及葡萄糖吸收的影响
|
摘要
目的:研究木犀草素和异荭草素对Caco-2细胞α-葡萄糖苷酶的抑制作用及对葡萄糖吸收的影响。方法:取对数生长期的Caco-2细胞,分别加入不同浓度的木犀草素和异荭草素孵育,CCK-8法检测细胞的存活率,酶标仪检测α-葡萄糖苷酶抑制率,RT-PCR法和Western blot法检测Caco-2细胞钠依赖性葡萄糖转运蛋白1(SGLT1)和葡萄糖转运蛋白2(GLUT2)的mRNA和蛋白表达。结果:100μmol/L木犀草素和异荭草素对α葡萄糖苷酶的抑制作用与阿卡波糖差异无统计学意义(P>0.05);50、100μmol/L木犀草素和100μmol/L异荭草素能显著抑制SGLT1、GLUT2 mRNA及蛋白表达(P<0.05或P<0.01)。结论:木犀草素和异荭草素通过抑制Caco-2细胞的α-葡萄糖苷酶和SGLT1、GLUT2的mRNA和蛋白表达来延缓对碳水化合物水解和影响葡萄糖吸收,具有潜在的降血糖作用。在相同浓度下,木犀草素的效果要优于异荭草素。
Objective:To investigate the effects of luteolin and isoorientin on α-glucosidase enzyme activity and glucose absorption in Caco-2 Cells.Methods:Caco-2 cells in logarithmic phase were clutivated with different concentrations of luteolin and isoorientin.The survival rate of cells was determined by CCK-8 method,and the inhibition rate of α-glucosidase was determined by microplate reader.The mRNA and protein expressions of SGLT-1 and GLUT-2 in Caco-2 cells were detected by RT-PCR and Western blot.Results:The inhibitory effect of 1 μmol/L Luteolin and isoorientin on α-glucosidase was not statistically significant with that of acarbose(P>0.05);50 and 100 μmol/L Luteolin and 100 μmol/L isoorientin SGLT1,GLUT2 mRNA and protein expressions(P<0.05 or P<0.01).Conclusion:Luteolin and isoorientin can delay the hydrolysis of carbohydrates and inhibit the glucose absorption by inhibiting the α-glucosidase enzyme and the mRNA and protein expressions of SGLT1 and GLUT2 in Caco-2 cells.They both have potential hypoglycemic effect.At the same concentration,luteolin is superior to isoorientin.
Objective:To investigate the effects of luteolin and isoorientin on α-glucosidase enzyme activity and glucose absorption in Caco-2 Cells.Methods:Caco-2 cells in logarithmic phase were clutivated with different concentrations of luteolin and isoorientin.The survival rate of cells was determined by CCK-8 method,and the inhibition rate of α-glucosidase was determined by microplate reader.The mRNA and protein expressions of SGLT-1 and GLUT-2 in Caco-2 cells were detected by RT-PCR and Western blot.Results:The inhibitory effect of 1 μmol/L Luteolin and isoorientin on α-glucosidase was not statistically significant with that of acarbose(P>0.05);50 and 100 μmol/L Luteolin and 100 μmol/L isoorientin SGLT1,GLUT2 mRNA and protein expressions(P<0.05 or P<0.01).Conclusion:Luteolin and isoorientin can delay the hydrolysis of carbohydrates and inhibit the glucose absorption by inhibiting the α-glucosidase enzyme and the mRNA and protein expressions of SGLT1 and GLUT2 in Caco-2 cells.They both have potential hypoglycemic effect.At the same concentration,luteolin is superior to isoorientin.
引文
[1] Jacobs JA,Fetzer SJ.Oral diabetes medications:an update for home health clinicians[J].Home Healthc Nurse,2013,31(1):E1-E13.
[2] Ghani U.Re-exploring promising α-glucosidase inhibitors for potential development into oral anti-diabetic drugs:Finding needle in the haystack[J].European Journal of Medicinal Chemistry,2015,103:133-162.
[3] 张磊,曾丹,郑媛嘉,等.黄芪提取物对结肠腺癌Caco-2细胞葡萄糖吸收的影响[J].中国实验方剂学杂志,2016,22(24):145-149.
[4] Jayaraman R,Subramani S,Sheik Abdullah SH,et al.Antihyperglycemic effect of hesperetin,a citrus flavonoid,extenuates hyperglycemia and exploring the potential role in antioxidant and antihyperlipidemic in streptozotocin-induced diabetic rats[J].Biomedicine & Pharmacotherapy,2018,97:98-106.
[5] Cazarolli LH,Folador P,Moresco HH,et al.Mechanism of action of the stimulatory effect of apigenin-6-C-(2″-O-α-l-rhamnopyranosyl)-β-l-fucopyranoside on 14C-glucose uptake[J].Chemico-Biological Interactions,2009,179(2-3):407-412.
[6] Zhang X,Liu Z,Bi X,et al.Flavonoids and its derivatives from Callistephus chinensis flowers and their inhibitory activities against alpha-glucosidase[J].EXCLI J,2013,12:956-966.
[7] Wu P,He P,Zhao S,et al.Effects of Ursolic Acid Derivatives on Caco-2 Cells and Their Alleviating Role in Streptozocin-Induced Type 2 Diabetic Rats[J].Molecules,2014,19(8):12559-12576.
[8] Satoh T,Igarashi M,Yamada S,et al.Inhibitory effect of black tea and its combination with acarbose on small intestinal α-glucosidase activity[J].Journal of Ethnopharmacology,2015,161:147-155.
[9] Ding H,Hu X,Xu X,et al.Inhibitory mechanism of two allosteric inhibitors,oleanolic acid and ursolic acid on α-glucosidase[J].International Journal of Biological Macromolecules,2018,107:1844-1855.
[10] Yu S,Kwon Y,Lee C,et al.Antidiabetic effect of chitosan oligosaccharide(GO2KA1)is mediated via inhibition of intestinal alpha-glucosidase and glucose transporters and PPARγ expression[J].Bio Factors,2017,43(1):90-99.
[2] Ghani U.Re-exploring promising α-glucosidase inhibitors for potential development into oral anti-diabetic drugs:Finding needle in the haystack[J].European Journal of Medicinal Chemistry,2015,103:133-162.
[3] 张磊,曾丹,郑媛嘉,等.黄芪提取物对结肠腺癌Caco-2细胞葡萄糖吸收的影响[J].中国实验方剂学杂志,2016,22(24):145-149.
[4] Jayaraman R,Subramani S,Sheik Abdullah SH,et al.Antihyperglycemic effect of hesperetin,a citrus flavonoid,extenuates hyperglycemia and exploring the potential role in antioxidant and antihyperlipidemic in streptozotocin-induced diabetic rats[J].Biomedicine & Pharmacotherapy,2018,97:98-106.
[5] Cazarolli LH,Folador P,Moresco HH,et al.Mechanism of action of the stimulatory effect of apigenin-6-C-(2″-O-α-l-rhamnopyranosyl)-β-l-fucopyranoside on 14C-glucose uptake[J].Chemico-Biological Interactions,2009,179(2-3):407-412.
[6] Zhang X,Liu Z,Bi X,et al.Flavonoids and its derivatives from Callistephus chinensis flowers and their inhibitory activities against alpha-glucosidase[J].EXCLI J,2013,12:956-966.
[7] Wu P,He P,Zhao S,et al.Effects of Ursolic Acid Derivatives on Caco-2 Cells and Their Alleviating Role in Streptozocin-Induced Type 2 Diabetic Rats[J].Molecules,2014,19(8):12559-12576.
[8] Satoh T,Igarashi M,Yamada S,et al.Inhibitory effect of black tea and its combination with acarbose on small intestinal α-glucosidase activity[J].Journal of Ethnopharmacology,2015,161:147-155.
[9] Ding H,Hu X,Xu X,et al.Inhibitory mechanism of two allosteric inhibitors,oleanolic acid and ursolic acid on α-glucosidase[J].International Journal of Biological Macromolecules,2018,107:1844-1855.
[10] Yu S,Kwon Y,Lee C,et al.Antidiabetic effect of chitosan oligosaccharide(GO2KA1)is mediated via inhibition of intestinal alpha-glucosidase and glucose transporters and PPARγ expression[J].Bio Factors,2017,43(1):90-99.