槐属二氢黄酮G促进L6细胞内GLUT4的表达和转运
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摘要
利用细胞葡萄糖摄取检测试剂盒检测苦参中槐属二氢黄酮G(Sophoraflavanone G,SFG)对L6细胞葡萄糖摄取的影响,发现SFG能够增加大鼠成肌细胞(L6)的葡萄糖摄取;之后,使用Western blot检测发现SFG对L6细胞内GLUT4的表达有显著的促进作用;同时,在可稳定表达IRAP-mOrange荧光蛋白的L6细胞内,使用激光共聚焦显微镜监测SFG作用下葡萄糖转位蛋白4 (glucose transporter 4,GLUT4)的转位,发现SFG对GLUT4的转位有显著的促进作用,而且SFG对GLUT4转位的促进呈浓度依赖性;另外,免疫荧光实验结果也显示SFG增强L6细胞中GLUT4与细胞膜的融合.这些结果显示利用SFG处理L6细胞后,L6细胞内GLUT4的表达、转运及与细胞膜的融合继而促进葡萄糖的摄取显著增强,说明SFG可能具备开发成一种新的降血糖药物的潜力.
In this study,the effects of Sophoraflavanone G(SFG) on glucose uptake were detected in L6 cells by cell glucose uptake assay kit. We found that SFG increased glucose uptake in L6 cells. Western blotting was used to detect the effects of SFG on GLUT4 expression level, and it was found that SFG significantly promoted GLUT4 expression in L6 cells. Meanwhile, the translocation of GLUT4 also was monitored. We found that SFG significantly promoted GLUT4 translocation, and the effects of SFG on the translocation of GLUT4 was concentration dependent in L6 cells. In addition, immunofluorescence test results also showed that SFG enhanced the fusion of GLUT4 and cell membrane in L6 cells.These results showed that SFG promoted glucose uptake by improving GLUT4 expression, translocation and fusion with cell membrane in L6 cell and SFG might have the potential to develop a new hypoglycemic drug.
In this study,the effects of Sophoraflavanone G(SFG) on glucose uptake were detected in L6 cells by cell glucose uptake assay kit. We found that SFG increased glucose uptake in L6 cells. Western blotting was used to detect the effects of SFG on GLUT4 expression level, and it was found that SFG significantly promoted GLUT4 expression in L6 cells. Meanwhile, the translocation of GLUT4 also was monitored. We found that SFG significantly promoted GLUT4 translocation, and the effects of SFG on the translocation of GLUT4 was concentration dependent in L6 cells. In addition, immunofluorescence test results also showed that SFG enhanced the fusion of GLUT4 and cell membrane in L6 cells.These results showed that SFG promoted glucose uptake by improving GLUT4 expression, translocation and fusion with cell membrane in L6 cell and SFG might have the potential to develop a new hypoglycemic drug.
引文
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[15] JIANG L,FAN J,BAI L,et al.Direct quantification of fusion rate reveals a distal role for AS160 in insulin-stimulated fusion of GLUT4 storage vesicles[J].J Biol Chem,2008,283(13):8508-8516.
[16] LI Y,ZHENG L,WANG D,et al.Staurosporine as an agonist for induction of GLUT4 translocation,identified by a pH-sensitive fluorescent IRAP-mOrange2 probe[J].Biochem Biophys Res Commun,2016,480(4):534-538.
[17] ALBISTON A L,CACADOR M,SINNAYAH P,et al.Insulin-regulated aminopeptidase inhibitors do not alter glucose handling in normal and diabetic rats[J].J Mol Endocrinol,2017,58(4):193-198.
[18] FADINI G P,BOTTIGLIENGO D,D’ANGELO F,et al.Comparative effectiveness of DPP-4 inhibitors versus sulfonylurea for the treatment of type 2 diabetes in routine clinical practice:a retrospective multicenter real-world study[J].Diabetes Ther,2018,9(4):1477-1490.
[19] TANABE M,NOMIYAMA T,MOTONAGA R,et al.Reduced vascular events in type 2 diabetes by biguanide relative to sulfonylurea:study in a Japanese Hospital Database[J].BMC Endocr Disord,2015,15:49.
[20] JOSHI S R,STANDL E,TONG N,et al.Therapeutic potential of alpha-glucosidase inhibitors in type 2 diabetes mellitus:an evidence-based review[J].Expert Opin Pharmacother,2015,16(13):1959-1981.
[21] ZHOU J,LI H,ZHANG X,et al.Nateglinide and acarbose are comparably effective reducers of postprandial glycemic excursions in chinese antihyperglycemic agent-naive subjects with type 2 diabetes[J].Diabetes Technol Ther,2013,15(6):481-488.
[22] LI W L,ZHENG H C,BUKURU J,et al.Natural medicines used in the traditional Chinese medical system for therapy of diabetes mellitus[J].J Ethnopharmacol,2004,92(1):1-21.
[2] SHIH C C,LIN C H,LIN W L,et al.Momordica charantia extract on insulin resistance and the skeletal muscle GLUT4 protein in fructose-fed rats[J].J Ethnopharmacol,2009,123(1):82-90.
[3] ZHANG C,JIANG Y,LIU J,et al.AMPK/AS160 mediates tiliroside derivatives-stimulated GLUT4 translocation in muscle cells[J].Drug Des Devel Ther,2018,12:1581-1587.
[4] VLAVCHESKI F,BARON D,VLACHOGIANNIS I A,et al.Carnosol increases skeletal muscle cell glucose uptake via AMPK-dependent GLUT4 glucose transporter translocation[J].Int J Mol Sci,2018,19(5):1321.
[5] TALANTIKITE M,BERENGUER M,GONZALES T,et al.The first intracellular loop of GLUT4 contains a retention motif[J].J Cell Sci,2016,129(11):2273-2284.
[6] CHAKRABORTY C,BANDYOPADHYAY S,MAULIK U,et al.Topology mapping of insulin-regulated glucose transporter GLUT4 using computational biology[J].Cell Biochem Biophys,2013,67(3):1261-1274.
[7] LACOMBE V A.Expression and regulation of facilitative glucose transporters in equine insulin-sensitive tissue:from physiology to pathology[J].ISRN Vet Sci,2014,2014:409547 (1-15).
[8] 张翅,马悦,高慧敏,等.苦参化学成分研究进展[J].中国实验方剂学杂志,2014,20(4):205-214.ZHANG C,MA Y,GAO H M,et al.Advance on chemical constituents in Sophora flavescens[J].Chinese Journal of Experimental Traditional Medical Formulae,2014,20(4):205-214.(Ch).
[9] 杨志欣,单柏松,李霞,等.槐属二氢黄酮G的药理活性及构效关系评述[J].中药材,2016,39(2):457-461.YANG Z X,SHAN B S,LI X,et al.Pharmacological activity and structure-activity relationship of sophoraflavanone G [J].Journal of Chinese Medicinal Materials,2016,39(2):457-461.(Ch).
[10] KIM C S,PARK S N,AHN S J,et al.Antimicrobial effect of sophoraflavanone G isolated from Sophora flavescens against mutans streptococci[J].Anaerobe,2013,19:17-21.
[11] KIM B H,WON C,LEE Y H,et al.Sophoraflavanone G induces apoptosis of human cancer cells by targeting upstream signals of STATs[J].Biochem Pharmacol,2013,86(7):950-959.
[12] GUO C,YANG L,WAN C X,et al.Anti-neuroinflammatory effect of Sophoraflavanone G from Sophora alopecuroides in LPS-activated BV2 microglia by MAPK,JAK/STAT and Nrf2/HO-1 signaling pathways[J].Phytomedicine,2016,23(13):1629-1637.
[13] WANG H,CHEN L,ZHANG L,et al.Protective effect of Sophoraflavanone G on streptozotocin (STZ)-induced inflammation in diabetic rats[J].Biomed Pharmacother,2016,84:1617-1622.
[14] ABEL E D,GRAVELEAU C,BETUING S,et al.Regulation of insulin-responsive aminopeptidase expression and targeting in the insulin-responsive vesicle compartment of glucose transporter isoform 4-deficient cardiomyocytes[J].Mol Endocrinol,2004,18(10):2491-2501.
[15] JIANG L,FAN J,BAI L,et al.Direct quantification of fusion rate reveals a distal role for AS160 in insulin-stimulated fusion of GLUT4 storage vesicles[J].J Biol Chem,2008,283(13):8508-8516.
[16] LI Y,ZHENG L,WANG D,et al.Staurosporine as an agonist for induction of GLUT4 translocation,identified by a pH-sensitive fluorescent IRAP-mOrange2 probe[J].Biochem Biophys Res Commun,2016,480(4):534-538.
[17] ALBISTON A L,CACADOR M,SINNAYAH P,et al.Insulin-regulated aminopeptidase inhibitors do not alter glucose handling in normal and diabetic rats[J].J Mol Endocrinol,2017,58(4):193-198.
[18] FADINI G P,BOTTIGLIENGO D,D’ANGELO F,et al.Comparative effectiveness of DPP-4 inhibitors versus sulfonylurea for the treatment of type 2 diabetes in routine clinical practice:a retrospective multicenter real-world study[J].Diabetes Ther,2018,9(4):1477-1490.
[19] TANABE M,NOMIYAMA T,MOTONAGA R,et al.Reduced vascular events in type 2 diabetes by biguanide relative to sulfonylurea:study in a Japanese Hospital Database[J].BMC Endocr Disord,2015,15:49.
[20] JOSHI S R,STANDL E,TONG N,et al.Therapeutic potential of alpha-glucosidase inhibitors in type 2 diabetes mellitus:an evidence-based review[J].Expert Opin Pharmacother,2015,16(13):1959-1981.
[21] ZHOU J,LI H,ZHANG X,et al.Nateglinide and acarbose are comparably effective reducers of postprandial glycemic excursions in chinese antihyperglycemic agent-naive subjects with type 2 diabetes[J].Diabetes Technol Ther,2013,15(6):481-488.
[22] LI W L,ZHENG H C,BUKURU J,et al.Natural medicines used in the traditional Chinese medical system for therapy of diabetes mellitus[J].J Ethnopharmacol,2004,92(1):1-21.