芒果苷对高糖诱导β细胞凋亡的影响及机制研究
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摘要
目的:研究芒果苷对胰岛β细胞的保护作用及并探讨其可能的作用机制。方法:建立高糖诱导INS-1细胞凋亡模型,用不同浓度的芒果苷预处理细胞,MTT法检测细胞凋亡,Western blot检测凋亡相关蛋白Bax、Bcl-2表达,荧光探针法检测细胞内ROS水平,丙二醛(MDA)检测试剂盒检测细胞内MDA水平。结果:33 mmol/L葡萄糖能显著诱导INS-1细胞凋亡,芒果苷能以浓度依赖性保护INS-1细胞,降低细胞凋亡率,同时能恢复高糖引起的Bcl-2蛋白表达降低,降低Bax蛋白表达;33 mmol/L葡萄糖能诱导INS-1细胞产生ROS及MDA,芒果苷处理细胞能显著降低ROS及MDA水平。结论:芒果苷可能通过降低细胞内氧化压力抑制高糖诱导的INS-1细胞凋亡。
Objective:To study the protective effect and possible mechanism of mangiferin on pancreatic β cells induced by high glucose.Methods:Apoptosis model of INS-1 cells induced by high glucose were established.The cells were pretreated with mangiferin at different concentrations.The apoptosis was determined by MTT assay,and the apoptosis-related protein expressions of Bax and Bcl-2 were detected by Western blot.The level of ROS was determined by fluorescent probe,and MDA was determined by MDA detection kit.Results:The apoptosis of INS-1 cells was induced by 33 mmol/L glucose,mangiferin could protect INS-1 cells in a concentration-dependent manner and reduce apoptosis rate and the expression of Bax,improve the expression of Bcl-2 at the same time.33 mmol/L glucose could induce INS-1 cells to produce ROS and MDA,while mangiferin-treated cells significantly reduced ROS and MDA levels.Conclusion:Mangiferin may inhibit hyperglycemic-indnced apoptosis of INs-1 cells by reducing intracellular oxidative stress.
Objective:To study the protective effect and possible mechanism of mangiferin on pancreatic β cells induced by high glucose.Methods:Apoptosis model of INS-1 cells induced by high glucose were established.The cells were pretreated with mangiferin at different concentrations.The apoptosis was determined by MTT assay,and the apoptosis-related protein expressions of Bax and Bcl-2 were detected by Western blot.The level of ROS was determined by fluorescent probe,and MDA was determined by MDA detection kit.Results:The apoptosis of INS-1 cells was induced by 33 mmol/L glucose,mangiferin could protect INS-1 cells in a concentration-dependent manner and reduce apoptosis rate and the expression of Bax,improve the expression of Bcl-2 at the same time.33 mmol/L glucose could induce INS-1 cells to produce ROS and MDA,while mangiferin-treated cells significantly reduced ROS and MDA levels.Conclusion:Mangiferin may inhibit hyperglycemic-indnced apoptosis of INs-1 cells by reducing intracellular oxidative stress.
引文
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[10]吴巍,滕厚雷.小檗碱与芒果苷及其配伍对L6成纤维细胞葡萄糖摄取的影响及机制[J].中国药理学通报,2010,26(9):1259-1260.
[11] Arozal W,Suyatna FD,Juniantito V,et al.The Effects of Mangiferin(Mangifera indica L)in Doxorubicin-induced Cardiotoxicity in Rats[J]. Drug Res(Stuttg),2015,65(11):574-580.
[12]殷万斐,彭志刚,赵卫华,等.芒果苷对柔红霉素所致大鼠心肌细胞损伤的影响及机制研究[J].中药材,2013,36(4):626-629.
[13] Luo Y,Fu C,Wang Z,et al.Mangiferin attenuates contusive spinal cord injury in rats through the regulation of oxidative stress,inflammation and the Bcl-2 and Bax pathway[J].Mol Med Rep,2015,12(5):7132-7138.
[14] He L,Peng X,Zhu J,et al.Mangiferin attenuate sepsis-induced acute kidney injury via antioxidant and anti-inflammatory effects[J].Am J Nephrol,2014,40(5):441-450.
[15] Yang Z,Weian C,Susu H,et al.Protective effects of mangiferin on cerebral ischemia-reperfusion injury and its mechanisms[J].Eur J Pharmacol,2016,771:145-151.
[2] Robertson RP.β-Cell deterioration during diabetes:what's in the gun?[J]. Trends Endocrinol Metab,2009,20(8):388-393.
[3] Inoguchi T,Li P,Umeda F,et al. High glucose level and free fatty acid stimulate reactive oxygen species production through protein kinase C-dependent activation of NAD(P)H oxidase in cultured vascular cells[J]. Diabetes,2000,49(11):1939-1945.
[4] Hamaoka R,Fujii J,Miyagawa J,et al.Overexpression of the aldose reductase gene induces apoptosis in pancreaticβ-cells by causing a redox imbalance[J]. J Biochem,1999,126(1):41-47.
[5] Kaneto H,Xu G,Song KH,et al. Activation of the hexosamine pathway leads to deterioration of pancreaticβ-cell function through the induction of oxidative stress[J].J Biol Chem,2001,276(33):31099-31104.
[6] Nishikawa T,Edelstein D,Du XL,et al.Normalizing mitochondrial super-oxide production blocks three pathways of hyperglycaemic damage[J]. Nature,2000,404(6779):787-790.
[7] Sakai K,Matsumoto K,Nishikawa T,et al.Mitochondrial reactive oxygen species reduce insulin secretion by pancreaticβ-cells[J]. Biochem Biophys Res Commun,2003,300(1):216-222.
[8]郭宏伟,邓家刚,运晨霞,等.芒果苷抑制哮喘小鼠气道炎症的机制[J].中国实验方剂学杂志,2012,18(9):187-190.
[9]谭之强.芒果苷通过抑制氧化应激及抗凋亡作用保护大鼠颅脑损伤[J].神经损伤与功能重建,2018,13(1):26-28.
[10]吴巍,滕厚雷.小檗碱与芒果苷及其配伍对L6成纤维细胞葡萄糖摄取的影响及机制[J].中国药理学通报,2010,26(9):1259-1260.
[11] Arozal W,Suyatna FD,Juniantito V,et al.The Effects of Mangiferin(Mangifera indica L)in Doxorubicin-induced Cardiotoxicity in Rats[J]. Drug Res(Stuttg),2015,65(11):574-580.
[12]殷万斐,彭志刚,赵卫华,等.芒果苷对柔红霉素所致大鼠心肌细胞损伤的影响及机制研究[J].中药材,2013,36(4):626-629.
[13] Luo Y,Fu C,Wang Z,et al.Mangiferin attenuates contusive spinal cord injury in rats through the regulation of oxidative stress,inflammation and the Bcl-2 and Bax pathway[J].Mol Med Rep,2015,12(5):7132-7138.
[14] He L,Peng X,Zhu J,et al.Mangiferin attenuate sepsis-induced acute kidney injury via antioxidant and anti-inflammatory effects[J].Am J Nephrol,2014,40(5):441-450.
[15] Yang Z,Weian C,Susu H,et al.Protective effects of mangiferin on cerebral ischemia-reperfusion injury and its mechanisms[J].Eur J Pharmacol,2016,771:145-151.