D-松醇复配Mn~(2+)对2型糖尿病大鼠的降血糖作用及其机制的研究
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摘要
目的:研究D-松醇复配Mn~(2+)对2型糖尿病大鼠的降血糖作用并探讨其作用机制。方法:选用SPF级雄性大鼠进行实验,高糖高脂饲料喂养配合注射链脲佐菌素(STZ)诱导建立2型糖尿病(T2DM)模型。实验动物分为空白对照组、模型对照组、阳性对照组、D-松醇组和低、中、高复配Mn~(2+)组,灌胃期间每周定时测体重和空腹血糖(FBG),4周后测定口服葡萄糖耐量(OGTT)和胰岛素耐量(ITT),5周后处死并对其糖化血清蛋白(GSP)、空腹血清胰岛素(FINS)等生理生化指标进行测定,计算胰岛素敏感性指数(ISI)和胰岛β细胞功能指数(HOMA-β)。并采用实时荧光全定量分析(RT-PCR)实验检测AMPK信号通路相关基因mRNA表达水平。结果:与模型对照组相比,剂量组均可显著性降低T2DM大鼠空腹血糖值(p<0.05),高剂量复配组血糖值极显著降低(p<0.01)。此外,与D-松醇组相比,高剂量复配组的FINS和GSP极显著降低(p<0.01)。与模型对照组相比,ISI-1、ISI-2和AMPKα-2基因mRNA表达水平显著上调(p<0.05),且高剂量组与D-松醇组相比极显著性上调(p<0.01)。高剂量复配组AMPKα-1和GLUT4基因mRNA表达水平显著上调(p<0.05),PGC-1α、PEPCK和G6Pase基因mRNA表达水平显著下调(p<0.05)。结论:D-松醇复配Mn~(2+)提高了T2DM大鼠对葡萄糖的耐受能力,缓解胰岛素抵抗症状,提高机体对胰岛素的敏感性,其作用机制可能涉及AMPK参与的抑制糖异生等生化调控过程起到降血糖作用。
Objective:The objective of this work was to explore the hypoglycemic effects of D-pinitol compound with Mn~(2+) on type 2 diabetes mellitus rats and underling mechanism. Methods:A model of type 2 diabetes mellitus (T2 DM) was established by feeding high-fat diet and streptozocin (STZ) injecting to male rats. The rats were divided into the normal group,control group,positive control group,D-pinitol group,low,medium and high complex Mn~(2+) group.Body weight and fasting blood glucose (FBG) were tested each week. Oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) was tested after four-week-treatment. The contents of glycated serum protein (GSP) and fasting insulin (FINS) were determined after five-week-treatment and insulin sensitivity index (ISI) and pancreatic β cell function index (HOMA-β) were calculated. The expression of AMPK genes was detected with real-time polymerase chain reaction (real-time PCR). Results:Compared with the model group,the hypoglycemic effects of dosage groups increased significantly(p<0.05),and the high combined group increased significantly (p<0.01).Compared with the D-pinitol group,the FINS and GSP of the high combined group increased significantly. Compared with the model group,the mRNA expression levels of ISI-1,ISI-2 and AMPKα-2 increased significantly(p<0.05),and the high combined group increased significantly (p<0.01).The mRNA expression levels of AMPKα-1 and GLUT4 increased significantly (p<0.05)while PGC-1α,PEPCK and G6pase mRNA expression decreased significantly (p<0.05)in the high combined group. Conclusion:D-pinitol compound with Mn~(2+) could improve the body's sensitivity to insulin,moreover,it has a positive effect on hypoglycemic activity through AMPK signaling pathway.
Objective:The objective of this work was to explore the hypoglycemic effects of D-pinitol compound with Mn~(2+) on type 2 diabetes mellitus rats and underling mechanism. Methods:A model of type 2 diabetes mellitus (T2 DM) was established by feeding high-fat diet and streptozocin (STZ) injecting to male rats. The rats were divided into the normal group,control group,positive control group,D-pinitol group,low,medium and high complex Mn~(2+) group.Body weight and fasting blood glucose (FBG) were tested each week. Oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) was tested after four-week-treatment. The contents of glycated serum protein (GSP) and fasting insulin (FINS) were determined after five-week-treatment and insulin sensitivity index (ISI) and pancreatic β cell function index (HOMA-β) were calculated. The expression of AMPK genes was detected with real-time polymerase chain reaction (real-time PCR). Results:Compared with the model group,the hypoglycemic effects of dosage groups increased significantly(p<0.05),and the high combined group increased significantly (p<0.01).Compared with the D-pinitol group,the FINS and GSP of the high combined group increased significantly. Compared with the model group,the mRNA expression levels of ISI-1,ISI-2 and AMPKα-2 increased significantly(p<0.05),and the high combined group increased significantly (p<0.01).The mRNA expression levels of AMPKα-1 and GLUT4 increased significantly (p<0.05)while PGC-1α,PEPCK and G6pase mRNA expression decreased significantly (p<0.05)in the high combined group. Conclusion:D-pinitol compound with Mn~(2+) could improve the body's sensitivity to insulin,moreover,it has a positive effect on hypoglycemic activity through AMPK signaling pathway.
引文
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[2]陈松景.2型糖尿病高危人群干预指标与效果分析方法研究[D].北京:北京理工大学,2014.
[3]黄建华.2型糖尿病相关的代谢组学和生物信息学研究[D].长沙:中南大学,2014.
[4]蔡玲.夜关门中 D-松醇的提取分离及纯化[D].长沙:中南大学,2011:31-38.
[5]Cháfer A,Berna A.Study of kinetics of the D-pinitol extraction from carob pods using supercritical CO2[J].Journal of Supercritical Fluids,2014,94:212-215.
[6]Poongothai G,Sripathi SK.A review on insulin mimetic pinitol from plants[J].International Journal of Pharma and Bio Sciences,2013,4:992-1009.
[7]Livesey G.Health potential of polyols as sugar replacers,with emphasis on low glycaemic properties[J].Nutrition Research Reviews,2003,16(2):163-191.
[8]Ananthan B,Chang WC,Lin JS,et al.A C2Symmetric Chiral Pool-Based Flexible Strategy:Synthesis of(+)-and(-)-Shikimic Acids;(+)-and(-)-4-epi-Shikimic Acids,and(+)-and(-)-Pinitol[J].The Journal of Organic Chemistry,2014,79(7):2898-2905.
[9]Hudlicky T,Price JD,Fan R,et al.Efficient and enantiodivergent synthesis of(+)-and(-)-pinitol[J].Journal of American Chemical Society,1990,112(25):9439-9440.
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[11]Kim M J,Yoo K H,Kim J H,et al.Effect of pinitol on glucose metabolism and adipocytokines in uncontrolled type 2 diabetes[J].Diabetes Research & Clinical Practice,2007,77(3):S247-51.
[12]Bates SH.Insulin-like effect of pinitol[J].British Journal of Pharmacology,2000,130(8):1944-1948.
[13]Sivakumar S,Subramanian SP.Pancreatic tissue protective nature of D-Pinitol studied in streptozotocin-mediated oxidative stress in experimental diabetic rats[J].European Journal of Pharmacology,2009,622(1-3):65-70.
[14]Sivakumar S,Palsamy P,Subramania SP,et al.Impact of D-pinitol on the attenuation of prion flammatory cytokines,hyperglycemia-mediated oxidative stress and protection of kidney tissue ultrastructure in streptozotocin-induced diabetic rats[J].Chemico-Biological Interactions,2010,188(1):237-245.
[15]Larner J.D-Chiro-Inositol glycans in insulin signaling and insulin resistance[J].Molecular Medicine,2010,16(11-12):543-551.
[16]高云峰.D-手型肌醇和D-松醇的降血糖机理研究[D].天津:天津科技大学,2016.
[17]高山.沙棘果油中棕榈油酸的提取及其功能特性评价[D].天津:天津科技大学,2017.
[18]黎宇,罗欣欣,严奉东,等.葛根上调肝胰岛素抵抗HepG2细胞OB-R,IRS2,GLUT1和GLUT2蛋白调节糖代谢的研究[J],中国中药杂志,2017,42(10):1939-1944.
[19]罗招凡,李芳萍,程桦.抵抗素对HepG2肝细胞胰岛素信号通路IRS-2/Akt的影响[J].中国老年学杂志,2016,36(15):3629-3631.
[20]Hyun Jin Kim,Kang Seo Park,Seong Kyu Lee,et al.Effects of pinitol on glycemic control,insulin resistance and adipocytokine levels in patients with type 2 diabetes mellitus[J].Nutrition & Metabolism,2012,60(3):1-5.
[21]Rengarajan T,Rajendran P,Nandakumar N,et al.Free radical scavenging and antioxidant activity of D-pinitol against 7,12 dimethylbenz(a)anthracene induced breast cancer in sprague dawley rats[J].Asian Pacific Journal Tropical Disease,2014,4(5):384-390.
[22]尹忞强,刘溪,吴婷婷,等.羁糖脂对HepG2胰岛素抵抗细胞葡萄糖代谢及相关蛋白表达的影响[J].药物评价研究,2017,40(6):783-787.
[23]汪莎.野木瓜皂苷抗糖尿病活性及其作用机制[D].武汉:中南民族大学,2015.
[24]Long YC,Zierath JR.AMP-activated protein kinase signaling in metabolic regulation[J].J Clin Invest,2006,116(7):1776-1783.
[25]Rengarajan T,Nandakumar N,Periyasamy MB.D-Pinitol prevents rat breast carcinogenesis induced by 7,12-Dimethylbenz[a]anthracene through inhibition of Bcl-2 and induction of p53,caspase-3 proteins and modulation of hepatic biotransformation enzymes and antioxidants[J].Biomedicine & Preventive Nutrition,2013,3(1):31-41.
[26]Amit D Kandhare1,Subhash L Bodhankar1,Vishwaraman Mohan,et al.Prophylactic efficacy and possible mechanisms of oligosaccharides based standardized fenugreek seed extract on high-fat diet-induced insulin resistance in C57BL/6 mice[J].Pediatr Gastroenterol Nutr,2015,5(3):35-45.
[2]陈松景.2型糖尿病高危人群干预指标与效果分析方法研究[D].北京:北京理工大学,2014.
[3]黄建华.2型糖尿病相关的代谢组学和生物信息学研究[D].长沙:中南大学,2014.
[4]蔡玲.夜关门中 D-松醇的提取分离及纯化[D].长沙:中南大学,2011:31-38.
[5]Cháfer A,Berna A.Study of kinetics of the D-pinitol extraction from carob pods using supercritical CO2[J].Journal of Supercritical Fluids,2014,94:212-215.
[6]Poongothai G,Sripathi SK.A review on insulin mimetic pinitol from plants[J].International Journal of Pharma and Bio Sciences,2013,4:992-1009.
[7]Livesey G.Health potential of polyols as sugar replacers,with emphasis on low glycaemic properties[J].Nutrition Research Reviews,2003,16(2):163-191.
[8]Ananthan B,Chang WC,Lin JS,et al.A C2Symmetric Chiral Pool-Based Flexible Strategy:Synthesis of(+)-and(-)-Shikimic Acids;(+)-and(-)-4-epi-Shikimic Acids,and(+)-and(-)-Pinitol[J].The Journal of Organic Chemistry,2014,79(7):2898-2905.
[9]Hudlicky T,Price JD,Fan R,et al.Efficient and enantiodivergent synthesis of(+)-and(-)-pinitol[J].Journal of American Chemical Society,1990,112(25):9439-9440.
[10]詹天荣,娄红祥.右旋肌醇甲醚的植物资源与医药功效[J].国外医药植物药分册,2006,21(1):17-19.
[11]Kim M J,Yoo K H,Kim J H,et al.Effect of pinitol on glucose metabolism and adipocytokines in uncontrolled type 2 diabetes[J].Diabetes Research & Clinical Practice,2007,77(3):S247-51.
[12]Bates SH.Insulin-like effect of pinitol[J].British Journal of Pharmacology,2000,130(8):1944-1948.
[13]Sivakumar S,Subramanian SP.Pancreatic tissue protective nature of D-Pinitol studied in streptozotocin-mediated oxidative stress in experimental diabetic rats[J].European Journal of Pharmacology,2009,622(1-3):65-70.
[14]Sivakumar S,Palsamy P,Subramania SP,et al.Impact of D-pinitol on the attenuation of prion flammatory cytokines,hyperglycemia-mediated oxidative stress and protection of kidney tissue ultrastructure in streptozotocin-induced diabetic rats[J].Chemico-Biological Interactions,2010,188(1):237-245.
[15]Larner J.D-Chiro-Inositol glycans in insulin signaling and insulin resistance[J].Molecular Medicine,2010,16(11-12):543-551.
[16]高云峰.D-手型肌醇和D-松醇的降血糖机理研究[D].天津:天津科技大学,2016.
[17]高山.沙棘果油中棕榈油酸的提取及其功能特性评价[D].天津:天津科技大学,2017.
[18]黎宇,罗欣欣,严奉东,等.葛根上调肝胰岛素抵抗HepG2细胞OB-R,IRS2,GLUT1和GLUT2蛋白调节糖代谢的研究[J],中国中药杂志,2017,42(10):1939-1944.
[19]罗招凡,李芳萍,程桦.抵抗素对HepG2肝细胞胰岛素信号通路IRS-2/Akt的影响[J].中国老年学杂志,2016,36(15):3629-3631.
[20]Hyun Jin Kim,Kang Seo Park,Seong Kyu Lee,et al.Effects of pinitol on glycemic control,insulin resistance and adipocytokine levels in patients with type 2 diabetes mellitus[J].Nutrition & Metabolism,2012,60(3):1-5.
[21]Rengarajan T,Rajendran P,Nandakumar N,et al.Free radical scavenging and antioxidant activity of D-pinitol against 7,12 dimethylbenz(a)anthracene induced breast cancer in sprague dawley rats[J].Asian Pacific Journal Tropical Disease,2014,4(5):384-390.
[22]尹忞强,刘溪,吴婷婷,等.羁糖脂对HepG2胰岛素抵抗细胞葡萄糖代谢及相关蛋白表达的影响[J].药物评价研究,2017,40(6):783-787.
[23]汪莎.野木瓜皂苷抗糖尿病活性及其作用机制[D].武汉:中南民族大学,2015.
[24]Long YC,Zierath JR.AMP-activated protein kinase signaling in metabolic regulation[J].J Clin Invest,2006,116(7):1776-1783.
[25]Rengarajan T,Nandakumar N,Periyasamy MB.D-Pinitol prevents rat breast carcinogenesis induced by 7,12-Dimethylbenz[a]anthracene through inhibition of Bcl-2 and induction of p53,caspase-3 proteins and modulation of hepatic biotransformation enzymes and antioxidants[J].Biomedicine & Preventive Nutrition,2013,3(1):31-41.
[26]Amit D Kandhare1,Subhash L Bodhankar1,Vishwaraman Mohan,et al.Prophylactic efficacy and possible mechanisms of oligosaccharides based standardized fenugreek seed extract on high-fat diet-induced insulin resistance in C57BL/6 mice[J].Pediatr Gastroenterol Nutr,2015,5(3):35-45.