桑叶黄酮对糖尿病大鼠血糖水平的影响及机制探讨
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摘要
目的:通过观察桑叶黄酮降血糖作用,探讨其作用机制。方法:8周龄雄性SD大鼠24只,链脲佐菌素(STZ)腹腔注射造模成功后,按血糖、体质量随机分为模型组、吡格列酮组、桑叶黄酮组,每组8只,另设同周龄正常SD大鼠8只为正常组。治疗6周后,检测大鼠的空腹血糖(fasting blood glucose,FBG),总胆固醇(cholesterd,CHO),甘油三酯(total triglyceride,TG),游离脂肪酸(free fatty acids,FFA),天门冬氨酸氨基转移酶(aspartate aminotransferase,AST),血清胰岛素水平(fasting insulin,Fins),计算胰岛素抵抗指数(homa insulin-resistance,HOMA-IR);采用实时荧光定量PCR,蛋白免疫印迹法(Western blot)检测肝脏相关mRNA和蛋白表达。结果:与正常组比较,模型组大鼠FBG,Fins,HOMA-IR,体质量,CHO,TG,FFA明显升高(P<0.05,P<0.01);肝脏蛋白激酶C(protein kinase C,PKC)mRNA和蛋白表达明显升高(P<0.05,P<0.01),腺苷酸激酶2(adenylate kinase2,AK2),过氧化物酶增殖物激活受体γ辅助激活因子1α(peroxisome proliferator activated receptor gamma coactivator 1-alpha,PGC1α)mRNA表达明显降低(P<0.05,P<0.01)。与模型组比较,桑叶黄酮组FBG,Fins,HOMA-IR,体质量,CHO,TG,FFA明显降低(P<0.05,P<0.01);肝脏PKC mRNA和蛋白表达明显降低(P<0.05,P<0.01),AK2,PGC-1αmRNA表达明显升高(P<0.05,P<0.01)。结论:桑叶黄酮具有降低血糖,FFA的作用,机制可能是通过抑制FFA诱导的PKC途径以及改善基于AK2,PGC1α表达的能量稳态发挥抗糖尿病效果。
Objective: To elucidate the effect and mechanism of flavonoids from Mori Folium(FM) on improving blood glucose in diabetic rats. Method: After successful modeling by intraperitoneal injection of streptozotocin(STZ),twenty-four 8-week-old male SD rats were randomly divided into model group,Pioglitazone group,FM group with 8 rats in each group according to their levels of blood glucose and body weight,and another8 normal SD rats with the same age were also used as a normal group. After treatment for 6 weeks,their fasting blood glucose(FBG), cholesterd(CHO), total triglyceride(TG), free fatty acids(FFA), aspartate aminotransferase(AST),fasting insulin(Fins),and homa insulin-resistance(HOMA-IR) were measured. Realtime fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot were used to quantify the mRNA and protein expressions of certain targets in the liver. Result: As compared with the normal group,the body weight,FBG,Fins,HOMA-IR,CHO,TG,FFA were significantly increased in model group(P < 0. 05,P < 0. 01); the mRNA and protein expressions of protein kinase C(PKC) were increased significantly in model group(P < 0. 05, P < 0. 01), but the mRNA expressions of adenylate kinase2(AK2), and peroxisome proliferator activated receptor gamma coactivator 1-alpha(PGC-1α) were decreased significantly(P < 0. 05,P <0. 01). As compared with the model group,body weight,FBG,Fins,HOMA-IR,CHO,TG,and FFA were significantly decreased in FM group(P < 0. 05,P < 0. 01); the mRNA and protein expressions of PKC in liver tissues were decreased significantly(P < 0. 05,P < 0. 01),but the mRNA expressions of AK2,and PGC-1α were increased significantly in FM group(P < 0. 05,P < 0. 01). Conclusion: It was demonstrated that FM may play an anti-diabetic effect by inhibiting the PKC pathway induced by FFA and improving the energy homeostasis based on the expression of AK2 and PGC-1α.
Objective: To elucidate the effect and mechanism of flavonoids from Mori Folium(FM) on improving blood glucose in diabetic rats. Method: After successful modeling by intraperitoneal injection of streptozotocin(STZ),twenty-four 8-week-old male SD rats were randomly divided into model group,Pioglitazone group,FM group with 8 rats in each group according to their levels of blood glucose and body weight,and another8 normal SD rats with the same age were also used as a normal group. After treatment for 6 weeks,their fasting blood glucose(FBG), cholesterd(CHO), total triglyceride(TG), free fatty acids(FFA), aspartate aminotransferase(AST),fasting insulin(Fins),and homa insulin-resistance(HOMA-IR) were measured. Realtime fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot were used to quantify the mRNA and protein expressions of certain targets in the liver. Result: As compared with the normal group,the body weight,FBG,Fins,HOMA-IR,CHO,TG,FFA were significantly increased in model group(P < 0. 05,P < 0. 01); the mRNA and protein expressions of protein kinase C(PKC) were increased significantly in model group(P < 0. 05, P < 0. 01), but the mRNA expressions of adenylate kinase2(AK2), and peroxisome proliferator activated receptor gamma coactivator 1-alpha(PGC-1α) were decreased significantly(P < 0. 05,P <0. 01). As compared with the model group,body weight,FBG,Fins,HOMA-IR,CHO,TG,and FFA were significantly decreased in FM group(P < 0. 05,P < 0. 01); the mRNA and protein expressions of PKC in liver tissues were decreased significantly(P < 0. 05,P < 0. 01),but the mRNA expressions of AK2,and PGC-1α were increased significantly in FM group(P < 0. 05,P < 0. 01). Conclusion: It was demonstrated that FM may play an anti-diabetic effect by inhibiting the PKC pathway induced by FFA and improving the energy homeostasis based on the expression of AK2 and PGC-1α.
引文
[1]Federation I D.International diabetes federation diabetes atlas seventh edition 2015[M].International Diabetes Federation:Seventh Edition,2016:9-12.
[2]黄链莎,刘铜华,孙文,等.桂皮醛对糖尿病小鼠血糖水平的影响及机制[J].中国实验方剂学杂志,2018,24(10):95-100.
[3]ZHENG Y,Ley S H,HU F B.Global aetiology and epidemiology of type 2 diabetes mellitus and its complications[J].Nat Rev Endocrinol,2017,doi:10.1038/nrendo.
[4]田思敏,柳辰玥,马双双,等.桑叶对糖尿病小鼠肝脏Toll样受体基因表达的影响[J].中国实验方剂学杂志,2017,23(6):137-142.
[5]Riche D M,Riche K D,East H E,et al.Impact of mulberry leaf extract on type 2 diabetes(Mul-DM):a randomized,placebo-controlled pilot study[J].Complement Ther Med,2017,32:105-108.
[6]HONG C T,LIU Z H,HUANG Y T,et al.Extract from Mulberry(Morus australis)leaf decelerate acetaminophen induced hepatic inflammation involving downregulation of myeloid differentiation factor 88(My D88)signals[J].J Food Drug Anal,2017,25(4):862-871.
[7]季涛,宿树兰,郭盛,等.桑叶防治糖尿病的效应成分群及其作用机制研究进展[J].中草药,2015,46(5):778-784.
[8]孟庆海,殷秋忆,郭静,等.4种不同桑叶提取物降血糖作用的筛选[J].中成药,2014,36(6):1288-1291.
[9]王文文,张赛,朱晓卉,等.桑叶总黄酮对1型糖尿病小鼠肾间质纤维化的防治作用及机制[J].中国药理学通报,2017,33(9):1278-1285.
[10]张立雯,季涛,宿树兰,等.桑叶黄酮类和生物碱类成分在正常和糖尿病大鼠体内的药代动力学研究[J].中国中药杂志,2017,42(21):4218-4225.
[11]CAI S,SUN W,FAN Y,et al.Effect of mulberry leaf(Folium Mori)on insulin resistance via IRS-1/PI3K/Glut-4 signalling pathway in type 2 diabetes mellitus rats[J].Pharm Biol,2016,54(11):2685-2691.
[12]范愈燕,吕翠岩,朱斌,等.黄芪甲苷对链脲佐菌素诱导糖尿病大鼠肾脏保护机制探讨[J].中华中医药杂志,2017,32(1):294-298.
[13]葛争艳,金龙,杨斌,等.降糖消脂片对KK-A~y转基因小鼠胰岛素抵抗的影响[J].中国中西医结合杂志,2017,37(3):331-337.
[14]SHENG Y,ZHENG S,MA T,et al.Mulberry leaf alleviates streptozotocin-induced diabetic rats by attenuating NEFA signaling and modulating intestinal microflora[J].Sci Rep,2017,7(1):12041.
[15]Tillander V,Alexson S E H,Cohen D E.Deactivating fatty acids:acyl-Co A thioesterase-mediated control of lipid metabolism[J].Trends Endocrinol Metab,2017,28(7):473-484.
[16]Bachnio-Zabielska A,Zabielski P,Baranowski M,et al.Effects of streptozotocin-induced diabetes and elevation of plasma FFA on ceramide metabolism in rat skeletal muscle[J].Horm Metab Res,2010,42(1):1-7.
[17]Santomauro A T,Boden G,Silva M E,et al.Overnight lowering of free fatty acids with Acipimox improves insulin resistance and glucose tolerance in obese diabetic and nondiabetic subjects[J].Diabetes,1999,48(9):1836-1841.
[18]Sharma D,Garg S,Mehndiratta M,et al.Relationship of serum apolipoprotein A-V levels,oxidative stress and inflammatory biomarkers with hypertriglyceridemia in type 2 diabetes mellitus[J].Int J Endocrinol Metab,2017,15(2):e44805.
[19]孙文佳,杜方岭,刘玮,等.对高糖损伤HBZY-1细胞具有保护作用的天然植物提取物的筛选及其抗炎活性研究[J].现代食品科技,2017,33(5):33-38,76.
[20]黄立佳,杜月光,孙梽均,等.桑叶黄酮和多糖对糖尿病大鼠血清肿瘤坏死因子-α、C反应蛋白含量及肾功能的影响[J].中国老年学杂志,2016,36(19):4703-4705.
[21]XIONG M,HUANG Y,LIU Y,et al.Antidiabetic activity of Eergosterol from pleurotus ostreatus in KK-Ay mice with spontaneous type 2 diabetes mellitus[J].Mol Nutr Food Res,2017,doi:10.1002/mnfr.201700444.
[22]Sajan M P,Hansen B C,Higgs M G,et al.Atypical PKC,PKCλ/ι,activatesβ-secretase and increases Aβ1-40/42 and phospho-tau in mouse brain and isolated neuronal cells,and may link hyperinsulinemia and other a PKC activators to development of pathological and memory abnormalities in Alzheimer's disease[J].Neurobiol Aging,2018,61:225-237.
[23]Prentki M,Joly E,El-Assaad W,et al.Malonyl-Co A signaling,lipid partitioning,and glucolipotoxicity:role in beta-cell adaptation and failure in the etiology of diabetes[J].Diabetes,2002,51(Suppl 3):S405-S413.
[24]Pereira S,Park E,Mori Y,et al.FFA-induced hepatic insulin resistance in vivo is mediated by PKCδ,NADPH oxidase,and oxidative stress[J].Am J Physiol Endocrinol Metab,2014,307(1):E34-E46.
[25]Crunkhorn S.Metabolic disease:PGC1αinhibition ameliorates diabetes[J].Nat Rev Drug Discov,2017,16(6):386.
[26]Moreno-Santos I,Pérez-Belmonte L M,MacíasGonzález M,et al.Type 2 diabetes is associated with decreased PGC1αexpression in epicardial adipose tissue of patients with coronary artery disease[J].J Transl Med,2016,14(1):243.
[2]黄链莎,刘铜华,孙文,等.桂皮醛对糖尿病小鼠血糖水平的影响及机制[J].中国实验方剂学杂志,2018,24(10):95-100.
[3]ZHENG Y,Ley S H,HU F B.Global aetiology and epidemiology of type 2 diabetes mellitus and its complications[J].Nat Rev Endocrinol,2017,doi:10.1038/nrendo.
[4]田思敏,柳辰玥,马双双,等.桑叶对糖尿病小鼠肝脏Toll样受体基因表达的影响[J].中国实验方剂学杂志,2017,23(6):137-142.
[5]Riche D M,Riche K D,East H E,et al.Impact of mulberry leaf extract on type 2 diabetes(Mul-DM):a randomized,placebo-controlled pilot study[J].Complement Ther Med,2017,32:105-108.
[6]HONG C T,LIU Z H,HUANG Y T,et al.Extract from Mulberry(Morus australis)leaf decelerate acetaminophen induced hepatic inflammation involving downregulation of myeloid differentiation factor 88(My D88)signals[J].J Food Drug Anal,2017,25(4):862-871.
[7]季涛,宿树兰,郭盛,等.桑叶防治糖尿病的效应成分群及其作用机制研究进展[J].中草药,2015,46(5):778-784.
[8]孟庆海,殷秋忆,郭静,等.4种不同桑叶提取物降血糖作用的筛选[J].中成药,2014,36(6):1288-1291.
[9]王文文,张赛,朱晓卉,等.桑叶总黄酮对1型糖尿病小鼠肾间质纤维化的防治作用及机制[J].中国药理学通报,2017,33(9):1278-1285.
[10]张立雯,季涛,宿树兰,等.桑叶黄酮类和生物碱类成分在正常和糖尿病大鼠体内的药代动力学研究[J].中国中药杂志,2017,42(21):4218-4225.
[11]CAI S,SUN W,FAN Y,et al.Effect of mulberry leaf(Folium Mori)on insulin resistance via IRS-1/PI3K/Glut-4 signalling pathway in type 2 diabetes mellitus rats[J].Pharm Biol,2016,54(11):2685-2691.
[12]范愈燕,吕翠岩,朱斌,等.黄芪甲苷对链脲佐菌素诱导糖尿病大鼠肾脏保护机制探讨[J].中华中医药杂志,2017,32(1):294-298.
[13]葛争艳,金龙,杨斌,等.降糖消脂片对KK-A~y转基因小鼠胰岛素抵抗的影响[J].中国中西医结合杂志,2017,37(3):331-337.
[14]SHENG Y,ZHENG S,MA T,et al.Mulberry leaf alleviates streptozotocin-induced diabetic rats by attenuating NEFA signaling and modulating intestinal microflora[J].Sci Rep,2017,7(1):12041.
[15]Tillander V,Alexson S E H,Cohen D E.Deactivating fatty acids:acyl-Co A thioesterase-mediated control of lipid metabolism[J].Trends Endocrinol Metab,2017,28(7):473-484.
[16]Bachnio-Zabielska A,Zabielski P,Baranowski M,et al.Effects of streptozotocin-induced diabetes and elevation of plasma FFA on ceramide metabolism in rat skeletal muscle[J].Horm Metab Res,2010,42(1):1-7.
[17]Santomauro A T,Boden G,Silva M E,et al.Overnight lowering of free fatty acids with Acipimox improves insulin resistance and glucose tolerance in obese diabetic and nondiabetic subjects[J].Diabetes,1999,48(9):1836-1841.
[18]Sharma D,Garg S,Mehndiratta M,et al.Relationship of serum apolipoprotein A-V levels,oxidative stress and inflammatory biomarkers with hypertriglyceridemia in type 2 diabetes mellitus[J].Int J Endocrinol Metab,2017,15(2):e44805.
[19]孙文佳,杜方岭,刘玮,等.对高糖损伤HBZY-1细胞具有保护作用的天然植物提取物的筛选及其抗炎活性研究[J].现代食品科技,2017,33(5):33-38,76.
[20]黄立佳,杜月光,孙梽均,等.桑叶黄酮和多糖对糖尿病大鼠血清肿瘤坏死因子-α、C反应蛋白含量及肾功能的影响[J].中国老年学杂志,2016,36(19):4703-4705.
[21]XIONG M,HUANG Y,LIU Y,et al.Antidiabetic activity of Eergosterol from pleurotus ostreatus in KK-Ay mice with spontaneous type 2 diabetes mellitus[J].Mol Nutr Food Res,2017,doi:10.1002/mnfr.201700444.
[22]Sajan M P,Hansen B C,Higgs M G,et al.Atypical PKC,PKCλ/ι,activatesβ-secretase and increases Aβ1-40/42 and phospho-tau in mouse brain and isolated neuronal cells,and may link hyperinsulinemia and other a PKC activators to development of pathological and memory abnormalities in Alzheimer's disease[J].Neurobiol Aging,2018,61:225-237.
[23]Prentki M,Joly E,El-Assaad W,et al.Malonyl-Co A signaling,lipid partitioning,and glucolipotoxicity:role in beta-cell adaptation and failure in the etiology of diabetes[J].Diabetes,2002,51(Suppl 3):S405-S413.
[24]Pereira S,Park E,Mori Y,et al.FFA-induced hepatic insulin resistance in vivo is mediated by PKCδ,NADPH oxidase,and oxidative stress[J].Am J Physiol Endocrinol Metab,2014,307(1):E34-E46.
[25]Crunkhorn S.Metabolic disease:PGC1αinhibition ameliorates diabetes[J].Nat Rev Drug Discov,2017,16(6):386.
[26]Moreno-Santos I,Pérez-Belmonte L M,MacíasGonzález M,et al.Type 2 diabetes is associated with decreased PGC1αexpression in epicardial adipose tissue of patients with coronary artery disease[J].J Transl Med,2016,14(1):243.