蒿甲醚对C57BL/Ks J-db/db小鼠糖脂代谢的影响
|
摘要
目的研究蒿甲醚对db/db小鼠糖脂代谢的影响。方法取8周龄雄性C57BL/KsJ-db/db小鼠,分为模型组(ig给予1%甲基纤维素)和蒿甲醚400、200、100、50组(分别ig给予400、200、100、50 mg/kg蒿甲醚+1%甲基纤维素),每组6只。另选6只雄性C57BL/KsJ-db/+小鼠为对照组,共给药4周。每2天测量小鼠体质量;每3天检测小鼠摄食量并评估平均每日摄食量及体质量变化;每2天测定小鼠饮水量;每3天测量小鼠尿量;每7天于禁食8 h后,尾静脉采血,用罗氏血糖仪及配套试纸分别测量小鼠空腹血糖。通过葡萄糖耐量试验(IPGTT)和胰岛素耐量试验(IPITT)评估小鼠对葡萄糖的耐受力及对胰岛素的敏感性,通过生化试剂盒测定小鼠血清总胆固醇(TC)、三酰甘油(TG)、游离脂肪酸(FFA)水平。HE染色法观察小鼠胰腺和肝脏的形态改变。Westernblotting法分析小鼠肝脏中AMP活化蛋白激酶(AMPK)、葡萄糖转运蛋白体4(GLUT-4)和胰岛素受体β(IRβ)蛋白的表达。结果与对照组比较,模型组小鼠的摄食量、摄水量、尿量均明显升高(P<0.001)。与模型组比较,各剂量蒿甲醚均能显著降低小鼠摄水量及尿量(P<0.01、0.001);蒿甲醚400、200、100 mg/kg均能显著降低小鼠体质量及摄食量,并呈剂量依赖性(P<0.05、0.01);蒿甲醚400、200、100mg/kg均显著降低小鼠空腹血糖水平,减少IPGTT的曲线下面积(AUCs),改善小鼠的胰岛素抵抗(P<0.01、0.001)。与对照组比较,模型组小鼠的TC、TG及FFA水平均显著升高(P<0.05)。与模型组比较,蒿甲醚可显著降低小鼠血清中的TC、TG及FFA水平,并呈剂量依赖性(P<0.05);可显著改善db/db小鼠的胰岛空泡变性和肝脂肪变性,使小鼠肝脏中的AMPK、GLUT-4及IRβ蛋白表达增加(P<0.05)。随着干预时间的延长,剂量越高的小鼠呈现出较高的死亡率及不良反应发生率。结论蒿甲醚可显著改善糖尿病小鼠的高脂状态及胰岛素抵抗,治疗脂肪肝,可能通过AMPK途径上调GLUT-4以及IRβ蛋白的表达发挥作用,有望用于代谢综合征为主的2型糖尿病的治疗。但较高剂量蒿甲醚及较长时间应用会导致较多不良反应的发生。
Objective To investigate the effects of different doses of artemether on glycolipid metabolism in C57 BL/Ks J-db/db mice.Methods Eight-week-old male C57 BL/KsJ-db/db mice were divided into model group(ig given 1% methylcellulose) and artemether 400,200,100,50 groups(ig given 400,200,100,50 mg/kg artemether respectively + 1% methylcellulose),with six mice in each group.Another six male C57 BL/KsJ-db/+ mice were selected as the control group.All groups were administered for 4 weeks.The quality of the mice was measured every 2 d;The food intake of the mice was measured every 3 d and the average daily food intake and body mass changes were evaluated;The amount of water in mice was measured every 2 d;The urine volume of the mice was measured every 3 d;After 8 h of fasting,blood was collected from the tail vein,and the fasting blood glucose of the mice was measured by Roche blood glucose meter and matching test paper every 7 d.Mice were assessed for glucose tolerance and sensitivity to insulin by ip glucose tolerance test(IPGTT) and ip insulin tolerance test(IPITT).The serum total cholesterol(TC),triglyceride(TG),and free fatty acid(FFA) levels of the mice were determined by a biochemical kit.The whole liver of mice was weighed and the morphological changes of the pancreas and liver in mice were observed by HE staining.The expression of AMP-activated protein kinase(AMPK),glucose transporter 4(GLUT-4),and insulin receptor β(IRβ) protein in liver of mice was analyzed by Western blotting.Results Compared with the control group,the food intake,water intake,and urine volume of the model group were significantly increased(P<0.001).Compared with the model group,each dose of artemether significantly reduced the water intake and urine volume of the mice(P<0.01,0.001);Artemether 400,200,100 mg/kg can significantly reduce the body weight and food intake in a dose-dependent manner(P<0.05,0.01);Artemether 400,200,100 mg/kg significantly reduced fasting blood glucose levels in mice,reduced the area under the curve of IPGTT(AUCs),and improved insulin resistance in mice(P<0.01,0.001).Compared with the control group,the TC,TG,and FFA levels of mice in the model group were significantly increased(P<0.05).Compared with the model group,artemether significantly decreased the levels of TC,TG,and FFA in the serum of mice in a dose-dependent manner(P<0.05),and significantly improved islet vacuolar degeneration and hepatic steatosis in db/db mice;The protein expression of AMPK,GLUT-4,and IRβ in the liver of mice was increased(P<0.05).With the prolongation of the intervention time,the higher the dose of artemether,the higher the mortality rate and the incidence of adverse reactions in mice.Conclusion Artemether can significantly improve the high-fat state and insulin resistance of diabetic mice.It can treat fatty liver and may up-regulate the expression of GLUT-4 and IRβ protein through the AMPK pathway to exert its effects.It is expected to be used in the treatment of type 2 diabetes mellitus,which is mainly caused by metabolic syndrome.However,higher doses of artemether and longer-term application may lead to more adverse events.
Objective To investigate the effects of different doses of artemether on glycolipid metabolism in C57 BL/Ks J-db/db mice.Methods Eight-week-old male C57 BL/KsJ-db/db mice were divided into model group(ig given 1% methylcellulose) and artemether 400,200,100,50 groups(ig given 400,200,100,50 mg/kg artemether respectively + 1% methylcellulose),with six mice in each group.Another six male C57 BL/KsJ-db/+ mice were selected as the control group.All groups were administered for 4 weeks.The quality of the mice was measured every 2 d;The food intake of the mice was measured every 3 d and the average daily food intake and body mass changes were evaluated;The amount of water in mice was measured every 2 d;The urine volume of the mice was measured every 3 d;After 8 h of fasting,blood was collected from the tail vein,and the fasting blood glucose of the mice was measured by Roche blood glucose meter and matching test paper every 7 d.Mice were assessed for glucose tolerance and sensitivity to insulin by ip glucose tolerance test(IPGTT) and ip insulin tolerance test(IPITT).The serum total cholesterol(TC),triglyceride(TG),and free fatty acid(FFA) levels of the mice were determined by a biochemical kit.The whole liver of mice was weighed and the morphological changes of the pancreas and liver in mice were observed by HE staining.The expression of AMP-activated protein kinase(AMPK),glucose transporter 4(GLUT-4),and insulin receptor β(IRβ) protein in liver of mice was analyzed by Western blotting.Results Compared with the control group,the food intake,water intake,and urine volume of the model group were significantly increased(P<0.001).Compared with the model group,each dose of artemether significantly reduced the water intake and urine volume of the mice(P<0.01,0.001);Artemether 400,200,100 mg/kg can significantly reduce the body weight and food intake in a dose-dependent manner(P<0.05,0.01);Artemether 400,200,100 mg/kg significantly reduced fasting blood glucose levels in mice,reduced the area under the curve of IPGTT(AUCs),and improved insulin resistance in mice(P<0.01,0.001).Compared with the control group,the TC,TG,and FFA levels of mice in the model group were significantly increased(P<0.05).Compared with the model group,artemether significantly decreased the levels of TC,TG,and FFA in the serum of mice in a dose-dependent manner(P<0.05),and significantly improved islet vacuolar degeneration and hepatic steatosis in db/db mice;The protein expression of AMPK,GLUT-4,and IRβ in the liver of mice was increased(P<0.05).With the prolongation of the intervention time,the higher the dose of artemether,the higher the mortality rate and the incidence of adverse reactions in mice.Conclusion Artemether can significantly improve the high-fat state and insulin resistance of diabetic mice.It can treat fatty liver and may up-regulate the expression of GLUT-4 and IRβ protein through the AMPK pathway to exert its effects.It is expected to be used in the treatment of type 2 diabetes mellitus,which is mainly caused by metabolic syndrome.However,higher doses of artemether and longer-term application may lead to more adverse events.
引文
[1]Nolan C J,Ruderman N B,Kahn S E,et al.Insulin resistance as a physiological defense against metabolic stress:Implications for the management of subsets of type2 diabetes[J].Diabetes,2015,64(3):673-686.
[2]Tahrani A A,Barnett A H,Bailey C J.Pharmacology and therapeutic implications of current drugs for type 2diabetes mellitus[J].Nat Rev Endocrinol,2016,12(10):566-592.
[3]Prabhakar P K,Doble M.A target based therapeutic approach towards diabetes mellitus using medicinal plants[J].Curr Diabetes Rev,2008,4(4):291-308.
[4]Nosten F,White N J.Artemisinin-based combination treatment of falciparum malaria[J].Am J Trop Med Hyg,2007,77(Suppl 6):181-192.
[5]Efferth T.From ancient herb to versatile,modern drug:Artemisia annua and artemisinin for cancer therapy[J].Semin Cancer Biol,2017,46:65-83.
[6]Lu P,Zhang F C,Qian S W,et al.Artemisinin derivatives prevent obesity by inducing browning of WAT and enhancing BAT function[J].Cell Res,2016,26(10):1169-1172.
[7]Lam N S,Long X,Su X Z,et al.Artemisinin and its derivatives in treating helminthic infections beyond schistosomiasis[J].Pharmacol Res,2018,71(7):35-38.
[8]Shi C,Li H,Yang Y,et al.Anti-inflammatory and immunoregulatory functions of artemisinin and its derivatives[J].Mediat Inflamm,2015,doi:10.1155/2015/435713.
[9]Ahmad W,Khan I,Khan M A,et al.Evaluation of antidiabetic and antihyperlipidemic activity of Artemisia indica linn(aeriel parts)in Streptozotocin induced diabetic rats[J].J Ethnopharmacol,2014,151(1):618-623.
[10]Ghazanfar K,Ganai B A,Akbar S,et al.Antidiabetic activity of artemisia amygdalina decne in Streptozotocin induced diabetic rats[J].Biomed Res Int,2015,doi:10.1155/2014/185676.
[11]Li J,Casteels T,Frogne T,et al.Artemisinins target GABAA receptor signaling and impairαcell identity[J].Cell,2017,168(1/2):86-100.
[12]Klayman D L.Qinghasu(artemisinin):An antimalarial drug from China[J].Science,1985,228(4703):1049-1055.
[13]An J,Minie M,Sasaki T,et al.Antimalarial drugs as immune modulators:New mechanisms for old drugs[J].Annual Rev Med,2017,68(1):317-330.
[14]Eun K K,Keon-Hee K,Won H R,et al.Artemisia annua leaf extract attenuates hepatic steatosis and inflammation in high-fat diet-fed mice[J].J Med Food,2016,19(3):290-299.
[15]Guo Y,Fu W,Xin Y,et al.Anti-diabetic and anti-obesity effects of artemether in db/db mice[J].Biomed Res Inter,2018,doi:10.1155/2018/8639523.
[16]Efferth T.Beyond malaria:The inhibition of viruses by artemisinin-type compounds[J].Biotechnol Adv,2018,doi:10.1016/j.biotechadv.2018.01.001.
[17]Jang B C.Artesunate inhibits adipogeneis in 3T3-L1preadipocytes by reducing the expression and/or phosphorylation levels of C/EBP-α,PPAR-γ,FAS,perilipin A,and STAT-3[J].Biochem Biophy Res Commun,2016,474(1):220-225.
[18]Lee J,Kim M H,Lee J H,et al.Artemisinic acid is a regulator of adipocyte differentiation and C/EBPδexpression[J].J Cell Biochem,2012,113(7):2488-2499.
[19]Hardie D G.The AMP-activated protein kinase pathway-new players upstream and downstream[J].JCell Sci,2004,117(23):5479-5487.
[20]Hardie D G.Minireview:The AMP-activated protein kinase cascade:The key sensor of cellular energy status[J].Endocrinology,2003,144(12):5179-5183.
[21]Wang Z Q,Zhang X H,Yu Y,et al.Artemisia scoparia extract attenuates non-alcoholic fatty liver disease in diet-induced obesity mice by enhancing hepatic insulin and AMPK signaling independently of FGF21 pathway[J].Metabol Clin Exper,2013,62(9):1239-1249.
[22]Wu X,An P,Ye B,et al.Artemisinin ameliorated proteinuria in rats with adriamycin-induced nephropathy through regulating nephrin and podocin expressions[J].JTradit Chin Med,2014,34(1):63-68.
[23]Yuan D S,Chen Y P,Tan L L,et al.Artemisinin:Apanacea eligible for unrestrictive use?[J].Front Pharmacol,2017,doi:10.3389/fphar.2017.00737.
[24]Buffet P A,Safeukui I,Deplaine G,et al.The pathogenesis of Plasmodium falciparum malaria in humans:Insights from splenic physiology[J].Blood,2011,117(2):381-392.
[25]White N J.Malaria parasite clearance[J].Malar J,2017,doi:10.1186/s12936-017-1731-1.
[2]Tahrani A A,Barnett A H,Bailey C J.Pharmacology and therapeutic implications of current drugs for type 2diabetes mellitus[J].Nat Rev Endocrinol,2016,12(10):566-592.
[3]Prabhakar P K,Doble M.A target based therapeutic approach towards diabetes mellitus using medicinal plants[J].Curr Diabetes Rev,2008,4(4):291-308.
[4]Nosten F,White N J.Artemisinin-based combination treatment of falciparum malaria[J].Am J Trop Med Hyg,2007,77(Suppl 6):181-192.
[5]Efferth T.From ancient herb to versatile,modern drug:Artemisia annua and artemisinin for cancer therapy[J].Semin Cancer Biol,2017,46:65-83.
[6]Lu P,Zhang F C,Qian S W,et al.Artemisinin derivatives prevent obesity by inducing browning of WAT and enhancing BAT function[J].Cell Res,2016,26(10):1169-1172.
[7]Lam N S,Long X,Su X Z,et al.Artemisinin and its derivatives in treating helminthic infections beyond schistosomiasis[J].Pharmacol Res,2018,71(7):35-38.
[8]Shi C,Li H,Yang Y,et al.Anti-inflammatory and immunoregulatory functions of artemisinin and its derivatives[J].Mediat Inflamm,2015,doi:10.1155/2015/435713.
[9]Ahmad W,Khan I,Khan M A,et al.Evaluation of antidiabetic and antihyperlipidemic activity of Artemisia indica linn(aeriel parts)in Streptozotocin induced diabetic rats[J].J Ethnopharmacol,2014,151(1):618-623.
[10]Ghazanfar K,Ganai B A,Akbar S,et al.Antidiabetic activity of artemisia amygdalina decne in Streptozotocin induced diabetic rats[J].Biomed Res Int,2015,doi:10.1155/2014/185676.
[11]Li J,Casteels T,Frogne T,et al.Artemisinins target GABAA receptor signaling and impairαcell identity[J].Cell,2017,168(1/2):86-100.
[12]Klayman D L.Qinghasu(artemisinin):An antimalarial drug from China[J].Science,1985,228(4703):1049-1055.
[13]An J,Minie M,Sasaki T,et al.Antimalarial drugs as immune modulators:New mechanisms for old drugs[J].Annual Rev Med,2017,68(1):317-330.
[14]Eun K K,Keon-Hee K,Won H R,et al.Artemisia annua leaf extract attenuates hepatic steatosis and inflammation in high-fat diet-fed mice[J].J Med Food,2016,19(3):290-299.
[15]Guo Y,Fu W,Xin Y,et al.Anti-diabetic and anti-obesity effects of artemether in db/db mice[J].Biomed Res Inter,2018,doi:10.1155/2018/8639523.
[16]Efferth T.Beyond malaria:The inhibition of viruses by artemisinin-type compounds[J].Biotechnol Adv,2018,doi:10.1016/j.biotechadv.2018.01.001.
[17]Jang B C.Artesunate inhibits adipogeneis in 3T3-L1preadipocytes by reducing the expression and/or phosphorylation levels of C/EBP-α,PPAR-γ,FAS,perilipin A,and STAT-3[J].Biochem Biophy Res Commun,2016,474(1):220-225.
[18]Lee J,Kim M H,Lee J H,et al.Artemisinic acid is a regulator of adipocyte differentiation and C/EBPδexpression[J].J Cell Biochem,2012,113(7):2488-2499.
[19]Hardie D G.The AMP-activated protein kinase pathway-new players upstream and downstream[J].JCell Sci,2004,117(23):5479-5487.
[20]Hardie D G.Minireview:The AMP-activated protein kinase cascade:The key sensor of cellular energy status[J].Endocrinology,2003,144(12):5179-5183.
[21]Wang Z Q,Zhang X H,Yu Y,et al.Artemisia scoparia extract attenuates non-alcoholic fatty liver disease in diet-induced obesity mice by enhancing hepatic insulin and AMPK signaling independently of FGF21 pathway[J].Metabol Clin Exper,2013,62(9):1239-1249.
[22]Wu X,An P,Ye B,et al.Artemisinin ameliorated proteinuria in rats with adriamycin-induced nephropathy through regulating nephrin and podocin expressions[J].JTradit Chin Med,2014,34(1):63-68.
[23]Yuan D S,Chen Y P,Tan L L,et al.Artemisinin:Apanacea eligible for unrestrictive use?[J].Front Pharmacol,2017,doi:10.3389/fphar.2017.00737.
[24]Buffet P A,Safeukui I,Deplaine G,et al.The pathogenesis of Plasmodium falciparum malaria in humans:Insights from splenic physiology[J].Blood,2011,117(2):381-392.
[25]White N J.Malaria parasite clearance[J].Malar J,2017,doi:10.1186/s12936-017-1731-1.