Spleen-kidney supplementing formula alleviates insulin resistance via regulating AKT/glycogen synthase kinase 3β pathway in rats with type 2 diabetic induced by high-fat diet
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摘要
OBJECTIVE: To explore the molecular mechanism underpinning the action by investigating its effect on glycogen content and AKT(also known as protein kinase B)/glycogen synthase kinase 3β(GSK-3β) pathway in the liver of rats with type 2 diabetic induced by high-fat diet.METHODS: The rat model of type 2 diabetes was induced by high-fat diet and multiple low-dose streptozotocin injection. Diabetic rats were divided into five groups: the model control group, the Metformin group, spleen-kidney supplementing formula groups of low, medium and high doses. Fasting blood glucose(FBG) levels were measured before treatment and every two weeks during treatment.After the treatment, oral glucose tolerance test was performed, and hemoglobin A1c (HbA1c) and C-peptide were measured to assess the formula's effect on glucose metabolism and insulin resistance. The protein expression levels of AKT, GSK-3βand their phosphorylated forms in the liver were also measured to study the formula's role in insulin signaling pathway.RESULTS: Spleen-kidney supplementing formula significantly relieved the symptoms of polydipsia,polyuria and weight loss in type 2 diabetic rats, reduced FBG and HbA1c levels, increased glycogen content, and improved insulin sensitivity. The anti-diabetic effects of spleen-kidney supplementing formula are dose dependent. It also increased the total AKT protein level and the GSK-3β phosphorylation in the liver of type 2 diabetic rats.CONCLUSION: Spleen-kidney supplementing formula has hypoglycemic effect and relieves insulin resistance by enhancing AKT/GSK-3β signaling pathway in the liver of type 2 diabetic rats.
OBJECTIVE: To explore the molecular mechanism underpinning the action by investigating its effect on glycogen content and AKT(also known as protein kinase B)/glycogen synthase kinase 3β(GSK-3β) pathway in the liver of rats with type 2 diabetic induced by high-fat diet.METHODS: The rat model of type 2 diabetes was induced by high-fat diet and multiple low-dose streptozotocin injection. Diabetic rats were divided into five groups: the model control group, the Metformin group, spleen-kidney supplementing formula groups of low, medium and high doses. Fasting blood glucose(FBG) levels were measured before treatment and every two weeks during treatment.After the treatment, oral glucose tolerance test was performed, and hemoglobin A1c (HbA1c) and C-peptide were measured to assess the formula's effect on glucose metabolism and insulin resistance. The protein expression levels of AKT, GSK-3βand their phosphorylated forms in the liver were also measured to study the formula's role in insulin signaling pathway.RESULTS: Spleen-kidney supplementing formula significantly relieved the symptoms of polydipsia,polyuria and weight loss in type 2 diabetic rats, reduced FBG and HbA1c levels, increased glycogen content, and improved insulin sensitivity. The anti-diabetic effects of spleen-kidney supplementing formula are dose dependent. It also increased the total AKT protein level and the GSK-3β phosphorylation in the liver of type 2 diabetic rats.CONCLUSION: Spleen-kidney supplementing formula has hypoglycemic effect and relieves insulin resistance by enhancing AKT/GSK-3β signaling pathway in the liver of type 2 diabetic rats.
OBJECTIVE: To explore the molecular mechanism underpinning the action by investigating its effect on glycogen content and AKT(also known as protein kinase B)/glycogen synthase kinase 3β(GSK-3β) pathway in the liver of rats with type 2 diabetic induced by high-fat diet.METHODS: The rat model of type 2 diabetes was induced by high-fat diet and multiple low-dose streptozotocin injection. Diabetic rats were divided into five groups: the model control group, the Metformin group, spleen-kidney supplementing formula groups of low, medium and high doses. Fasting blood glucose(FBG) levels were measured before treatment and every two weeks during treatment.After the treatment, oral glucose tolerance test was performed, and hemoglobin A1c (HbA1c) and C-peptide were measured to assess the formula's effect on glucose metabolism and insulin resistance. The protein expression levels of AKT, GSK-3βand their phosphorylated forms in the liver were also measured to study the formula's role in insulin signaling pathway.RESULTS: Spleen-kidney supplementing formula significantly relieved the symptoms of polydipsia,polyuria and weight loss in type 2 diabetic rats, reduced FBG and HbA1c levels, increased glycogen content, and improved insulin sensitivity. The anti-diabetic effects of spleen-kidney supplementing formula are dose dependent. It also increased the total AKT protein level and the GSK-3β phosphorylation in the liver of type 2 diabetic rats.CONCLUSION: Spleen-kidney supplementing formula has hypoglycemic effect and relieves insulin resistance by enhancing AKT/GSK-3β signaling pathway in the liver of type 2 diabetic rats.
引文
1 International Diabetes Federation.Diabetes Atlas Eighth Edition,2017-12-01,cited 2018-03-21.Available from URL:http://www.diabetesatlas.org/resources/2017-atlas.html,2017:9.
2 Xu Y,Wang LM,He J,et al.Prevalence and control of diabetes in Chinese adults.JAMA 2013;310(9):948-959.
3 La WY,Bai SF,La XJ.Chinese-English edition a collection of classical formulae for diabetes.Beijing:People's Medical Publishing House,2011:2-189.
4 You H,Zhang T,Feng W,et al.Association of TCM body constitution with insulin resistance and risk of diabetes in impaired glucose regulation patients.BMC Complement Altern Med 2017;17(1):459.
5 Chen L,Yang Z,Chen W,et al.Differential expression of immune-related genes between healthy volunteers and type 2 diabetic patients with spleen-deficiency pattern.JTradit Chin Med 2015;35(6):646-652.
6 Kirkman SM,Briscoe VJ,Clark N,et al.Diabetes in older adults:a consensus report.J Am Geriatr Soc 2012;60(12):2342-2356.
7 Jiang N,Liu HF,Li SD,et al.An integrated metabonomic and proteomic study on Kidney-Yin deficiency syndrome patients with diabetes mellitus in China.Acta Pharmacologica Sinica 2015;36(6):689-698.
8 Lan J,Zhao Y,Dong F,et al.Meta-analysis of the effect and safety of berberine in the treatment of type 2 diabetes mellitus,hyperlipemia and hypertension.J Ethnopharmacol 2015;161:69-81.
9 She S,Liu W,Li T,et al.Effects of puerarin in STZ-induced diabetic rats by oxidative stress and the TGF-β1/Smad2 pathway.Food Funct 2014;5(5):944-950.
10 Thorburn AW,Gumbiner B,Bulacan F,et al.Intracellular glucose oxidation and glycogen synthase activity are reduced in non-insulin-dependent(typeⅡ)diabetes independent of impaired glucose uptake.J Clin Invest 1990;85(2):522-529.
11 Cohen P.The Croonian Lecture 1998.Identification of a protein kinase cascade of major importance in insulin signal transduction.Philos T R Soc A 1999;354(1382):485-495.
12 Woodgett JR.Molecular cloning and expression of glycogen synthase kinase-3/factor A.Embo J 1990;9(8):2431-2438.
13 Lappas M.GSK3βIs increased in adipose tissue and skeletal muscle from women with gestational diabetes where it regulates the inflammatory response.PLoS One 2014;9(12):e115854.
14 Zhang T,Fang M,Fu ZM,et al.Expression of PI3-K,PKB and GSK-3βin the skeletal muscle tissue of gestational diabetes mellitus.Asian Pac J Trop Med 2014;7(4):309-312.
15 Motawi TM,Bustanji Y,El-Maraghy SA,et al.Naproxen and cromolyn as new glycogen synthase kinase 3βinhibitors for amelioration of diabetes and obesity:an investigation by docking simulation and subsequent in vitro/in vivo biochemical evaluation.J Biochem Mol Toxic 2013;27(9):425-436.
16 Welsh GI,Proud CG.Glycogen synthase kinase-3 is rapidly inactivated in response to insulin and phosphorylates eukaryotic initiation factor eIF-2B.Biochem J 1993;294(Pt 3):625-629.
17 Cross DA,Alessi DR,Cohen P,et al.Hemmin inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B.Nature 1995;378(6559):785-789.
18 Tian CY,Bo HM,La XJ,et al.Optimized berberine extraction from spleen-kidney supplementing formula by orthogonal tests.Hebei Lian He Da Xue Xue Bao Yi Xue Ban 2013;15(1):51-52.
19 Qian C,Zhu C,Yu W,et al.High-fat diet/low-dose streptozotocin-induced type 2 diabetes in rats impacts osteogenesis and wnt signaling in bone marrow stromal cells.PLoSOne 2015;10(8):e0136390.
20 Liu DH,Xing XF.Evaluation and characteristic of rat model of type 2 diabetes.Zhong Guo Shi Yan Fang Ji Xue Za Zhi 2010;16(12):212-214.
21 Van Miert ASJPAM,Bogaert MG,Debackere M.Comparative veterinary pharmacology,toxicology and therapy.Lancaster:MTP Press Limited,1986:489-500.
22 Li X,Zhou ZG,Qi HY,et al.Replacement of insulin by fasting C-peptide in modified homeostasis model assessment to evaluate insulin resistance and isletβcell function.Zhong Nan Da Xue Xue Bao Yi Xue Ban 2004;29(4):419-423.
23 Li GW,Fan XR.A new insulin-sensitivity index for the population-based study.Chin J Intern Medvol 1993;32(10):656-660.
24 Saini V.Molecular mechanisms of insulin resistance in type 2 diabetes mellitus.World J Diabetes 2010;1(3):68-75.
25 Weyer C,Tataranni PA,Bogardus C,et al.Insulin resistance and insulin secretory dysfunction are independent predictors of worsening of glucose tolerance during each stage of type 2 diabetes development.Diabetes Care 2001;24(1):89-94.
26 Fonseca VA.Defining and characterizing the progression of type 2 diabetes.Diabetes Care 2009;32(Suppl 2):S151-S156.
27 Jacob PS,de Meneses Fujii TM,Yamada M,et al.Isocaloric intake of a high-fat diet promotes insulin resistance and inflammation in Wistar rats.Cell Biochem Funct 2013;31(3):244-253.
28 Srinivasan K,Viswanad B,Asrat L,et al.Combination of high-fat diet-fed and low-dose streptozotocin-treated rat:a model for type 2 diabetes and pharmacological screening.Pharmacol Res 2005;52(4):313-320.
29 Zhou J,Zhou S,Tang J,et al.Protective effect of berberine on beta cells in streptozotocin-and high-carbohydrate/high-fat diet-induced diabetic rats.Eur J Pharmacol 2009;606(1-3):262-268.
30 Yang L,Yao D,Yang H,et al.Puerarin protects pancreaticβ-cells in obese diabetic mice via activation of glp-1r signaling.Molecular Endocrinology 2016;30(3):361-371.
2 Xu Y,Wang LM,He J,et al.Prevalence and control of diabetes in Chinese adults.JAMA 2013;310(9):948-959.
3 La WY,Bai SF,La XJ.Chinese-English edition a collection of classical formulae for diabetes.Beijing:People's Medical Publishing House,2011:2-189.
4 You H,Zhang T,Feng W,et al.Association of TCM body constitution with insulin resistance and risk of diabetes in impaired glucose regulation patients.BMC Complement Altern Med 2017;17(1):459.
5 Chen L,Yang Z,Chen W,et al.Differential expression of immune-related genes between healthy volunteers and type 2 diabetic patients with spleen-deficiency pattern.JTradit Chin Med 2015;35(6):646-652.
6 Kirkman SM,Briscoe VJ,Clark N,et al.Diabetes in older adults:a consensus report.J Am Geriatr Soc 2012;60(12):2342-2356.
7 Jiang N,Liu HF,Li SD,et al.An integrated metabonomic and proteomic study on Kidney-Yin deficiency syndrome patients with diabetes mellitus in China.Acta Pharmacologica Sinica 2015;36(6):689-698.
8 Lan J,Zhao Y,Dong F,et al.Meta-analysis of the effect and safety of berberine in the treatment of type 2 diabetes mellitus,hyperlipemia and hypertension.J Ethnopharmacol 2015;161:69-81.
9 She S,Liu W,Li T,et al.Effects of puerarin in STZ-induced diabetic rats by oxidative stress and the TGF-β1/Smad2 pathway.Food Funct 2014;5(5):944-950.
10 Thorburn AW,Gumbiner B,Bulacan F,et al.Intracellular glucose oxidation and glycogen synthase activity are reduced in non-insulin-dependent(typeⅡ)diabetes independent of impaired glucose uptake.J Clin Invest 1990;85(2):522-529.
11 Cohen P.The Croonian Lecture 1998.Identification of a protein kinase cascade of major importance in insulin signal transduction.Philos T R Soc A 1999;354(1382):485-495.
12 Woodgett JR.Molecular cloning and expression of glycogen synthase kinase-3/factor A.Embo J 1990;9(8):2431-2438.
13 Lappas M.GSK3βIs increased in adipose tissue and skeletal muscle from women with gestational diabetes where it regulates the inflammatory response.PLoS One 2014;9(12):e115854.
14 Zhang T,Fang M,Fu ZM,et al.Expression of PI3-K,PKB and GSK-3βin the skeletal muscle tissue of gestational diabetes mellitus.Asian Pac J Trop Med 2014;7(4):309-312.
15 Motawi TM,Bustanji Y,El-Maraghy SA,et al.Naproxen and cromolyn as new glycogen synthase kinase 3βinhibitors for amelioration of diabetes and obesity:an investigation by docking simulation and subsequent in vitro/in vivo biochemical evaluation.J Biochem Mol Toxic 2013;27(9):425-436.
16 Welsh GI,Proud CG.Glycogen synthase kinase-3 is rapidly inactivated in response to insulin and phosphorylates eukaryotic initiation factor eIF-2B.Biochem J 1993;294(Pt 3):625-629.
17 Cross DA,Alessi DR,Cohen P,et al.Hemmin inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B.Nature 1995;378(6559):785-789.
18 Tian CY,Bo HM,La XJ,et al.Optimized berberine extraction from spleen-kidney supplementing formula by orthogonal tests.Hebei Lian He Da Xue Xue Bao Yi Xue Ban 2013;15(1):51-52.
19 Qian C,Zhu C,Yu W,et al.High-fat diet/low-dose streptozotocin-induced type 2 diabetes in rats impacts osteogenesis and wnt signaling in bone marrow stromal cells.PLoSOne 2015;10(8):e0136390.
20 Liu DH,Xing XF.Evaluation and characteristic of rat model of type 2 diabetes.Zhong Guo Shi Yan Fang Ji Xue Za Zhi 2010;16(12):212-214.
21 Van Miert ASJPAM,Bogaert MG,Debackere M.Comparative veterinary pharmacology,toxicology and therapy.Lancaster:MTP Press Limited,1986:489-500.
22 Li X,Zhou ZG,Qi HY,et al.Replacement of insulin by fasting C-peptide in modified homeostasis model assessment to evaluate insulin resistance and isletβcell function.Zhong Nan Da Xue Xue Bao Yi Xue Ban 2004;29(4):419-423.
23 Li GW,Fan XR.A new insulin-sensitivity index for the population-based study.Chin J Intern Medvol 1993;32(10):656-660.
24 Saini V.Molecular mechanisms of insulin resistance in type 2 diabetes mellitus.World J Diabetes 2010;1(3):68-75.
25 Weyer C,Tataranni PA,Bogardus C,et al.Insulin resistance and insulin secretory dysfunction are independent predictors of worsening of glucose tolerance during each stage of type 2 diabetes development.Diabetes Care 2001;24(1):89-94.
26 Fonseca VA.Defining and characterizing the progression of type 2 diabetes.Diabetes Care 2009;32(Suppl 2):S151-S156.
27 Jacob PS,de Meneses Fujii TM,Yamada M,et al.Isocaloric intake of a high-fat diet promotes insulin resistance and inflammation in Wistar rats.Cell Biochem Funct 2013;31(3):244-253.
28 Srinivasan K,Viswanad B,Asrat L,et al.Combination of high-fat diet-fed and low-dose streptozotocin-treated rat:a model for type 2 diabetes and pharmacological screening.Pharmacol Res 2005;52(4):313-320.
29 Zhou J,Zhou S,Tang J,et al.Protective effect of berberine on beta cells in streptozotocin-and high-carbohydrate/high-fat diet-induced diabetic rats.Eur J Pharmacol 2009;606(1-3):262-268.
30 Yang L,Yao D,Yang H,et al.Puerarin protects pancreaticβ-cells in obese diabetic mice via activation of glp-1r signaling.Molecular Endocrinology 2016;30(3):361-371.