Effects of Eucommia ulmoides Leaves on Glucose Metabolism and Related Gene Expression in Sheep Liver
GAO Yan1, YANG Gaiqing2, WANG Linfeng1, DU Qingxin3, WANG Wenhui4, FU Tong1, LIAN Hongxia1, GAO Tengyun1, LI Ming1
1. College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China;
2. Advanced Experimental Techniques and Management Centre, Henan Agricultural University, Zhengzhou 450002, China;
3. Economic Forestry Research and Development Center, Chinese Academy of Forestry Sciences Zhengzhou 450003, China;
4. Henan Genetic Performance Measurement Center for Breeding Cattle, Zhengzhou 450002, China
Abstract：In order to research the effects of Eucommia ulmoides leaves (EUL) on glucose metabolism and related genes expression in sheep live, 12 healthy sheep (dorper sheep♂×Hu lambs♀) with the similar age (6 to 8-month-old) and body weight (between 35 to 40 kg) were selected and divided into two groups randomly and equally (6 sheep in each group), which named control group (CTL group) and EUL group (diet contained 10% of EUL). The adaption period was 7 days, and the experiment period was 30 days. At the end of experiment, blood samples were collected to determine related biochemical indexes and hormones. After the experiment, all sheep were slaughtered and liver tissues were collected to transcriptome sequence, determine genes and protein expression levels for enzymes and factors involved in glucose metabolism. The results showed as follows:1) compared with CTL group, plasma glucose and serum glucagon contents in EUL group significantly decreased (P<0.05), whereas serum insulin content increased significantly (P<0.05). 2) The results of transcriptome analysis showed that the differentially significant genes related to glucose metabolism and energy metabolism, and significantly enriched in the glucose metabolic pathways. 3) Fluorescent quantitative PCR results showed that the mRNA expression levels of insulin receptor (INSR) and insulin receptor substrate 2 (IRS2) increased significantly (P<0.01), the mRNA expression level of glucagon receptor (GCGR) dropped markedly (P<0.01); the phosphofructokinase (PFKL), the key enzyme in the glycolytic pathway, mRNA expression level up-regulated notably (P<0.05); on the gluconeogenesis pathway, the mRNA expression level of glucose-6-phosphatase (G6Pase) decreased significantly (P<0.01) and the mRNA expression level of phosphoenolpyruvate carboxykinase 1 (PEPCK1) decreased significantly (P<0.05); the mRNA expression levels of fork head transcription factor 1 (FoxO1) and cyclic adenosine monophosphate response element binding protein (CREB) decreased significantly (P<0.01); furthermore, glycogen synthase 2 (Gys2), which promoted glycogen synthesis, mRNA expression increased significantly (P<0.01). 4) Western blot results showed that compared with CTL group, the protein expression level of INSR in EUL group had no significant change (P>0.05), but the protein expression levels of FoxO1、G6Pase and PEPCK1 decreased significantly (P<0.01). In conclusion, EUL greatly affects glucose metabolism of sheep, increases the expression of the key enzyme genes for glycolysis, down-regulates the expression of gluconeogenesis related nuclear transcription factors and enzymes, promotes the synthesis of liver glycogen, and then decreases blood glucose.
高燕, 杨改青, 王林枫, 杜庆鑫, 王文辉, 付彤, 廉红霞, 高腾云, 李明. 杜仲叶对绵羊肝脏糖代谢及其相关基因表达的影响[J]. 动物营养学报, 2019, 31(6): 2854-2864.
GAO Yan, YANG Gaiqing, WANG Linfeng, DU Qingxin, WANG Wenhui, FU Tong, LIAN Hongxia, GAO Tengyun, LI Ming. Effects of Eucommia ulmoides Leaves on Glucose Metabolism and Related Gene Expression in Sheep Liver. Chinese Journal of Animal Nutrition, 2019, 31(6): 2854-2864.
LEE J, KIM M S.The role of GSK3 in glucose homeostasis and the development of insulin resistance[J].Diabetes Research and Clinical Practice, 2007, 77(Suppl.3):S49-S57.
XU H, ZHIOU Y, LIU Y X, et al.Metformin improves hepatic IRS2/PI3K/Akt signaling in insulin-resistant rats of NASH and cirrhosis[J].Journal of Endocrinology, 2016, 229(2):133-144.
WANG T, JIANG H M, CAO S J, et al.Baicalin and its metabolites suppresses gluconeogenesis through activation of AMPK or AKT in insulin resistant HepG-2 cells[J].European Journal of Medicinal Chemistry, 2017, 141:92-100.
[10]
PARK S A, CHOI M S, KIM M J, et al.Hypoglycemic and hypolipidemic action of Duzhong (Eucommia ulmoides Oliver) leaves water extract in C57BL/KsJ-db/db mice[J].Journal of Ethnopharmacology, 2006, 107(3):412-417.
[13]
CHENG Z Y, TSENG Y, WHITE M.Insulin signaling meets mitochondria in metabolism[J].Trends in Endocrinology & Metabolism, 2010, 21(10):589-598.
[2]
ZHOU J F, ZHANG T M, CHEN W A, et al.Comparative analysis of chemical components between barks and leaves of Eucommia ulmoides Oliver[J].Journal of Central South University, 2009, 16(3):371-379.
[9]
LEE M K, KIM M J, CHO S Y, et al.Hypoglycemic effect of Duzhong (Eucommia ulmoides Oliv.) leaves in streptozotocin-induced diabetic rats[J].Diabetes Research and Clinical Practice, 2005, 67(1):22-28.
[21]
WANG Q, WANG N, DONG M, et al.GdCl3 reduces hyperglycaemia through Akt/FoxO1-induced suppression of hepatic gluconeogenesis in Type 2 diabetic mice[J].Clinical Science, 2014, 127(2):91-100.
[23]
SAMUEL V T, SHULMAN G I.The pathogenesis of insulin resistance:integrating signaling pathways and substrate flux[J].Journal of Clinical Investigation, 2016, 126(1):12-22.
[3]
DO M H, HUR J, CHOI J, et al.Eucommia ulmoides ameliorates glucotoxicity by suppressing advanced glycation end-products in diabetic mice kidney[J].Nutrients, 2018, 10(3):265.
[7]
JIN X, AMITANI K, ZAMAMI Y, et al.Ameliorative effect of Eucommia ulmoides Oliv.leaves extract (ELE) on insulin resistance and abnormal perivascular innervation in fructose-drinking rats[J].Journal of Ethnopharmacology, 2010, 128(3):672-678.
[12]
CAI Y, WANG Y, ZHI F, et al.The effect of sanggua drink extract on insulin resistance through the PI3K/AKT signaling pathway[J].Evidence-Based Complementary and Alternative Medicine, 2018, 2018:9407945.
[14]
XIE W, YE Y L, FENG Y, et al.Linderane suppresses hepatic gluconeogenesis by inhibiting the cAMP/PKA/CREB pathway through indirect activation of PDE 3 via ERK/STAT3[J].Frontiers in Pharmacology, 2018, 9:476.
[17]
XIONG X P, SONG Q, HAN C C, et al.Insulin promotes the expression of the gluconeogenic rate-limiting enzymes phosphoenolpyruvate carboxykinase (Pepck) and glucose 6-phosphatase (G6pase) through PI3k/Akt/mTOR signaling pathway in goose hepatocytes[J].Revista Brasileira de Ciência Avícola, 2016, 18(3):395-400.
[18]
NAKAE J, KITAMURA T, SILVER D L, et al.The forkhead transcription factor Foxo1(Fkhr) confers insulin sensitivity onto glucose-6-phosphatase expression[J].Journal of Clinical Investigation, 2001, 108(9):1359-1367.
[6]
ZHANG Y Y, ZHANG H X, WANG F, et al.The ethanol extract of Eucommia ulmoides Oliv.leaves inhibits disaccharidase and glucose transport in Caco-2 cells[J].Journal of Ethnopharmacology, 2015, 163:99-105.
[20]
SHI Y N, QIAO J Y, MU B, et al.3-(2-amino-ethyl)-5-[3-(4-butoxyl-phenyl)-propylidene]-thiazolidine-2, 4-dione (K145) ameliorated dexamethasone induced hepatic gluconeogenesis through activation of Akt/FoxO1 pathway[J].Biochemical and Biophysical Research Communications, 2017, 493(1):286-290.
[22]
SAMUEL V T, CHOI C S, PHILLIPS T G, et al.Targeting FoxO1 in mice using antisense oligonucleotide improves hepatic and peripheral insulin action[J].Diabetes, 2006, 55(7):2042-2050.
[26]
LIU T Y, SHI C X, GAO R, et al.Irisin inhibits hepatic gluconeogenesis and increases glycogen synthesis via the PI3K/Akt pathway in type 2 diabetic mice and hepatocytes[J].Clinical Science, 2015, 129(10):839-850.
