禁食与胰岛素处理对鳜血糖和leptin-A基因表达的影响（英文）
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摘要
为研究硬骨鱼类leptin基因表达与血糖之间的关系,研究在禁食与胰岛素处理后,检测鳜血糖和肝脏、肠道和脂肪组织leptin-A基因表达水平。在禁食实验中,鳜被禁食10d,并分别于禁食后0、4h、2d、6d和10d取样。禁食后6d鳜血糖开始降低,禁食后10d鳜血糖显著降低。同时,禁食后6d鳜肝脏leptin-A基因mRNA表达水平显著升高,禁食后4h鳜肠道和脂肪组织leptin-A基因mRNA表达水平均显著升高。在胰岛素处理实验中,分别于注射后12h和36h取样。腹腔注射胰岛素后12h,鳜血糖显著降低,而鳜肝脏、肠道和脂肪组织leptin-A基因mRNA表达水平无明显变化。腹腔注射insulin后36h,鳜肠道leptin-A基因mRNA表达水平显著升高。研究结果表明,长期禁食或胰岛素处理均能够降低鳜血糖水平,且影响消化器官和脂肪储存器官leptin-A基因表达。
Leptin has been reported to play important roles in regulation of glucose homeostasis in mammals.However,the relationship between blood glucose and leptin gene expression in teleosts is largely unknown.In the present study,blood glucose and leptin-A gene expression in liver,intestine and visceral adipose tissue were characterized in Chinese perch(Siniperca chuatsi) after fasting or insulin treatment. In the fasting experiment,Chinese perch were fasted for 10 days,sampling at 0,4 h,2 d,6 d and 10 d after fasting. Blood glucose was significantly decreased at 10 d after fasting; leptin-A mRNA expression was significantly increased in liver at 6 d after fasting,and significantly increased in intestine and visceral adipose tissue at 4 h after fasting.In the insulin treatment experiment,Chinese perch were intraperitoneally injected with bovine insulin(80 IU/kg body weight) or 0.7% saline as control,sampling at 12 h and 36 h post treatment. Blood glucose decreased significantly at 12 h after insulin treatment,and leptin-A mRNA expression in intestine increased significantly at 36 h after insulin treatment,which suggesting that prolonged fasting and insulin treatment decreased blood glucose,and increased leptin-A gene expression in fat depot organ and digestive organ of fish.
Leptin has been reported to play important roles in regulation of glucose homeostasis in mammals.However,the relationship between blood glucose and leptin gene expression in teleosts is largely unknown.In the present study,blood glucose and leptin-A gene expression in liver,intestine and visceral adipose tissue were characterized in Chinese perch(Siniperca chuatsi) after fasting or insulin treatment. In the fasting experiment,Chinese perch were fasted for 10 days,sampling at 0,4 h,2 d,6 d and 10 d after fasting. Blood glucose was significantly decreased at 10 d after fasting; leptin-A mRNA expression was significantly increased in liver at 6 d after fasting,and significantly increased in intestine and visceral adipose tissue at 4 h after fasting.In the insulin treatment experiment,Chinese perch were intraperitoneally injected with bovine insulin(80 IU/kg body weight) or 0.7% saline as control,sampling at 12 h and 36 h post treatment. Blood glucose decreased significantly at 12 h after insulin treatment,and leptin-A mRNA expression in intestine increased significantly at 36 h after insulin treatment,which suggesting that prolonged fasting and insulin treatment decreased blood glucose,and increased leptin-A gene expression in fat depot organ and digestive organ of fish.
引文
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[3]Vázquez M J,Romero-Ruiz A,Tena-Sempere M.Roles of leptin in reproduction,pregnancy and polycystic ovary syndrome:consensus knowledge and recent developments[J].Metabolism,2015,64(1):79-91
[4]Morton G J,Schwartz M W.Leptin and the central nervous system control of glucose metabolism[J].Physiological Reviews,2011,91(2):389
[5]Halaas J L,Gajiwala K S,Maffei M,et al.Weight-reducing effects of the plasma protein encoded by the obese gene[J].Science,1995,269(5223):543-546
[6]Fernández-Formoso G,Pérez-Sieira S,González-Touceda D,et al.Leptin,20 years of searching for glucose homeostasis[J].Life Sciences,2015,140:4-9
[7]Gorissen M,Bernier N J,Manuel R,et al.Recombinant human leptin attenuates stress axis activity in common carp(Cyprinus carpio L.)[J].General and Comparative Endocrinology,2012,178(1):75-81
[8]Lu R H,Zhou Y,Yuan X C,et al.Effects of glucose,insulin and triiodothyroxine on leptin and leptin receptor expression and the effects of leptin on activities of enzymes related to glucose metabolism in grass carp(Ctenopharyngodon idella)hepatocytes[J].Fish Physiology and Biochemistry,2015,41(4):981-989
[9]Aguilar A J,Conde-Sieira M,Polakof S,et al.Central leptin treatment modulates brain glucosensing function and peripheral energy metabolism of rainbow trout[J].Peptides,2010,31(6):1044-1054
[10]Kurokawa T,Uji S,Suzuki T.Identification of cDNA coding for a homologue to mammalian leptin from pufferfish Takifugu rubripes[J].Peptides,2005,26(5):745-750
[11]Huising M O,Geven E J,Kruiswijk C P,et al.Increased leptin expression in common carp(Cyprinus carpio)after food intake but not after fasting or feeding to satiation[J].Endocrinology,2006,147(12):5786-5797
[12]Murashita K,Uji S,Yamamoto T,et al.Production of recombinant leptin and its effects on food intake in rainbow trout(Oncorhynchus mykiss)[J].Comparative Biochemistry and Physiology B Biochemistry&Molecular Biology,2008,150(4):377-384
[13]Gorissen M,Bernier N J,Nabuurs S B,et al.Two divergent leptin paralogues in zebrafish(Danio rerio)that originate early in teleostean evolution[J].Journal of Endocrinology,2009,201(3):329-339
[14]Kurokawa T,Murashita K.Genomic characterization of multiple leptin genes and a leptin receptor gene in the Japanese medaka Oryzias latipes[J].General and Comparative Endocrinology,2009,161(2):229-237
[15]Won E T,Baltzegar D A,Picha M E,et al.Cloning and characterization of leptin in a Perciform fish the striped bass(Morone saxatilis):control of feeding and regulation by nutritional state[J].General and Comparative Endocrinology,2012,178(1):98-107
[16]Copeland D L,Duff R J,Liu Q,et al.Leptin in teleost fishes:an argument for comparative study[J].Frontiers in Physiology,2011,2:26
[17]Yuan X,Li A,Liang X F,et al.Leptin expression in mandarin fish Siniperca chuatsi(Basilewsky):regulation by postprandial and short-term fasting treatment[J].Comparative Biochemistry and Physiology Part A:Molecular&Integrative Physiology,2016,194:8-18
[18]He S,Liang XF,Li L,et al.Gene structure and expression of leptin in Chinese perch[J].General and Comparative Endocrinology,2013,194:183-188
[19]Livak K J,Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCTmethod[J].Methods,2001,25(4):402-408
[20]Leibson L,Plisetskaya E M.Effect of insulin on blood sugar level and glycogen content in organs of some cyclostomes and fish[J].General and Comparative Endocrinology,1968,11(2):381-392
[21]Parks W C,Drake R L.Insulin mediates the stimulation of pyruvate kinase by a dual mechanism[J].Biochemical Journal,1982,208(2):333-337
[22]Tinoco A B,Nisembaum L G,Isorna E,et al.Leptins and leptin receptor expression in the goldfish(Carassius auratus),Regulation by food intake and fasting/overfeeding conditions[J].Peptides,2012,34(2):329-335
[23]Suarez R K,Mommsen T P.Gluconeogenesis in teleost fishes[J].Canadian Journal of Zoology,1987,65(8):1869-1882
[24]Frühbeck G,Salvador J.Relation between leptin and the regulation of glucose metabolism[J].Diabetologia,2000,43(1):3-12