慢性重组人生长激素对小鼠肝脏STAT5的影响及其机制
|
摘要
目的研究慢性重组人生长激素(rhGH)刺激对幼鼠肝脏STAT5的影响及其分子机制。方法将32只幼鼠共分为4组:磷酸盐缓冲液(PBS)组、PBS+single rhGH组、chronic rhGH组、chronic rhGH+single rhGH组。慢性生长激素刺激组chronic rhGH和chronic rhGH+single rhGH两组共16只小鼠每天按照1μg/g的剂量腹腔注射rhGH,对照组和PBS+single rhGH组16只小鼠每天腹腔注射等体积PBS 0.1mol/L,共3周。注射完最后1次rhGH和PBS 16h后次日8:00,处死前30min PBS组和chronic rhGH组小鼠注射PBS 0.1mol/L,PBS+single rhGH和chronic rhGH+single rhGH组小鼠按1μg/g注射单剂量rhGH。结果慢性生长激素刺激组小鼠(chronic rhGH和chronic rhGH+single rhGH)与对照组小鼠(PBS组和PBS+single rhGH)相比体质量明显增加;与PBS组相比,chronic rhGH组小鼠肝脏GH信号通路内基础p-STAT5水平显著降低;凝胶迁移实验检测的chronic rhGH组小鼠肝脏内STAT5与DNA的结合能力显著低于PBS组;处死前30min按1μg/g单剂量注射rhGH的小鼠中,chronic rhGH+single rhGH组较PBS+single rhGH组小鼠肝脏p-STAT5水平则明显降低;chronic rhGH组与对照组相比,CIS含量显著升高,而SOCS-2的含量则无明显差异。结论 rhGH慢性刺激可抑制幼鼠肝脏细胞内STAT5的磷酸化,其机制可能与CIS含量的升高有关。
Objective To investigate the effect of chronic recombinant growth hormone(rhGH)stimulation on hepatic STAT5in healthy young mouse and its molecular mechanism.Methods 32normal young mice were divided into 4groups:PBS,PBS+single rhGH,chronic rhGH,chronic rhGH+single rhGH groups.The mice in the treatment groups(chronic rhGH and chronic rhGH+single rhGH)received intraperitoneal injection of rhGH at 1μg/g body weight once daily for three weeks;the mice in the control groups(PBS and PBS+single rhGH)were injected with sterile phosphate buffered saline(PBS,0.1mol/L).At next day 8:00after16hof injection of last rhGH and PBS,at 30min before killing mouse,PBS 0.1mol/L was injected in the PBS group and the chronic rhGH group,while the single dose of rhGH 1μg/g was injected in the PBS+single rhGH and the chronic rhGH+single rhGH group.Results Compared with the control mice(PBS group and PBS+single rhGH group),the body mass in the rhGH stimulation mice(chronic rhGH group and the chronic rhGH+single rhGH group)was significantly increased;compared with the PBS group,the base p-STAT5level within the hepatic GH signal pathway in the chronic rhGH was significantly decreased;the combining capacity of hepatic STAT5and DNA detected by EMSA in the chronic rhGH group was significantly lower than that in the PBS group;in the mice injected by single dose of rhGH 1μg/g at 30min before killing mouse,the hepatic p-STAT5level in the chronic rhGH+single rhGH was significantly decreased compared with the chronic rhGH+single rhGH;compared with the control group,the CIS content in the chronic rhGH was obviously increased,while the SOCS-2content had no obvious difference.Conclusion Chronic rhGH stimulation can inhibit the phosphorylation of hepatic STAT5in normal young mouse,which mechanism could be related with the increase of CIS content.
Objective To investigate the effect of chronic recombinant growth hormone(rhGH)stimulation on hepatic STAT5in healthy young mouse and its molecular mechanism.Methods 32normal young mice were divided into 4groups:PBS,PBS+single rhGH,chronic rhGH,chronic rhGH+single rhGH groups.The mice in the treatment groups(chronic rhGH and chronic rhGH+single rhGH)received intraperitoneal injection of rhGH at 1μg/g body weight once daily for three weeks;the mice in the control groups(PBS and PBS+single rhGH)were injected with sterile phosphate buffered saline(PBS,0.1mol/L).At next day 8:00after16hof injection of last rhGH and PBS,at 30min before killing mouse,PBS 0.1mol/L was injected in the PBS group and the chronic rhGH group,while the single dose of rhGH 1μg/g was injected in the PBS+single rhGH and the chronic rhGH+single rhGH group.Results Compared with the control mice(PBS group and PBS+single rhGH group),the body mass in the rhGH stimulation mice(chronic rhGH group and the chronic rhGH+single rhGH group)was significantly increased;compared with the PBS group,the base p-STAT5level within the hepatic GH signal pathway in the chronic rhGH was significantly decreased;the combining capacity of hepatic STAT5and DNA detected by EMSA in the chronic rhGH group was significantly lower than that in the PBS group;in the mice injected by single dose of rhGH 1μg/g at 30min before killing mouse,the hepatic p-STAT5level in the chronic rhGH+single rhGH was significantly decreased compared with the chronic rhGH+single rhGH;compared with the control group,the CIS content in the chronic rhGH was obviously increased,while the SOCS-2content had no obvious difference.Conclusion Chronic rhGH stimulation can inhibit the phosphorylation of hepatic STAT5in normal young mouse,which mechanism could be related with the increase of CIS content.
引文
[1]Kopchick JJ,Andry JM.Growth hormone(GH),GH receptor,and signal transduction[J].Mol Genet Metab,2000,71(2):293-314.
[2]Beyea JA,Sawicki G,Olson DM,et al.Hormone(GH)receptor knockout mice reveal actions of GH in lung development[J].Proteomics,2006,6(3):341-348.
[3]Spinola-Castro AM,Siviero-Miachon AA,da Silva MT,et al.The use of growth hormone to treat endocrine-metabolic disturbances in acquired immunodeficiency syndrome(AIDS)patients[J].Arq Bras Endocrinol Metabol,2008,52(5):818-832.
[4]Grunfeld C,Thompson M,Brown SJ,et al.Recombinant human growth hormone to treat HIV associated adipose redistribution syndrome 12week induction and 24week maintenance therapy[J].J Acquir Immune Defic Syndr,2007,45(3):286-297.
[5]Gupta V,Lee M.Growth hormone in chronic renal disease[J].Indian J Endocrinol Metab,2012,16(2):195-203.
[6]Youssef DM.Results of recombinant growth hormone treatment in children with end-stage renal disease on regular hemodialysis[J].Saudi J Kidney Dis Transpl,2012,23(4):755-764.
[7]Gebert CA,Park SH,Waxman DJ.Regulation of signal transducer and activator of transcription(STAT)5bactivation by the temporal pattern of growth hormone stimulation[J].Mol Endocrinol,1997,11(4):400-414.
[8]Gebert CA,Park SH,Waxman DJ.Down-regulation of liver JAK2-STAT5bsignaling by the female plasma pattern of continuous GH stimulation[J].Mol Endocrinol,1999,13(2):213-227.
[9]Choi HK,Waxman DJ.Growth hormone,but not prolactin,maintains low-level activation of STAT5a and STAT5bin female rat liver[J].Endocrinol,1999,140(11):5126-5135.
[10]Choi HK,Waxman DJ.Pulsatility of growth hormone(GH)signaling in liver cells:role of the JAK2/STAT5pathway in GH action[J].Growth Horm IGF Res,2000,10(1):1-8.
[11]王娟,高原,谭薇,等.人生长激素慢性刺激对小鼠肝细胞JAK/STAT5信号通路的影响[J].现代消化及介入诊疗,2011,16(1):28-31.
[12]Cicoira M,Kalra PR,Anker SD.Growth hormone resistance in chronic heart failure and its therapeutic implications[J].J Card Fail,2003,9(3):219-226.
[13]Rabkin R,Sun DF,Chen Y,et al.Growth hormone resistance in uremia,a role for impaired JAK/STAT signaling[J].Pediatr Nephrol,2005,20(3):313-318.
[14]Zhou Y,Xu BC,Maheshwari HG,et al.A mammalian model for Laron syndrome produced by targeted disruption of the mouse growth hormone receptor/binding protein gene(the Laron mouse)[J].Proc Natl Acad Sci USA,1997,94(24):13215-13220.
[15]DiFedele LM,He J,Bonkowski EL,et al.Tumor necrosis factor alpha blockade restores growth hormone signaling in murine colitis[J].Gastroenterology,2005,128(5):1278-1291.
[16]Greenhalgh CJ,Bertolino P,Asa SL,et al.Growth enhancement in suppressor of cytokine signaling 2(SOCS-2)-deficient mice is dependent on signal transducer and activator of transcription 5b(STAT5b)[J].Mol Endocrinol,2002,16(6):1394-1406.
[17]Schaefer F,Chen Y,Tsao T,et al.Impaired JAK-STAT signal transduction contributes to growth hormone resistance in chronic uremia[J].J Clin Invest,2001,108(3):467-475.
[18]Sun D,Zheng Z,Tummala P,et al.Chronic uremia attenuates growth hormone induced signal transduction in skeletal muscle[J].J Am Soc Nephrol,2004,15(10):2630-2636.
[19]Woelfle J,Rotwein P.In vivo regulation of growth hormone-stimulated gene transcription by STAT5b[J].Am J Physiol Endocrinol Metab,2004,286(3):393-401.
[20]Karlsson H,Gustafsson JA,Mode A.Cis desensitizes GH induced Stat5signaling in rat liver cells[J].Mol Cell Endocrinol,1999,154(1/2):37-43.
[21]Morales O,Faulds MH,Lindgren UJ,et al.1alfa,25-dihydroxyvitamin D3inhibits GH-induced expression of SOCS-3and CIS and prolongs growth hormone signaling via JAK2/STAT5system in osteoblast like cells[J].J Biol Chem,2002,277(38):34879-34884.
[22]Tollet-Egnell P,Flores-Morales A,Stavréus-Evers A,et al.Growth hormone regulation of SOCS-2,SOCS-3,and CIS messenger ribonucleic acid expression in the rat[J].Endocrinol,1999,140(8):3693-3704.
[2]Beyea JA,Sawicki G,Olson DM,et al.Hormone(GH)receptor knockout mice reveal actions of GH in lung development[J].Proteomics,2006,6(3):341-348.
[3]Spinola-Castro AM,Siviero-Miachon AA,da Silva MT,et al.The use of growth hormone to treat endocrine-metabolic disturbances in acquired immunodeficiency syndrome(AIDS)patients[J].Arq Bras Endocrinol Metabol,2008,52(5):818-832.
[4]Grunfeld C,Thompson M,Brown SJ,et al.Recombinant human growth hormone to treat HIV associated adipose redistribution syndrome 12week induction and 24week maintenance therapy[J].J Acquir Immune Defic Syndr,2007,45(3):286-297.
[5]Gupta V,Lee M.Growth hormone in chronic renal disease[J].Indian J Endocrinol Metab,2012,16(2):195-203.
[6]Youssef DM.Results of recombinant growth hormone treatment in children with end-stage renal disease on regular hemodialysis[J].Saudi J Kidney Dis Transpl,2012,23(4):755-764.
[7]Gebert CA,Park SH,Waxman DJ.Regulation of signal transducer and activator of transcription(STAT)5bactivation by the temporal pattern of growth hormone stimulation[J].Mol Endocrinol,1997,11(4):400-414.
[8]Gebert CA,Park SH,Waxman DJ.Down-regulation of liver JAK2-STAT5bsignaling by the female plasma pattern of continuous GH stimulation[J].Mol Endocrinol,1999,13(2):213-227.
[9]Choi HK,Waxman DJ.Growth hormone,but not prolactin,maintains low-level activation of STAT5a and STAT5bin female rat liver[J].Endocrinol,1999,140(11):5126-5135.
[10]Choi HK,Waxman DJ.Pulsatility of growth hormone(GH)signaling in liver cells:role of the JAK2/STAT5pathway in GH action[J].Growth Horm IGF Res,2000,10(1):1-8.
[11]王娟,高原,谭薇,等.人生长激素慢性刺激对小鼠肝细胞JAK/STAT5信号通路的影响[J].现代消化及介入诊疗,2011,16(1):28-31.
[12]Cicoira M,Kalra PR,Anker SD.Growth hormone resistance in chronic heart failure and its therapeutic implications[J].J Card Fail,2003,9(3):219-226.
[13]Rabkin R,Sun DF,Chen Y,et al.Growth hormone resistance in uremia,a role for impaired JAK/STAT signaling[J].Pediatr Nephrol,2005,20(3):313-318.
[14]Zhou Y,Xu BC,Maheshwari HG,et al.A mammalian model for Laron syndrome produced by targeted disruption of the mouse growth hormone receptor/binding protein gene(the Laron mouse)[J].Proc Natl Acad Sci USA,1997,94(24):13215-13220.
[15]DiFedele LM,He J,Bonkowski EL,et al.Tumor necrosis factor alpha blockade restores growth hormone signaling in murine colitis[J].Gastroenterology,2005,128(5):1278-1291.
[16]Greenhalgh CJ,Bertolino P,Asa SL,et al.Growth enhancement in suppressor of cytokine signaling 2(SOCS-2)-deficient mice is dependent on signal transducer and activator of transcription 5b(STAT5b)[J].Mol Endocrinol,2002,16(6):1394-1406.
[17]Schaefer F,Chen Y,Tsao T,et al.Impaired JAK-STAT signal transduction contributes to growth hormone resistance in chronic uremia[J].J Clin Invest,2001,108(3):467-475.
[18]Sun D,Zheng Z,Tummala P,et al.Chronic uremia attenuates growth hormone induced signal transduction in skeletal muscle[J].J Am Soc Nephrol,2004,15(10):2630-2636.
[19]Woelfle J,Rotwein P.In vivo regulation of growth hormone-stimulated gene transcription by STAT5b[J].Am J Physiol Endocrinol Metab,2004,286(3):393-401.
[20]Karlsson H,Gustafsson JA,Mode A.Cis desensitizes GH induced Stat5signaling in rat liver cells[J].Mol Cell Endocrinol,1999,154(1/2):37-43.
[21]Morales O,Faulds MH,Lindgren UJ,et al.1alfa,25-dihydroxyvitamin D3inhibits GH-induced expression of SOCS-3and CIS and prolongs growth hormone signaling via JAK2/STAT5system in osteoblast like cells[J].J Biol Chem,2002,277(38):34879-34884.
[22]Tollet-Egnell P,Flores-Morales A,Stavréus-Evers A,et al.Growth hormone regulation of SOCS-2,SOCS-3,and CIS messenger ribonucleic acid expression in the rat[J].Endocrinol,1999,140(8):3693-3704.