肝脏特异性含ZP结构域蛋白在肥胖小鼠模型中的表达
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摘要
目的·研究肝脏特异性含ZP结构域蛋白(liver-specific ZP domain-containing protein,LZP)在肥胖小鼠模型中的表达。方法·检测C57BL/6J小鼠各组织LZP的基因和蛋白表达水平。通过高脂饮食诱导C57BL/6J小鼠肥胖模型,以高脂饲料喂养ob/ob小鼠,在实验期间监测体质量和血糖,实验结束后检测肝质量和体脂,进行肝脏组织学检查。应用realtime-PCR检测肝脏组织LZP基因表达水平,Western blotting检测肝脏组织LZP蛋白表达水平。结果·Realtime-PCR结果表明LZP基因主要在C57BL/6J小鼠的肝脏表达,在脾脏和睾丸等部位表达量少;Western blotting结果表明仅在肝脏组织中检测到LZP蛋白表达。与普通饮食喂养的C57BL/6J小鼠相比,高脂饮食喂养的小鼠体质量、体脂和血糖增加(均P<0.05);肝质量增加(P<0.05),肝脏脂肪变明显;肝脏LZP在基因和蛋白表达水平均显著下降(均P<0.05)。与同窝野生型小鼠相比,ob/ob小鼠体质量和血糖均明显增高,但LZP在基因和蛋白水平均明显下降(均P<0.05)。结论·LZP在肥胖小鼠模型中表达明显下降,提示其可能参与机体的能量代谢。
Objective · To study the expression of liver-specific ZP domain-containing protein(LZP) in mouse models of obesity. Methods · The gene and protein expression of LZP in different tissues of C57 BL/6 J mice were detected by realtime-PCR and Western blotting respectively. C57 BL/6 J mice were treated with high fat diet(HFD) to establish the model of diet-induced obesity and ob/ob mice were also treated with HFD. The body mass and blood glucose were monitored during the experiment, then the liver weight and fat mass were measured at the end of the study. Hematoxylin-eosin staining of liver was performed to observe the morphology of liver. The expression of LZP in liver of model mice was also detected by realtime-PCR and Western blotting, respectively. Results · The expression of LZP m RNA was mainly found in liver, while a lower gene expression level was also observed in several other tissues such as spleen and testis by realtime-PCR. The protein expression of LZP was detected in liver in C57 BL/6 J mice by Western blotting.Compared with normal diet group, the group treated with HFD had significantly increased body mass and total fat mass, higher blood glucose, increased liver mass and more serious hepatic steatosis(all P<0.05), while the expression of LZP in liver was reduced(P<0.05). Similarly, body mass and blood glucose were increased significantly in ob/ob mice(both P<0.05), though the expression of LZP was decreased compared with wild type littermates(P<0.05). Conclusion · Mouse models of obesity display decreased expression of LZP in liver, indicating that LZP may play a role in metabolic homeostasis in obese individuals.
Objective · To study the expression of liver-specific ZP domain-containing protein(LZP) in mouse models of obesity. Methods · The gene and protein expression of LZP in different tissues of C57 BL/6 J mice were detected by realtime-PCR and Western blotting respectively. C57 BL/6 J mice were treated with high fat diet(HFD) to establish the model of diet-induced obesity and ob/ob mice were also treated with HFD. The body mass and blood glucose were monitored during the experiment, then the liver weight and fat mass were measured at the end of the study. Hematoxylin-eosin staining of liver was performed to observe the morphology of liver. The expression of LZP in liver of model mice was also detected by realtime-PCR and Western blotting, respectively. Results · The expression of LZP m RNA was mainly found in liver, while a lower gene expression level was also observed in several other tissues such as spleen and testis by realtime-PCR. The protein expression of LZP was detected in liver in C57 BL/6 J mice by Western blotting.Compared with normal diet group, the group treated with HFD had significantly increased body mass and total fat mass, higher blood glucose, increased liver mass and more serious hepatic steatosis(all P<0.05), while the expression of LZP in liver was reduced(P<0.05). Similarly, body mass and blood glucose were increased significantly in ob/ob mice(both P<0.05), though the expression of LZP was decreased compared with wild type littermates(P<0.05). Conclusion · Mouse models of obesity display decreased expression of LZP in liver, indicating that LZP may play a role in metabolic homeostasis in obese individuals.
引文
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[14]Petersen MC,Shulman GI.Mechanisms of insulin action and insulin resistance[J].Physiol Rev,2018,98(4):2133-2223.
[15]Han J,Li E,Chen L,et al.The CREB coactivator CRTC2 controls hepatic lipid metabolism by regulating SREBP1[J].Nature,2015,524(7564):243-246.
[16]Krycer JR,Sharpe LJ,Luu W,et al.The Akt-SREBP nexus:cell signaling meets lipid metabolism[J].Trends Endocrinol Metab,2010,21(5):268-276.
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[18]Lehrke M,Lazar MA.The many faces of PPARγ[J].Cell,2005,123(6):993-999.
[19]Ahmadian M,Suh JM,Hah N,et al.PPARγsignaling and metabolism:the good,the bad and the future[J].Nat Med,2013,19(5):557-566.
[20]Gross B,Pawlak M,Lefebvre P,et al.PPARs in obesity-induced T2DM,dyslipidaemia and NAFLD[J].Nat Rev Endocrinol,2017,13(1):36-49.
[21]Samuel VT,Shulman GI.Nonalcoholic fatty liver disease as a nexus of metabolic and hepatic diseases[J].Cell Metab,2018,27(1):22-41.
[2]NCD Risk Factor Collaboration.Trends in adult body-mass index in200 countries from 1975 to 2014:a pooled analysis of 1 698 populationbased measurement studies with 19.2 million participants[J].Lancet,2016,387(10026):1377-1396.
[3]Gross B,Pawlak M,Lefebvre P,et al.PPARs in obesity-induced T2DM,dyslipidaemia and NAFLD[J].Nat Rev Endocrinol,2017,3(1):36-49.
[4]Meex RCR,Watt MJ.Hepatokines:linking nonalcoholic fatty liver disease and insulin resistance[J].Nat Rev Endocrinol,2017,13(9):509-520.
[5]Xu ZG,Du JJ,Zhang X,et al.A novel liver-specific zona pellucida domain containing protein that is expressed rarely in hepatocellular carcinoma[J].Hepatology,2003,38(3):735-744.
[6]Wang CY,Liao JK.A mouse model of diet-induced obesity and insulin resistance[J].Methods Mol Biol,2012,821:421-433.
[7]Lutz TA,Woods SC.Overview of animal models of obesity[J].Curr Protoc Pharmacol,2012,Chapter 5:Unit5.61.
[8]Yang H,Wu C,Zhao S,et al.Identification and characterization of D8C,a novel domain present in liver-specific LZP,uromodulin and glycoprotein 2,mutated in familial juvenile hyperuricaemic nephropathy[J].FEBS Lett,2004,578(3):236-238.
[9]Fagerberg L,Hallstr?m BM,Oksvold P,et al.Analysis of the human tissuespecific expression by genome-wide integration of transcriptomics and antibody-based proteomics[J].Mol Cell Proteomics,2014,13(2):397-406.
[10]Shen HL,Xu ZG,Huang LY,et al.Liver-specific ZP domain-containing protein(LZP)as a new partner of Tamm-Horsfall protein harbors on renal tubules[J].Mol Cell Biochem,2009,321(1/2):73-83.
[11]Long J,Zhang ZB,Liu Z,et al.Loss of heterozygosity at the calcium regulation gene locus on chromosome 10q in human pancreatic cancer[J].Asian Pac JCancer Prev,2015,16(6):2489-2493.
[12]Lin HV,Accili D.Hormonal regulation of hepatic glucose production in health and disease[J].Cell Metab,2011,14(1):9-19.
[13]Altarejos JY,Montminy M.CREB and the CRTC co-activators:sensors for hormonal and metabolic signals[J].Nat Rev Mol Cell Biol,2011,12(3):141-151.
[14]Petersen MC,Shulman GI.Mechanisms of insulin action and insulin resistance[J].Physiol Rev,2018,98(4):2133-2223.
[15]Han J,Li E,Chen L,et al.The CREB coactivator CRTC2 controls hepatic lipid metabolism by regulating SREBP1[J].Nature,2015,524(7564):243-246.
[16]Krycer JR,Sharpe LJ,Luu W,et al.The Akt-SREBP nexus:cell signaling meets lipid metabolism[J].Trends Endocrinol Metab,2010,21(5):268-276.
[17]Shimano H,Sato R.SREBP-regulated lipid metabolism:convergent physiologydivergent pathophysiology[J].Nat Rev Endocrinol,2017,13(12):710-730.
[18]Lehrke M,Lazar MA.The many faces of PPARγ[J].Cell,2005,123(6):993-999.
[19]Ahmadian M,Suh JM,Hah N,et al.PPARγsignaling and metabolism:the good,the bad and the future[J].Nat Med,2013,19(5):557-566.
[20]Gross B,Pawlak M,Lefebvre P,et al.PPARs in obesity-induced T2DM,dyslipidaemia and NAFLD[J].Nat Rev Endocrinol,2017,13(1):36-49.
[21]Samuel VT,Shulman GI.Nonalcoholic fatty liver disease as a nexus of metabolic and hepatic diseases[J].Cell Metab,2018,27(1):22-41.