促肾上腺皮质激素对母猪肝中葡萄糖代谢相关酶转录表达及活性的影响

doi:10.11843/j.issn.0366-6964.2018.11.005

摘要/Abstract

摘要：

旨在研究促肾上腺皮质激素（adrenocorticotropic hormone，ACTH）对母猪肝中葡萄糖代谢相关酶转录表达及活性的影响，以探讨应激状态下肝调节血糖水平的分子机制。本研究选取9头健康经产断奶苏淮母猪，随机分成处理组（5头）和对照组（4头）。对处理组母猪注射外源ACTH，对照组母猪注射生理盐水；每天处理3次，连续7 d。试验期间每天采集母猪血液；试验结束后，取母猪肝组织。利用放射免疫分析方法测定血液皮质醇浓度，利用生化分析仪测定血糖浓度，通过荧光定量PCR检测葡萄糖代谢相关酶和糖皮质激素受体（glucocorticoid receptor，GR）的转录表达，利用分光光度法检测葡萄糖代谢相关酶的酶活性。结果表明，ACTH处理组母猪血液中皮质醇及葡萄糖水平均显著高于对照组（P<0.05）。ACTH处理对母猪肝中丙酮酸羧化酶（PC）、果糖-1，6-二磷酸酶（FBP）、磷酸烯醇式丙酮酸羧激酶2（PCK2）、磷酸果糖激酶（PFKL）、丙酮酸激酶（PKLR）、己糖激酶1（HK1）、己糖激酶3（HK3）、糖原合成酶1（GYS1）、糖原磷酸化酶（PYGL）及6-磷酸葡萄糖脱氢酶（G6PD）的mRNA表达无显著影响（P>0.05）。ACTH处理显著上调母猪肝中葡萄糖-6-磷酸酶（G6PC）、磷酸烯醇式丙酮酸羧激酶1（PCK1）和己糖激酶2（HK2）的mRNA表达水平（P<0.05）。此外，ACTH处理显著上调母猪肝中葡萄糖-6-磷酸酶（G6PC）的酶活性（P<0.05），显著下调磷酸果糖激酶（PFKL）的酶活性（P<0.05）。与对照组相比，ACTH处理组GR mRNA表达水平呈上调趋势，但组间差异不显著（P>0.05）。结果提示，ACTH处理上调母猪血糖水平，不仅通过促进肝中糖异生关键酶G6PC、PCK1和HK2的转录表达及增强G6PC的活性增加糖异生反应，还通过降低糖酵解关键酶PFKL的活性抑制糖酵解反应。此外，ACTH处理未影响肝中GR的转录表达。

Abstract:

This study was designed to investigate the effects of adrenocorticotropic hormone (ACTH) on the gene transcription of glucose metabolism-related enzymes and related enzyme activities in the liver of sows, and to explore the molecular mechanism of blood glucose regulation in the liver under the stress condition. Nine healthy multiparous weaned Suhuai sows were selected and randomly divided into treatment group (N=5) and control group (N=4). Sows in treatment group and control group were injected with ACTH and physiological saline, respectively, three times daily for 7 days. The blood samples were collected daily during the experiment; The liver of sows were collected at the end of experiment. The concentrations of cortisol in blood were detected by radioimmunoassay. The levels of blood glucose were measured by biochemical analyzer. The transcript expression of enzymes related to glucose metabolism and glucocorticoid receptor(GR) were detected by real-time quantitative PCR, the activities of enzymes related to glucose metabolism were detected by spectrophotography. The results showed that the concentrations of cortisol and glucose in blood of sows were significantly raised after ACTH injection (P<0.05). ACTH treatment didn't significantly influence the mRNA expression levels of PC, FBP, PCK2, PEKL, PKLR, HK1, HK3, GYS1, PYGL and G6PD in the liver of sows (P>0.05). ACTH treatment significantly increased the mRNA expression levels of G6PC, PCK1 and HK2 in the liver of sows (P<0.05). Furthermore, ACTH treatment significantly increased the enzyme activities of G6PC (P<0.05), and significantly decreased the enzyme activities of PFKL (P<0.05). Compared with the control group,the mRNA expression levels of GR in the ACTH group showed an up-regulation trend, but there were no significant difference between the two groups (P>0.05). These results suggested that ACTH treatment upregulated blood glucose level through activating transcription of the key gluconeogenesis enzymes PCK1, HK2 and G6PC as well as the enzymatic activities of G6PC to promote gluconeogenesis, meanwhile, reducing the enzyme activities of the key glycolysis enzyme PFKL to repress the glycolysis in the liver. Furthermore, ACTH treatment didn't influence the mRNA expression of GR.

中图分类号:

孙晔, 曾梓菡, 贾璨灿, 宁彩波, 吴望军, 赵芳, 刘红林. 促肾上腺皮质激素对母猪肝中葡萄糖代谢相关酶转录表达及活性的影响[J]. 畜牧兽医学报, 2018, 49(11): 2340-2348.

SUN Ye, ZENG Zi-han, JIA Can-can, NING Cai-bo, WU Wang-jun, ZHAO Fang, LIU Hong-lin. The Effects of Adrenocorticotropic Hormone on the Transcript Expression and Activities of Glucose Metabolism-related Enzymes in the Liver of Sows[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(11): 2340-2348.

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