Propionate enhances the expression of key genes involved in the gluconeogenic pathway in bovine intestinal epithelial cells
Propionate enhances the expression of key genes involved in the gluconeogenic pathway in bovine intestinal epithelial cells
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DOI：
10.3168/jds.2019-17309
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发表时间：
2020-06-01
影响因子：
3.5
通讯作者：
Zhao, Guo Qi
中科院分区：
文献类型：
--
作者：
Zhan, Kang;Yang, Tian Yu;Zhao, Guo Qi
大约15%到50%的短链脂肪酸(SCFA)到达反刍动物的小肠。以往的研究表明，丙酸诱导的小肠糖异生的激活对能量平衡有有益的影响。然而，丙酸对牛小肠肠上皮细胞中关键的糖异生基因的调节作用仍然鲜为人知。因此，本研究的目的是利用SV40T(1：200；上海圣克鲁斯，中国)建立牛空肠上皮细胞的长期培养体系，并探讨丙酸对牛空肠上皮细胞糖异生关键基因的调节作用。我们的研究表明，SV40T诱导的永生化可以建立长期的BIEC培养。永生化BIEC通过细胞角蛋白18、绒毛蛋白、脂肪酸结合蛋白2和小肠肽酶的表达来区分。与未经处理的对照组相比，20 mM的SCFA转运体、单羧酸转运体4和Na+/H+交换异构体1相关基因的mRNA表达显著增加。此外，BIEC还显著地从培养基中摄取丙酸和丁酸。值得注意的是，3 mM丙酸诱导了与糖异生有关的关键基因的mRNA水平的深刻变化，包括磷酸烯醇式丙酮酸羧基激酶2、丙酮酸羧基酶、果糖-1，6-二磷酸酶1和过氧化物体增殖物激活受体-伽玛共激活剂1α。此外，3 mM丙酸在孵育3、6、12和24 h时可促进PGC1a mRNA的表达。这些发现表明，丙酸控制着牛肠道细胞中涉及糖异生关键酶的基因的mRNA表达。
Approximately 15 to 50% of short-chain fatty acids (SCFA) reach the ruminant small intestine. Previous research suggests that activation of small intestinal gluconeogenesis induced by propionate has beneficial effects on energy homeostasis. However, the regulatory effect of propionate on key gluconeogenic genes in enterocytes of the bovine small intestine remains less known. Therefore, the purpose of this study was to establish the long-term cultures of bovine intestinal epithelial cells (BIEC) from bovine jejunum tissue using SV40T (1:200; Santa Cruz, Shanghai, China) and investigate the regulatory effect of propionate on the key gluconeogenic genes in BIEC. Our study showed that long-term BIEC cultures were established by SV40T-induced immortalization. Immortal BIEC were distinguished by the expression of cytokeratin 18, villin, fatty acid binding protein 2, and small intestine peptidase. The mRNA expression of genes involved in the SCFA transporters, monocarboxylate transporter 4, and Na+/H+ exchanger isoforms 1 were significantly elevated with 20 mM SCFA compared with untreated controls. In addition, BIEC exhibited significant uptake of propionate and butyrate from the culture medium. Remarkably, 3 mM propionate induced profound changes in mRNA level of key genes involved in gluconeogenesis, including phosphoenolpyruvate carboxykinase 2, pyruvate carboxylase, fructose-1,6-bisphosphatase 1, and peroxisome proliferator-activated receptor-gamma coactivator 1 alpha. Additionally, 3 mM propionate enhanced the expression of PGC1A mRNA at 3, 6, 12, and 24 h of incubation. These findings suggest that propionate controls the mRNA expression of genes involved in key enzymes for gluconeogenesis in the enterocytes of bovines.