分子营养 Molecular Nutrition

表皮生长因子调控猪肠上皮细胞中钠依赖Ⅱb型磷转运蛋白表达的细胞信号通路研究

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  • 湖南农业大学动物科学技术学院, 湖南畜禽安全生产协同创新中心, 长沙 410128
邢廷杰(1981-),男,湖北阳新人,博士研究生,从事动物营养与饲料研究。E-mail:xingtingjie@dbn.com.cn

收稿日期: 2016-12-20

  网络出版日期: 2017-06-07

基金资助

国家自然科学基金面上项目(31572419);长沙市科技计划项目(kh1601185)

Identifying Cell Signaling Pathway that Mediates Epidermal Growth Factor Regulating Sodium Dependent Phosphate Cotransporters Type Ⅱb Expression in Porcine Intestinal Epithelial Cells

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  • College of Animal Science and Technology, Hunan Agricultural University, Hunan Co-Innovation Center of Animal Production Safety, Changsha 410128, China

Received date: 2016-12-20

  Online published: 2017-06-07

摘要

本研究旨在探讨表皮生长因子(EGF)调控猪小肠上皮细胞IPEC-J2中钠依赖Ⅱb型磷转运蛋白(NaPi-Ⅱb)表达的分子机制。试验分别用EGF受体酪氨酸激酶抑制剂(tyrphostin AG 1478)、蛋白激酶A(PKA)抑制剂(H89)、蛋白激酶C(PKC)抑制剂(k4393)、p38抑制剂(SB 203580)、细胞外信号调节激酶(ERK)抑制剂(PD98059)、c-Jun氨基末端激酶(JNK)抑制剂(anisomycin)与EGF共同处理IPEC-J2细胞,利用Western blot检测相关通路蛋白及目的蛋白(NaPi-Ⅱb)的表达水平。结果显示:相较于对照组,EGF处理后NaPi-Ⅱb表达水平显著降低(P<0.05);相较于无抑制剂组,EGF受体、PKA、PKC、丝裂原活化蛋白激酶(MAPK)/p38、MAPK/ERK1/2、MAPK/JNK的特异性抑制剂处理IPEC-J2后,NaPi-Ⅱb表达水平显著提高(P<0.05),其中添加MAPK/ERK1/2特异性抑制剂显著降低了MAPK/ERK1/2在Tyr204位点的磷酸化水平(P<0.05),添加MAPK/JNK的特异性抑制剂显著降低了MAPK/JNK1/2/3在Thr183和Tyr185位点的磷酸化水平(P<0.05),说明该2组抑制剂对该通路的抑制作用是通过降低上述位点的磷酸化水平实现的。本研究结果表明EGF受体、PKA、PKC、p38、ERK和JNK均介导了EGF调控IPEC-J2细胞中NaPi-Ⅱb的表达。

本文引用格式

邢廷杰, 汤小朋, 曹满湖, 方热军 . 表皮生长因子调控猪肠上皮细胞中钠依赖Ⅱb型磷转运蛋白表达的细胞信号通路研究[J]. 动物营养学报, 2017 , 29(6) : 1988 -1995 . DOI: 10.3969/j.issn.1006-267x.2017.06.020

Abstract

The aim of this study was to elucidate the molecular mechanisms of epidermal growth factor (EGF) down-regulated sodium dependent phosphate cotransporters type Ⅱb (NaPi-Ⅱb) expression in porcine intestinal epithelial cells (IPEC-J2). EGF receptor tyrosine kinase inhibitor (tyrphostin AG 1478), protein kinase A (PKA) inhibitor (H89), protein kinase C (PKC) inhibitor (k4393), p38 inhibitor (SB 203580), extracellular signal regulated kinase (ERK) inhibitor (PD98059) and c-Jun n-terminal kinase (JNK) inhibitor (anisomycin) as well as EGF were used to treat IPEC-J2 cells, respectively, and related signal pathways protein and interest protein (NaPi-Ⅱb) expression levels were determined by Western blot. The results showed as follows, compared with control group, NaPi-Ⅱb expression level was significantly decreased after EGF treatment (P<0.05). Compared with none inhibitor treatments, inhibitors of EGF receptor, PKA, PKC, mitogen activated protein kinase (MAPK)/p38, MAPK/ERK1/2 and MAPK/JNK significantly increased NaPi-Ⅱb expression (P<0.05). Especially, MAPK/ERK1/2 inhibitor significantly reduced the phosphorylation level of MAPK/ERK1/2 at Tyr204 site (P<0.05), and MAPK/JNK inhibitor significantly reduced the phosphorylation level of MAPK/JNK1/2/3 at Thr183 and Tyr185 sites (P<0.05). In summary, all EGF receptor, PKA, PKC, p38, ERK and JNK participate in the inhibitory effects of EGF on NaPi-Ⅱb expression.

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