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酵母表达的内源性表皮生长因子、外源性表皮生长因子和标记表皮生长因子对早期断奶仔猪生物学活性的比较。

Comparison of the biological activities of Saccharomyces cerevisiae-expressed intracellular EGF, extracellular EGF, and tagged EGF in early-weaned pigs.

机构信息

College of Life Science and Technology, Southwest University for Nationalities, Chengdu, 610041, People's Republic of China.

出版信息

Appl Microbiol Biotechnol. 2015 Sep;99(17):7125-35. doi: 10.1007/s00253-015-6468-6. Epub 2015 Feb 25.

Abstract

Epidermal growth factor (EGF) ameliorates stress and prevents incomplete gastrointestinal development in early-weaned piglets in commercial swine farming. This study aimed to further analyze the biological activities of intracellularly expressed EGF (IE-EGF), extracellularly expressed EGF (EE-EGF), and tagged EGF (T-EGF) from Saccharomyces cerevisiae in early-weaned pigs. In this study, we assigned 24 pigs to each of 5 groups that were provided a basic diet (the control group) or a diet supplemented with empty vector-expressing S. cerevisiae [the INVSc1(EV) group], T-EGF-expressing S. cerevisiae [the INVSc1-TE(-) group], EE-EGF-expressing S. cerevisiae [the INVSc1-EE(+) group], or IE-EGF-expressing S. cerevisiae [the INVSc1-IE(+) group]. All treatments were delivered at a dose of 60 μg EGF/kg body weight (BW) everyday. All the piglets were sacrificed after 21 day to determine their physio-biochemical indexes, immune functions, and intestinal development. In the piglet experiments, recombinant S. cerevisiae survived throughout the intestinal tract. The BW and intestinal development (e.g., mean villous height, crypt depth, villous height:crypt depth ratio (IVR), and total protein, DNA, and RNA contents) of the piglets were significantly enhanced in the INVSc1-IE(+) group compared with the animals in the INVSc1-EE(+) and INVSc1-TE(-) groups (P < 0.05). In addition, increased proliferating cell nuclear antigen (PCNA) staining was observed in the piglets that received the INVSc1-IE(+) treatment (approximately 80 %) compared with those that received the INVSc1-TE(-) (approximately 70 %) and INVSc1-EE(+) treatments (approximately 70 %). The levels of lactate dehydrogenase (LDH), creatine kinase (CK), alkaline phosphatase (ALP), immunoglobulin A (IgA), immunoglobulin M (IgM), and immunoglobulin G (IgG) were also significantly increased in the INVSc1-IE(+) group compared with the INVSc1-EE(+) and INVSc1-TE(-) groups (P < 0.05). Furthermore, the proliferation of piglet enterocytes was also significantly stimulated by both IE-EGF and EE-EGF compared with T-EGF in vitro (P < 0.05). Our data further demonstrate the previously reported hypothesis that IE-EGF is more suitable than EE-EGF or T-EGF for applications in early-weaned pigs.

摘要

表皮生长因子(EGF)可改善应激,预防商业养猪中早期断奶仔猪不完全胃肠道发育。本研究旨在进一步分析来自酿酒酵母的细胞内表达的表皮生长因子(IE-EGF)、细胞外表达的表皮生长因子(EE-EGF)和标记的表皮生长因子(T-EGF)在早期断奶仔猪中的生物学活性。在这项研究中,我们将 24 头猪分配到 5 组中,每组提供基础日粮(对照组)或添加表达空载体酿酒酵母的日粮[INVSc1(EV)组]、表达 T-EGF 的酿酒酵母[INVSc1-TE(-)组]、表达 EE-EGF 的酿酒酵母[INVSc1-EE(+)组]或表达 IE-EGF 的酿酒酵母[INVSc1-IE(+)组]。所有处理均以 60 μg EGF/kg 体重(BW)的剂量每天给药。所有仔猪在 21 天后处死,以确定其生理生化指标、免疫功能和肠道发育情况。在仔猪实验中,重组酿酒酵母在整个肠道中存活。与 INVSc1-EE(+)和 INVSc1-TE(-)组相比,INVSc1-IE(+)组仔猪的 BW 和肠道发育(如平均绒毛高度、隐窝深度、绒毛高度:隐窝深度比(IVR)以及总蛋白、DNA 和 RNA 含量)均显著提高(P < 0.05)。此外,接受 INVSc1-IE(+)治疗的仔猪中增殖细胞核抗原(PCNA)染色增加(约 80%),而接受 INVSc1-TE(-)和 INVSc1-EE(+)治疗的仔猪增加(约 70%)。与 INVSc1-EE(+)和 INVSc1-TE(-)组相比,INVSc1-IE(+)组的乳酸脱氢酶(LDH)、肌酸激酶(CK)、碱性磷酸酶(ALP)、免疫球蛋白 A(IgA)、免疫球蛋白 M(IgM)和免疫球蛋白 G(IgG)水平也显著升高(P < 0.05)。此外,与 T-EGF 相比,IE-EGF 和 EE-EGF 体外均显著刺激仔猪肠细胞增殖(P < 0.05)。我们的数据进一步证明了之前的假设,即 IE-EGF 比 EE-EGF 或 T-EGF 更适合应用于早期断奶仔猪。

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