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分泌重组猪表皮生长因子的乳酸乳球菌可促进早期断奶仔猪的生长性能。

Recombinant porcine epidermal growth factor-secreting Lactococcus lactis promotes the growth performance of early-weaned piglets.

作者信息

Wang Dingyue, Xu Shengyu, Lin Yan, Fang Zhengfeng, Che Lianqiang, Xue Bai, Wu De

机构信息

Institute of Animal Nutrition, Key Laboratory for Animal Disease Resistance Nutrition of Sichuan Province, Sichuan Agricultural University, Ya'an 625014Sichuan, China.

出版信息

BMC Vet Res. 2014 Aug 21;10:171. doi: 10.1186/s12917-014-0171-1.

Abstract

BACKGROUND

Epidermal growth factor (EGF) is an important growth factor in regulation of cell proliferation, differentiation, survival and apoptosis. Studies showed that food-grade Lactococcus lactis (L. lactis) and NICE expression system have superior performance in exogenous protein expression. This study aimed to construct and express porcine EGF (pEGF), and use L. lactis as vehicle for producing and delivering pEGF. Furthermore, investigating biological activity of pEGF and exploring applications feasibility of combination effects of L. lactis and pEGF on early weaned piglets' production.

RESULTS

A recombinant Lactococcus lactis which produced and secreted pEGF at 1000 ng/ml in culture supernatant was generated. Secreted pEGF was a fully biologically active protein, as demonstrated by its capacity to stimulate L929 mouse fibroblast cell line proliferation in vitro. For in vivo study, forty piglets were randomly allocated to control, antibiotic control, empty vector-expressing L. lactis (LL-EV) and pEGF-secreting L. lactis (LL-pEGF). After 14 d of rearing, final body weight and average daily gain in LL-pEGF were greater (P < 0.05, 8.95 vs. 8.37 kg, 206.1 vs. 157.7 g/day, respectively) than those in control, but no significant differences between LL-pEGF, LL-EV and antibiotic control. Overall period average daily feed intake was higher in LL-pEGF, LL-EV and antibiotic control than in control (P < 0.05, 252.9, 255.6, 250.0, 207.3 g/day, respectively). No significant difference was observed on ADFI/ADG. LL-pEGF increased villous height in the duodenum, jejunum and ileum than in control and LL-EV (P < 0.05). Sucrase in the 3 intestinal segments, aminopeptidase A in the duodenum and Jejunum, aminopeptidase N and dipeptidase IV in the duodenum in LL-pEGF were higher than those in control (P < 0.05). Furthermore, Escherichia coli and Enterococcus counts decreased in the ileum and Lactobacillus increased in the ileum and cecum digesta in LL-pEGF compare with the control (P < 0.05). Lactobacillus increased in the cecum in LL-EV compared with control and antibiotic control (P < 0.05).

CONCLUSION

We have generated a recombinant Lactococcus lactis which produced and secreted fully biologically active porcine EGF. Oral administration of pEGF-secreting L. lactis had beneficial effects on intestinal health and performance of early-weaned piglets.

摘要

背景

表皮生长因子(EGF)是调节细胞增殖、分化、存活和凋亡的重要生长因子。研究表明,食品级乳酸乳球菌(L. lactis)和NICE表达系统在外源蛋白表达方面具有卓越性能。本研究旨在构建并表达猪表皮生长因子(pEGF),并利用乳酸乳球菌作为载体生产和递送pEGF。此外,研究pEGF的生物学活性,并探索乳酸乳球菌与pEGF联合作用对早期断奶仔猪生产性能的应用可行性。

结果

构建出一种重组乳酸乳球菌,其在培养上清液中产生并分泌的pEGF浓度为1000 ng/ml。分泌的pEGF是一种具有完全生物活性的蛋白质,体外刺激L929小鼠成纤维细胞系增殖的能力证明了这一点。在体内研究中,40头仔猪被随机分为对照组、抗生素对照组、表达空载体的乳酸乳球菌(LL-EV)组和分泌pEGF的乳酸乳球菌(LL-pEGF)组。饲养14天后,LL-pEGF组的末体重和平均日增重高于对照组(P < 0.05,分别为8.95 vs. 8.37 kg,206.1 vs. 157.7 g/天),但LL-pEGF组、LL-EV组和抗生素对照组之间无显著差异。LL-pEGF组、LL-EV组和抗生素对照组的全期平均日采食量高于对照组(P < 0.05,分别为252.9、255.6、250.0、207.3 g/天)。ADFI/ADG未观察到显著差异。LL-pEGF组十二指肠、空肠和回肠的绒毛高度高于对照组和LL-EV组(P < 0.05)。LL-pEGF组三个肠段的蔗糖酶、十二指肠和空肠的氨肽酶A、十二指肠的氨肽酶N和二肽酶IV均高于对照组(P < 0.05)。此外,与对照组相比,LL-pEGF组回肠中的大肠杆菌和肠球菌数量减少,回肠和盲肠内容物中的乳酸杆菌增加(P < 0.05)。与对照组和抗生素对照组相比,LL-EV组盲肠中的乳酸杆菌增加(P < 0.05)。

结论

我们构建出一种重组乳酸乳球菌,其能产生并分泌具有完全生物活性的猪表皮生长因子。口服分泌pEGF的乳酸乳球菌对早期断奶仔猪的肠道健康和生产性能具有有益影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c887/4236833/8676e4224211/s12917-014-0171-1-1.jpg

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