Liu Shujie, Li Yongming, Deng Bo, Xu Ziwei
Institute of Animal Husbandry and Veterinary Science, Zhejiang Academy of Agricultural Sciences, 145 Shiqiao Road, Hangzhou, 310021, Zhejiang, China.
BMC Biotechnol. 2016 Mar 1;16:25. doi: 10.1186/s12896-016-0255-z.
Insulin-like growth factor I (IGF-I) is one important family of growth factors, which plays key role in intestinal growth, regeneration, and damage repair. However, the low natural abundance of IGF-I limits its research opportunities and practical application in the fields of medicine and animal husbandry. In this study, a tandem repeat strategy was used to express three copies of the same pIGF-I3 protein in L. lactis. The activity of recombinant pIGF-I3 (rpIGF-I3) was further examined by a mouse model of dextran sulfate sodium (DSS)-induced colitis. In addition, the potential of recombinant L. lactis expressing pIGF-I3 to reduce inflammatory disease was evaluated.
pIGF-I3 could be expressed in L. lactis by the detection of SDS-PAGE and Western blot. Experimental colitis was induced in BALB/c mice by administration of 5 % DSS in drinking water, and the clinical symptoms were observed in DSS-treated mice. Oral administration of recombinant L. lactis expressing pIGF-I3 improved the colonic architecture, and significantly reduced the increase of colonic damage score (P < 0.05). Furthermore, recombinant L. lactis expressing pIGF-I3 treatment significantly reduced serum DAO activity and colonic MPO level, and elevated colonic occludin level compared to the DSS group (P < 0.05).
The pIGF-I3 expressed in L. lactis has good biological activity, and oral administration of recombinant L. lactis expressing pIGF-I3 attenuated the symptoms and development of DSS-induced colitis in mice. These suggested that L. lactis could be a potential host bacterium for production and delivery of IGF-I against intestinal diseases.
胰岛素样生长因子I(IGF-I)是生长因子的一个重要家族,在肠道生长、再生和损伤修复中起关键作用。然而,IGF-I的天然丰度较低,限制了其在医学和畜牧领域的研究机会及实际应用。在本研究中,采用串联重复策略在乳酸乳球菌中表达三个相同的pIGF-I3蛋白拷贝。通过葡聚糖硫酸钠(DSS)诱导的小鼠结肠炎模型进一步检测重组pIGF-I3(rpIGF-I3)的活性。此外,评估了表达pIGF-I3的重组乳酸乳球菌减轻炎症性疾病的潜力。
通过SDS-PAGE和蛋白质免疫印迹检测发现pIGF-I3可在乳酸乳球菌中表达。通过在饮用水中给予5% DSS诱导BALB/c小鼠发生实验性结肠炎,并观察DSS处理小鼠的临床症状。口服表达pIGF-I3的重组乳酸乳球菌改善了结肠结构,并显著降低了结肠损伤评分的增加(P < 0.05)。此外,与DSS组相比,表达pIGF-I3的重组乳酸乳球菌治疗显著降低了血清二胺氧化酶(DAO)活性和结肠髓过氧化物酶(MPO)水平,并提高了结肠紧密连接蛋白(occludin)水平(P < 0.05)。
在乳酸乳球菌中表达的pIGF-I3具有良好的生物学活性,口服表达pIGF-I3的重组乳酸乳球菌可减轻小鼠DSS诱导的结肠炎的症状和发展。这些结果表明,乳酸乳球菌可能是生产和递送IGF-I以对抗肠道疾病的潜在宿主菌。