doi: 10.1186/s12896-016-0255-z.
Recombinant Lactococcus lactis expressing porcine insulin-like growth factor I ameliorates DSS-induced colitis in mice
Affiliations
- PMID: 26932768
- PMCID: PMC4774141
- DOI: 10.1186/s12896-016-0255-z
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Recombinant Lactococcus lactis expressing porcine insulin-like growth factor I ameliorates DSS-induced colitis in mice
BMC Biotechnol.
.
Abstract
Background: 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.
Results: 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).
Conclusions: 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.
Figures
Detection of rpIGF-I3 by SDS–PAGE (a) and Western blot (b) analysis of cell lysates of L. lactis NZ9000 (pNZ8148-pIGF-I3) induced with nisin. a Lanes 1 and 2, cell lysates of L. lactis NZ9000 (pNZ8148-pIGF-I3); lanes 3 and 4, cell lysates of L. lactis NZ9000 (pNZ8148); lane 5, molecular weight marker. b Lanes 1 and 3, cell lysates of L. lactis NZ9000 (pNZ8148); lanes 2 and 4, cell lysates of L. lactis NZ9000 (pNZ8148-pIGF-I3)
Effect of oral administration of L. lactis NZ9000 (pNZ8148-pIGF-I3) on colon length in mice (n = 8) treated with DSS-induced colitis. The statistical differences between groups were evaluated by one-way ANOVA with Duncan’s multiple comparison. Group 1 (control 
) received normal drinking water. Group 2 (DSS 
) received DSS solution in drinking water. Group 3 (L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148). Group 4 (recombinant L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148- pIGF-I3). Same letters between the groups indicate no statistically significant difference (P > 0.05), and different letters indicate statistically significant difference (P < 0.05)
) received normal drinking water. Group 2 (DSS ) received DSS solution in drinking water. Group 3 (L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148). Group 4 (recombinant L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148- pIGF-I3). Same letters between the groups indicate no statistically significant difference (P > 0.05), and different letters indicate statistically significant difference (P < 0.05)
Effect of oral administration of L. lactis NZ9000 (pNZ8148-pIGF-I3) on histopathological changes and colonic damage score in mice (n = 6) treated with DSS-induced colitis. The statistical differences between groups were evaluated by one-way ANOVA with Duncan’s multiple comparison. Group 1 (control 
) received normal drinking water. Group 2 (DSS 
) received DSS solution in drinking water. Group 3 (L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148). Group 4 (recombinant L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148-pIGF-I3). Same letters between the groups indicate no statistically significant difference (P > 0.05), and different letters indicate statistically significant difference (P < 0.05)
) received normal drinking water. Group 2 (DSS ) received DSS solution in drinking water. Group 3 (L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148). Group 4 (recombinant L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148-pIGF-I3). Same letters between the groups indicate no statistically significant difference (P > 0.05), and different letters indicate statistically significant difference (P < 0.05)
Effect of L. lactis NZ9000 (pNZ8148-pIGF-I3) on colonic MPO activity in mice (n = 6) treated with DSS-induced colitis. The statistical differences between groups were evaluated by one-way ANOVA with Duncan’s multiple comparison. Group 1 (control 
) received normal drinking water. Group 2 (DSS 
) received DSS solution in drinking water. Group 3 (L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148). Group 4 (recombinant L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148-pIGF-I3). Same letters between the groups indicate no statistically significant difference (P > 0.05), and different letters indicate statistically significant difference (P < 0.05)
) received normal drinking water. Group 2 (DSS ) received DSS solution in drinking water. Group 3 (L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148). Group 4 (recombinant L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148-pIGF-I3). Same letters between the groups indicate no statistically significant difference (P > 0.05), and different letters indicate statistically significant difference (P < 0.05)
Effect of L. lactis NZ9000 (pNZ8148-pIGF-I3) on serum DAO activity in mice (n = 6) treated with DSS-induced colitis. The statistical differences between groups were evaluated by one-way ANOVA with Duncan’s multiple comparison. Group 1 (control 
) received normal drinking water. Group 2 (DSS 
) received DSS solution in drinking water. Group 3 (L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148). Group 4 (recombinant L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148-pIGF-I3). Same letters between the groups indicate no statistically significant difference (P > 0.05), and different letters indicate statistically significant difference (P < 0.05)
) received normal drinking water. Group 2 (DSS ) received DSS solution in drinking water. Group 3 (L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148). Group 4 (recombinant L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148-pIGF-I3). Same letters between the groups indicate no statistically significant difference (P > 0.05), and different letters indicate statistically significant difference (P < 0.05)
Effect of L. lactis NZ9000 (pNZ8148-pIGF-I3) on colonic occludin level in mice (n = 6) treated with DSS-induced colitis. The statistical differences between groups were evaluated by one-way ANOVA with Duncan’s multiple comparison. Group 1 (control 
) received normal drinking water. Group 2 (DSS 
) received DSS solution in drinking water. Group 3 (L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148). Group 4 (recombinant L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148-pIGF-I3). Same letters between the groups indicate no statistically significant difference (P > 0.05), and different letters indicate statistically significant difference (P < 0.05)
) received normal drinking water. Group 2 (DSS ) received DSS solution in drinking water. Group 3 (L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148). Group 4 (recombinant L. lactis
) received the same treatment of DSS as Group 2. Meanwhile, these mice orally administered with L. lactis NZ9000 (pNZ8148-pIGF-I3). Same letters between the groups indicate no statistically significant difference (P > 0.05), and different letters indicate statistically significant difference (P < 0.05)Similar articles
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