The bactericidal/permeability increasing protein (BPI) has an important function of nonspecific killing of Gram-negative bacteria. In this study, qPCR was used to detect the expression of the BPI gene in twelve tissue...The bactericidal/permeability increasing protein (BPI) has an important function of nonspecific killing of Gram-negative bacteria. In this study, qPCR was used to detect the expression of the BPI gene in twelve tissues of Meishan piglets from birth to weaning. BPI gene overexpression, bacterial adhesions count and indirect immunofluorescence were applied to analyze the relationship between BPI gene expression and the infectivity of Escherichia coli and Salmonella. The results showed that the BPI gene was expressed highly in duodenum, jejunum and ileum (fold changes of relative expression levels were more than 10 000, 500 and 200, respectively). The expression of the BPI gene at 35 days of age was significantly higher (P<0.01) than that at all other days. Transcription of the BPI gene was up-regulated 2 401-fold in porcine intestinal epithelial (IPEC-J2) cells that were transfected with the BPI gene overexpression lentivirus (IPEC-J2-BPI), and significantly higher (P<0.01) than that in negative control cells (IPEC-J2-NC). Protein expression levels in IPEC-J2-BPI cells were also increased. When IPEC-J2 cells were incubated with E. coli and Salmonella, respectively, for 2, 4, 6, 8, 10 and 12 h, the number of bacterial adhesions in IPEC-J2-BPI cells was significantly less (P<0.05) than that in IPEC-J2-NC cells. The results of indirect immunofluorescence analysis showed that the number of bacterial adhesions in IPEC-J2-BPI cells was significantly less (P<0.01) than that in IPEC-J2-NC cells. These results demonstrated that the BPI gene might play an important role in regulating weaning stress especially intestinal-mediated immune response. Overexpression of the BPI gene at the cellular level could significantly enhance the anti-bactericidal ability against Gram-negative bacteria such as E. coli and Salmonella. This has important biological significance in piglet resistance to bacterial diarrhea.展开更多
基金This study was supported by the National Natural Science Foundation of China(31772560)the Yangzhou University International Academic Exchange Fund(YZUIAEF201901005)+1 种基金the College Students’Innovation and Entrepreneurship Training Program of Jiangsu Province,China(201811117014Z)the Priority Academic Program Development(PAPD)of Jiangsu Higher Education Institutions.
文摘The bactericidal/permeability increasing protein (BPI) has an important function of nonspecific killing of Gram-negative bacteria. In this study, qPCR was used to detect the expression of the BPI gene in twelve tissues of Meishan piglets from birth to weaning. BPI gene overexpression, bacterial adhesions count and indirect immunofluorescence were applied to analyze the relationship between BPI gene expression and the infectivity of Escherichia coli and Salmonella. The results showed that the BPI gene was expressed highly in duodenum, jejunum and ileum (fold changes of relative expression levels were more than 10 000, 500 and 200, respectively). The expression of the BPI gene at 35 days of age was significantly higher (P<0.01) than that at all other days. Transcription of the BPI gene was up-regulated 2 401-fold in porcine intestinal epithelial (IPEC-J2) cells that were transfected with the BPI gene overexpression lentivirus (IPEC-J2-BPI), and significantly higher (P<0.01) than that in negative control cells (IPEC-J2-NC). Protein expression levels in IPEC-J2-BPI cells were also increased. When IPEC-J2 cells were incubated with E. coli and Salmonella, respectively, for 2, 4, 6, 8, 10 and 12 h, the number of bacterial adhesions in IPEC-J2-BPI cells was significantly less (P<0.05) than that in IPEC-J2-NC cells. The results of indirect immunofluorescence analysis showed that the number of bacterial adhesions in IPEC-J2-BPI cells was significantly less (P<0.01) than that in IPEC-J2-NC cells. These results demonstrated that the BPI gene might play an important role in regulating weaning stress especially intestinal-mediated immune response. Overexpression of the BPI gene at the cellular level could significantly enhance the anti-bactericidal ability against Gram-negative bacteria such as E. coli and Salmonella. This has important biological significance in piglet resistance to bacterial diarrhea.
