Altered beta-1,4-galactosyltransferase I expression during early inflammation after spinal cord contusion injury

作者:Niu Shuqiong; Fei Min; Cheng Chun; Yan Meijuan; Gao Shangfeng; Chen Mengling; Wang Haibo; Li Xin; Yu Xiaowei; Qian Ji; Qin Jing; Zhao Jian; Gu Jianxin; Shen Aiguo
来源:Journal of Chemical Neuroanatomy, 2008, 35(3): 245-256.
DOI:10.1016/j.jchemneu.2008.01.002

摘要

Post-traumatic inflammation has been implicated in secondary tissue damage after spinal cord injury (SCI). beta-1,4-Galactosyltransferase I (beta-1,4-GalT-I) is a key inflammatory mediator that plays a critical role in the initiation and maintenance of inflammatory reaction in diseases. The aim of the current study was to investigate whether beta-1,4-GaIT-I is expressed in SCI. Spinal cord contusion model was established in adult rats. Real-time PCR and Western blot analysis were used to detect the spatio-temporal expression of beta-1,4-GaIT-I after SCI Lectin-fluorescent staining with RCA-1 was used to detect the galactosylation of the membrane glycoproteins. The interaction and colocalization between beta-1,4-GalT-I and E-selectin in the injured spinal cords were also assessed by immunoprecipitation of E-selectin and double immunofluorescent staining, respectively. Real-time PCR revealed that beta-1,4-GalT-I mRNA reached the peak at 1 d after spinal cord contusion. In situ hybridization indicated that beta-1,4-GalT-I mRNA was mainly distributed in the local inflammatory cells, adjacent to the center of injury. Double immunofluorescent staining showed that beta-1,4-GalT-I mostly overlapped with ED 1-positive macrophages 1 d after SCI, partly colocalized with microglia, neutrophils and a few with oligodendrocytes and astrocytes. The result of Lectin-fluorescent staining with RCA-1 was similar to that of double immunofluorescent staining. Terminal galactosylation of E-selectin underwent obvious changes between sham and 3 d after SO by immunoprecipitation of E-selectin. Thus, the transient expression of high levels of beta-1,4-GalT-I may provide new insight into the early inflammation after SCI.