ABSTRACT
Objective To prepare and identify rabbit polyclonal antibody against embryonic liver fordrin 3(ELF3),and investigate the distribution of ELF3 in mice tissue.Methods ELF3 specific N-terminal peptide was synthesized,and conjugated to Keyhole limpet hemocyanin(KLH)as immunogen.The ELF3-KLH complex was injected into rabbits subcutaneously,and then ELF3 antibody was purified using affinity chromatography.The titer of the antibody was evaluated by ELISA.The specificity of antibody against ELF3 and immunolocalization of ELF3 were evaluated by using Western blot and immunohistochemistry.Results Rabbit polyclonal antibody against ELF3 was prepared by the immunization of ELF3-KLH complex.ELISA and Western blot results showed the antibody against ELF3 had high titer and specificity.Western blot and immunohistochemical studies demonstrated ELF3 was expressed in the mouse heart,liver,brain and kidney tissue,particular on the cell membrane.Conclusion The preparation of polyclonal antibody against ELF3 was successful due to its high titer and specificity;ELF3 was expressed in the mice heart,liver,and kidney,particular on the cell membrane.It will provide an excellent tool for further study on the ELF3 function.
ABSTRACT
To investigate the role of endothelial cell derived cytokines in sepsis induced liver and lung injury. Sepsis was induced in mice by cecal ligation and puncture(CLP). Sham operation group underwent the same manipulation but without CLP. The gene expressions of pro inflammatory cytokines (TNF?, IL 1? and IL 6) in pulmonary and hepatic microvascular endothelial cells were assessed by RT PCR. The results showed that TNF? and IL 1? mRNA levels markedly increased at 3h after CLP in both pulmonary and hepatic microvascular endothelial cells. Though at 12h after CLP both of them declined, the levels were still higher than those in sham group. IL 6 mRNA expression also increased significantly at 3h after CLP in both pulmonary and hepatic microvascular endothelial cells. At 12h after CLP, the expression of IL 6 mRNA in hepatic microvascular endothelial cells remained at a high level, while the level in pulmonary microvascular endothelial cells became higher. It suggested that endothelium is an important source of pro inflammatory cytokines in the liver and lung tissues during sepsis.