Changes in Secretion and Phagocytosis of Rat Kupffer Cells Under Heat-stroke and Its Mechanism

CHEN Xiao-juan; HAO Qian; ZHU Guo-guo; LI Hui-min; LU Qi-ping

doi:10.13730/j.issn.1009-2595.2017.07.001

Abstract：Objective To investigate the secretion and phagocytosis changes of rat kupffer cells(KCs)under heat stroke(HS)and its mechanism.Methods The Wistar rats placed on body temperature maintenance were randomly assigned to normal control(NC)group and HS group.The rat livers of NC and HS groups were all dyed by hematoxylin-eosin(HE)staining,and the cytophagic situation of Indian ink in KCs were observed.All rat peripheral blood samples were collected before and after the experiment,endotoxin concentrations were detected by limulus test kit,and concerntrations of macrophage inflammatory protein-1 alpha(MIP-1α),tumor necrosis factor-alpha(TNF-α),interleukin-1 beta(IL-1β),interferon-gamma(INF-γ)were all tested by enzyme linked immunosorbent assay(ELISA).The paraffin sections were prepared,expressions of phosphorylation c-Jun N-terminal kinase(p-JNK)and MIP-1α in rat KCs were assessed by using immunofluorescence technique.The cell lysis buffer and Western-blot were used to extract total protein of liver tissues and detect the expressions of p-JNK as well as MIP-1α in rat KCs,respectively.Results HE staining results of rat liver showed phagocytosis with Indian ink of KCs in HS group was predominantly weaker than that of NC group(P<0.05).The concentrations of endotoxin in rat peripheral blood,MIP-1α,TNF-α,IL-1β and INF-γ in HS group were remarkably more than these of NC group(P<0.05).The expressions of p-JNK and MIP-1α were mainly expressed in KCs and liver by immunofluorescence.Compared with NC group,the expressions of p-JNK and MIP-1α in KCs of HS group obviously up-regulated(P<0.05).Conclusion The phagocytosis of HS rat decreses and secretory functions increase in inflammatory factors,such as TNF-α,IL-1β,INF-γ,etc.Its mechanism may relate to activate JNK signal path and grow up with MIP-1α expressions.