The Effect Of Cerebral Cortex Calreticulin In Surgical Traumatic Induced Neuroimmune Regulated By Electroacupuncture (EA) In Rats

Cairencuim is an ER(endopiasmic reticulum; luminai Ca2+Dinaing protein and molecular chaperon, which is expressed in all nucleated cells. Recently, it has documented that CRT play vital roles in regulation of immune response in several diseases, However, the neuroimmune modulation of CRT in response to stress or inflammation remains elusive. Accordingly, the present study determined to explore the involvement of CRT in EA immunomodulation in the traumatic rats.Method:(1)Three months (3-mon) and eighteen months (18-mon) male SD rats were performed with surgical traumatic stress. Bilateral EA of "Zusanli"(ST36) and "Sanyinjiao"(SP6) acupoints were stimulated. CCK-8was assayed for spleen lymphocyte proliferation and natural killer cells activity; Western Blotting, immunohistochemical analysis were used to detect CRT expression.(2) Immunofluorescence double staining and Confocal microscopy technique were used to detect subcellular distributing of in neuron, namely colocalization of CRT and CTB, CRT and c-Src.(3) In primary cortical neurons, after LPS exposure CRT distributing was detected by the Western-Blotting technique.(4) Central role of CRT in the neuroimmune modulation of EA was determined by ICV injection of CRT Phosphorat-hioate antisense oligodeoxy nucleotides (PS-ASODN).Result:(1) Surgical traumatic stress could induce immunosuppression in two age groups of rats,3-mon rats displayed deteriorated and prolonged immunosuppression than18-mon counterparts. EA could improve the immunosuppression in3-mon rats, but not in18-mon rats.(2)By immunofluorescence double staining, we have shown that CRT expression colocalized with the neuronal marker NeuN,but not with the microglia marker OX-42or the astrocyte marker glial fibrillary acidic Protein.(3) Surgical traumatic stress could induce the upregulation of CRT in the cerebral cortex.(4) By immunofluorescence double staining and Confocal microscopy technique,it revealed that the increased colocalization of CRT with c-Src and CTB in neurons after traumatic stress.(5) Expression of CRT and c-Src in the Primary cultured cortical neurons upregulated after LPS stimulation in one hour.(6) Immunosuppression could be reversed by ICV injection of CRT Phosphorat-hioate antisense oligodeoxy nucleotides (PS-ASODN), which mostly due to the downregulation of CRT. However, central modulation of CRT had no effect on the neuroimmune modulation of EA.Conclusion:(1) Surgical traumatic stress could induce immunosuppression with age depended manner.(2)In the cerebral cortex, CRT mainly expressed in neurons.(3) Surgical traumatic stress could induce the upregulation of CRT in the cerebral cortex, in parallel, distribution of CRT within neuron lipid rafts was strengthened.(4) CRT was involved in the neuroimmune modulation in traumatic rats. Taken together, the present results indicated that traumatic stress could induce CRT transolation from its orthotopic localization in the lumen of ER to the surface of the plasma membrane, by which neurons bestowed a capability to serve as a signal for antigen-presenting cells or target as "eat-me" signal for engulfment. However, EA might participate in neuroimmune modulation via other pathways.