Regulation Of Nuclear HO-1 On The Permeability Of In Vitro Blood-spinal Cord Barrier After Hypoxia

ObjectiveTo study whether nuclear HO-1 reduce the permeability of in vitro blood-spinal cord barrier after hypoxia.MethodsCultured HCMEC/D3 and HA-sc,and HO-1C?23 Plasmid and NC plasmid were transfected into HCMEC/D3,three days after transfection with HO-1C?23,extract the nuclear and cytoplasmic proteins in HCMEC/D3 with HO-1C?23 and blank control group,western blot was used to detect the expression of HO-1protein in nuclear and cytoplasmic of HCMEC/D3 cells in HO-1C?23 group and blank control group.HCMEC/D3 and HA-sc use Transwell to construct in vitro blood-spinal cord barrier,the experiment was divided into HO-1C?23group?NC group?blank control group,each group was induced 24h by 150um of cocl₂.Western blot was used to detect the protein expression of caveolin-1?CAV-1??ZO-1?occludind and caveolin-2?CAV-2?.Immunofluorescence detection the distribution of caveolin-1?caveolin-2?ZO-1 and occludin.HRP was used to detect the permeability of blood spinal cord barrier.ResultsThe expression of HO-1 protein in nuclear in HO-1C?23 group was significantly higher than that in the blank control group?P<0.05?.However,there was no significant difference of HO-1 protein in cytoplasmic between HO-1C?23 group and blank control group?P>0.05?.After hypoxia,the expression of cav-1 and cav-2 protein in HO-1C?23 group was significantly decreased?P<0.05?.However,there was no significant difference in the expression of cav-1 and cav-2 protein between the NC group and blank control group?P>0.05?.Compared with NC group and blank control group,the expression of ZO-1 and occludin protein in HO-1C?23 group was significantly increased after hypoxia induction?P<0.05?.However,there was no significant difference in the expression of ZO-1 and occludin protein between the NC group and blank control group?P>0.05?.After hypoxia,the permeability of in vitro blood-spinal cord barrier transfected with HO-1C?23 group was significantly lower than that NC group and blank control group?P<0.05?.,compared with the blank control group,the permeability of blood-spinal cord barrier no significant difference?P>0.05?.Conclusions1 Nuclear HO-1 may reduces the permeability of the blood-spinal cord barrier by the transcellular route.2 Nuclear HO-1 may reduce the permeability of the blood-spinal cord barrier by the paracellular route.