The Protection Of Lineage-Restricted Neural Precursors By Nourishing Piyin Remedy Against Lipid Peroxidation

Background: Spinal cord injury is a common multiple injury in department of orthopaedics,of which the clinical efficacy is not perfect. For the past few years,people realized that the permanent nerve injury of spinal cord is caused by some secondary responses after the primary injury of the cord. Some scholars presumed that spinal cord injury could be cured by cellular transplant after much research,and the NRPS cell was a good choise so far.But when people improved the method of cellular transplant,they found the part microenvironment was also a key factor for the transplant.Nourishng Piyin Remedy was invested for a long time in our college,we found it could protect the injured spinal cord of the rabbit and improve the ability of mitochondria membrane of cerebral nerves against the lipid peroxidation.It is a new topic that if Nourishng Piyin Remedy can elevat the survival of the transplanted cells,which is needed to be invested.Objective: To invest the protection of Lineage-Restricted Neural Precursors peroxidized by hydrogen dioxide(H2O2) by Nourishing Piyin Remedy, and to establish the foundation of the remedy to the spinal injury by explantation in vivo.Methods: To confirm the NRPS cells after the spinal cord of the embryo of SD rats by immunofluorescence identification in situ.Then,made the cells of spinal cord for adherent culture for 3-5days.To establish them of the NRPS cells by immunofluorescence identified by Nestin,PSA-NCAM,A2B5 antibodies . To make sure the component of blank-blood serum was different from the remedy-blood serum by infrared spectra identification.So we knew that the Nourishing Piyin remedy was in the blood serum.Establish the lipid peroxidation analogs for six groups:normal group,pure damage group,blank-blood serum protected group,remedy-blood serum protected group,damage after blank-blood serum protected group,damage after remedy-blood serum protected group.The concentrations of the blood serum were 0.25%,0.5%,1%,2%,4%,and the protection time was 8h;the concentrations of H2O2 were 0.05,0.1,0.2mmol/L,and the damage time was 4h.Observed the cells under the inverted phase contrast microscope to compare the cell morphology change; calculated out the cell survival rates by the MTT method; fluorescein stained the cells with hoechst33258, and then observed the fluor and the apoptotic body under the inverted fluorescence microscope;detected the content of MDA and the archaeus of SOD in cells to get the proper protecting concentration of the remedy-blood serum.Results: H2O2 cuted down the cell survival rate and speeds up the apoptosis noteblely. It enhanced the content of MDA and decreased the archaeus of SOD in cells. After the interfering of Nourishing Piyin Remedy at a certain concentration, the cell survival rates were raised at the same damage condition. But, the noxious property of Nourishing Piyin Remedy appeared when the concentration above 4%.Conclusion: Nourishing Piyin Remedy could protect the Lineage-Restricted Neural Precursors against lipid peroxidation at a certain concentration,can and the best concentration is 1%.It could prevent the decrease of SOD,and dimish the MOD of cells.