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Objective:To explore the neuroprotective effect and mechanism of Buyang Huanwu Tang (BYHWT) on experimental autoimmune encephalomyelitis (EAE) at different stages. Method:The 36 female C57BL/6 mice were immunized subcutaneously with myelin oligodendrocyte glycoprotein peptides (MOG35-55),then randomly divided into 9, 17, 28 d EAE control group. Each BYHWT group was orally given drugs on the 3rd day after immunization (50 g·kg-1·d-1), and EAE control group was given the same volume of normal saline in the same way once a day for 9, 17 and 28 d after immunization. The effect of BYHWT on EAE mice was observed with internationally accepted clinical score. Brain and spinal cord specimens were collected at 9, 17 and 28 d after immunization. The neuroprotective effect of BYHWT was observed by hematoxylin-eosin(HE)staining and solid blue staining (LFB). The expressions of BDNF and GAP-43 in spinal cord and brain were detected by Western blot. Result:After treatment, BYHWT can significantly inhibit myelitis cell infiltration and alleviate myelin loss. Compared with EAE group, the expression of Nogo-A in the spinal cord of each BYHWT group was significantly down-regulated (PPPPConclusion:BYHWT can improve the local nerve growth microenvironment and promote the expression of NTFs, reduce the expressions of neuroinhibitory factors, and play a role in neuroprotection.
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Objective To evaluate the changes in the expression of Nogo-A protein in the dorsal root ganglion (DRG) and spinal dorsal horn in a rat model of inflammatory pain.Methods One hundred and twenty male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 2 groups (n =60 each):control group (group C) and formalin group (group F).The inflammatory pain was induced by injection of 3％ formalin 100 μl into the plantar surface of left hindpaw in group F.The equal volume of normal saline was given instead of formalin in group C.Mechanical withdrawal threshold to yon Frey filament stimulation was measured at 1 h and 1,2,3 and 7 days after injection.Twelve rats in each group were chosen at each time point and sacrificed.The L5 DRG and L4,5 segment of spinal cord on the operated side were removed for determination of Nogo-A protein expression by immunofluorescence and Western blot.Remlts Compared with group C,the mechanical withdrawal threshold was significantly decreased at 1 h,1,2,3 and 7 days after injection,and the expression of Nogo-A protein in the DRG and L4,5 segment of spinal cord was up-regulated at 1 h and 2,3 and 7 days after injection in group F (P ＜0.05).Conclusion Up-regulation of Nogo-A protein in the DRG and spinal dorsal horn may play an important role in the development of formalin-induced inflammatory pain in rats.
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Objective To study the effects of rehabilitation training combined with the transplantation of bone marrow mesenchymal stem cells-derived neural stem cells (BMSC-D-NSCs) on the expression of Nogo-A and NgR protein in rats after spinal cord injury (SCI).Methods The spinal cords of eighty Sprague-Dawley rats were injured using a modified Allen′s impactor (H = 25 mm) at T10. The injured rats were randomly divided into a combination therapy group which was given rehabilitation training and cell transplants, a cell graft group, a rehabilitation training group and a control group. At the 7th day post SCI, BMSC-D-NSCs were transplanted into the injured spinal cords of the rats in the combination therapy and cell graft groups. Hindlimb movement was assessed using the BassoBeattie-Bresnahan (BBB) scale every week, and protein was extracted from the injured spinal cord tissue for Nogo-A and NgR determination by Western blotting at the 1st, 3rd and 7th day after cell transplantation.Results The average BBB score of the rats in the combination therapy group was significantly higher than that of the other groups from 2 weeks post transplantation. The scores in the rehabilitation training group were significantly higher than in the control group from the 5th week post transplantation. Western blotting showed high expression of Nogo-A and NgR protein 24 h post surgery, but these declined with time. For Nogo-A there was a significant difference among the groups at all three time points. In the combination therapy group the expression declined to a minimum by the 7th day. For NgR protein there was no significant difference between the 1st and 3rd day in any group.Conclusions Rehabilitation training combined with BMSC-D-NSC transplantation can have a synergistic effect on functional recovery from SCI. It can down regulate the expression of Nogo-A and NgR protein.
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Objective To investigate the changes in the expression of Nogo-A protein in the dorsal root ganglion (DRG) and spinal dorsal horn in a rat model of neuropathic pain (NP) .Methods Seventy-two male SD rats weighing 250-300 g were randomly divided into 3 groups ( n = 24 each) : control group (group C) , sham operation group (group S) and NP group. NP was induced by ligation and severance of tibial and common fibular nerves according to the technique described by Isabelle et al. The mechanical withdrawal threshold (MWT) to von Frey filament stimulation was measured at 1, 7, 14 and 21 days after ligation. Six rats in each group were randomly selected at each time point and sacrificed (3 for determination of Nogo-A protein expression by immunofluorescence, 3 for determination of Nogo-A protein expression by Western blot) . The L5 DRG and L4,5 segment of spinal cord on the injured side were removed for determination of Nogo-A protein expression by immunofluorescence and Western blot. Results Compared with the groups C and S, MWT was significantly decreased at 7, 14 and 21 days after ligation, the expression of Nogo-A protein in the DRG was down-regulated at 7 and 14 days after ligation and the expression of Nogo-A protein in the spinal dorsal horn was up-regulated at 14 and 21 days after ligation ( P ＜0.05) .Conclusion The Nogo-A protein in the DRG and spinal dorsal horn may play an important role in peripheral nerve injury-induced NP in rats.