Ginsenoside Rg1 Promote The Migration Of Olfactory Ensheathing Cells And Their Therapeutic Effects Of Spinal Cord Injury: An Experimental Study

Part1: Effect of ginsenoside Rg1 on migration of olfactory ensheathing cellObjective: To study the effects of ginsenoside Rg1 on migration of ensheathing olfactory cells(O ECs) as well as their expression of migration-related factors of matrix metalloproteinase-2(MMP-2), matrix metalloproteinase-9(MMP-9) and neural cell adhesion molecule 1(NCAM1).Methods : O ECs were primary cultured and purified from olfactory bulb of newborn SD rats. Purified cells were divided into 2 groups, the experimental group was treated with Rg1 in concentration of 40 μg/ml for 72 h(the optimal concentration and intervention time which have been proven by our previous study), the control group received an equal volume of DMEM/F12 medium, the migration of O ECs was examined by wound healing assay and transwell chamber assay. The expression of MMP-2, MMP-9 and NCAM1 m RNA was determined by RT-PCR. The expression of PI3K/Akt-related proteins is explored by Western-blot.Results: The results of wound healing assay show that more cells migrate across the scratch in experimental group(P<0.05), and transwell assay suggested that the Rg1 treated group had the larger number of OECs in the lower chamber than the control group(P<0.05). RT-PCR analysis demonstrated that Rg1 significantly up-regulated m RNA expression of MMP-2, MMP-9 and NCAM1(p<0.05). Ginsenoside Rg1 promoted the activity of PI3K/Akt signaling pathway.Conclusion: Ginsenoside Rg1 significantly promotes migration of OECs through up-regulating their m RNA expression of migration-related factors. Ginsenoside Rg1 promoted migration of OECs via PI3K/Akt signaling pathway.Part 2: Ginsenosides Rg1 enhance the ability of olfactory ensheathing cells to repair spinal cord injury: An Experimental StudyObjective: To study the effects of ginsenoside Rg1 on effects of ensheathing olfactory cells(OECs) in spinal cord injury as well as their expression of neurotrophic factors like ciliary neurotrophic factor(CNTF), vascular endothelial growth factor(VEGF).Methods : O ECs were primary cultured and purified from olfactory bulb of newborn SD rats. Purified cells were divided into 2 groups, the experimental group was treated with Rg1 in concentration of 40μg/ml for 72h(the optimal concentration and action time which have been proven by our previous study), the control group received an equal volume of DMEM / F12 medium. The models of spina l cord injury was established by using Allen’s combat, and were randomly divided into 3 groups, O ECs group(inject the control group cells), Rg1+O ECs group(inject the experimental group cells) and control group(inject an equal volume of DMEM / F12 medium). Using RT-PCR to determine m RNA expression of CNTF and VEGF in OECs group and Rg1+OECs group.Result: The BBB score of Rg1+O ECs group was higher than the other two groups(P<0.05). Histology revealed that Rg1+O ECs group had larger number of neurons, lower level of GFAP expression and less damage of spinal cord tissue than OECs group and control group. RT-PCR analysis demonstrated that Rg1 significantly up-regulated m RNA expression of CNTF and VEGF.Conclusion: Ginsinoside Rg1 obviously improve the effects of olfactory ensheathing cell transplantation for spinal cord injury by the up-regulation of CNTF and VEGF.