Study On Effects Of Microencapsulated Olfactory Ensheathing Cells Transplantation On Neuropathic Pain Mediated By P2X4 Receptors

Objective: To explore the microencapsulated olfactory ensheathing cells transplantation in the treatment of neuropathic rats,and to observe the dorsal root ganglion P2X4 receptors expression effects.Method: 1.The olfactory bulb tissue was taken from a SD rat head,using Nash differential adhesion method to culture the olfactory ensheathing cells and do the cell morphology and purity identification;2.Cultivate of microencapsulated olfactory ensheathing cells 3,96 healthy SD rats were randomly divided into four groups,n = 24,namely: the sham groups(sham groups);the groups of rat sciatic nerve(model groups=CCI);olfactory ensheathing cells transplantion on sciatic nerve constriction injury(OESCs groups);microencapsulated olfactory ensheathing cells transplantation on sciatic nerve constriction injury(MC-OESC groups);4,At 7,14 d.Using behavioral methods determination of rat mechanical withdrawl threshold and observe the expression level of P2X4 receptor in dorsal root ganglion of L4-5 using RT-PCR,immunohistochemistry staining and Western Blot technology.Result::At the 7th and 14 th days after operation:(1)Using behavioral methods determination of rat mechanical withdrawl threshold,compared with the sham groups,the CCI groups rats mechanical flinching reflex threshold was significantly reduced;compared with the CCI groups,OECs groups and MC-OECs groups rat mechanical flinching reflex threshold significantly increase;compared with the OECs groups,MC-OECs groups rat mechanical flinching reflex threshold is slightly increase.(2)At L4-5 dorsal root ganglion P2X4 receptors RT-PCR m RNA expression levels in CCI groups compared with the sham groups,m RNA expression have significantly increased;but compared with the CCI groups,olfactory ensheathing cells groups and microencapsulated olfactory ensheathing cells groups was significantly reduced.(3)Immunohistochemistry: compared with the sham groups,L4-5 dorsal root ganglion P2X4 receptors average absorbance have significantly increased in CCI groups while compare with CCI group,OECs groups and MC-OECs groups were significantly lower.(4)Western blot: compared with sham groups,CCI groups P2X4 receptors protein expression was significantly increased,while compared with CCI groups,OECs groups and MC-OECs groups were significantly reduced at L4-5 dorsal root ganglion.,compared with the OECs groups,the MC-OECs groups in the L4-5 dorsal root ganglion P2X4 receptor m RNA,the average absorbance and protein expression level was slightly lower than that of the OECs groups.Conclusion: Olfactory ensheathing cells and microencapsulated olfactory ensheathing cells transplantation can decrease neuropathic pain after sciatic nerve injury,it mybe retated to reduce P2X4 receptors expression levels at the L4-5 dorsal root ganglion,the treatment effects of microencapsulated olfactory ensheathing cells groups were better than that of olfactory ensheathing cells groups.