| [Objective]Observe the effects of Bo’fa method on the hyperalgesia of the chronic constriction injury(CCI)model rats through the behavioral and morphology.Then study the changes of the expression of the central inhibitory neurotransmitter 5-HT2A and GABA-B receptors,the proinflammatory cytokine IL-1β,and the IL-1β in peripheral serum,to explore the effect mechanism of Neuropathic Pain intervened by Bo’fa method,and perfect the mechanism of Bo’fa method promoting the repair of sciatic nerve injury.This research will provide the forceful theoretical evidence for the clinical application of Bo’fa method treating the Neuropathic Pain related diseases,improving and noticing in advance the patients’ related symptoms.[Method]As the experimental animal,SD rats were randomly divided into four groups,including the normal group,the fake operation group,the Bo’fa method group and the model group.The CCI models were caused by the ligating round the sciatic nerve in the Bo’fa method group and the model group.The rats in the Bo’fa method group were intervened by the simulant Bo’fa method of Massage and Tuina Simulation Instrument.They were intervened qualitatively and quantitatively by stimulating orderly the acupoints of Yinmen(BL37),Chengshan(BL57)and Yanglingquan(GB34)with the length of 3 minutes per acupoint and method.After the intervention,to study the influence of Bo’fa method on the sensory function recovery of Neuropathic Pain in three terms below:1.The 7th day after modeling and the moment after 20 days of Bo’fa method treatment were chosen as the two time points.By the experiment of the solar-thermal pain threshold to observe the thermohyperalgesia condition of rats,we explored the behavioral evidence of the Bo’fa method alleviating the hyperalgesia condition of Neuropathic Pain.Use the light microscope to observe and analyze the ultrastructure changes of the axon and myelin sheath in sciatic nerve injury,to further explore the morphological evidenc.e of Bo’fa method promoting the injury recovery of Neuropathic Pain.2.Through the method of Western blot to analyze the protein quantitatively of the 5-HT2A receptor in L3-L5 segments of spinal cord,and the immunohistochemistry method to analyze the GABA-B receptor in L3-L5 segments of spinal cord,we explored whether Bo’fa method could promote the expression of the central inhibitory neurotransmitter 5-HT2A and GABA-B receptors,and alleviate the derepression of the central system,to inhibit the process of the Neuropathic Pain occurring and developing and relieve the Neuropathic Pain related symptons effectively.3.To further explore the mechanism of Bo’fa method promoting the inflammatory pain,we analyzed quantitatively the level of the proinflammatory cytokine IL-1β in rat serum by the method of enzyme linked immunosorbent assay(ELISA),and made the locational analysis about IL-1β in spinal dorsal horn with the immunohistochemistry method.To explore whether Bo’fa method intervene could decrease the expression of inflammatory factors in spinal dorsal horn and peripheral serum effectively.Through eliminating the inflammation,Bo’ fa method could reduce the central sensitization in the central system,and reduce the inflammatory sysmtoms of Neuropathic Pain in the peripheral system.[Results]1.The behavior results:After modeling 7 days,the behavior performances of the Bo’fa method group and the model group were the same(P>0.05).The solar-thermal pain threshold of that two groups were significantly lower than the normal group(P<0.05),and that in the fake operation group was the same with the normal group(P>0.05).After 20 days of Bo’fa method intervene,the solar-thermal pain threshold of the model group and the Bo’fa method group both decreased significantly,compared with the normal group(P<0.05),and that in the Bo’fa method group increased obviously,compared with the model group(P<0.05).2.The morphological results:After modeling 7 days,in the normal group and the fake operation group,it was visible that the sciatic neuronal axons arranged neatly,with myelin sheath in the complete structure.In the model group,the sciatic neuronal axons cracked,and myelin sheath was broken.It was visible for the proliferation of a great number of broken Schwann cells.After 20 days of Bo’fa method intervene,in the model group,the sciatic neuronal axons and myelin sheath slightly arranged in order.And it was still visible for the proliferation of broken Schwann cells.Compared with the model group,in the Bo’fa method group the arrangement of nerve fibers was more slightly ordered,the sciatic neuronal axons were blurred visible,and the proliferation of Schwann cells was reduced obviously.3.The results of the expression of 5-HT2A and GABA-B receptors in spinal cord:After modeling 7 days,the protein expression of 5-HT2A and GABA-B receptors in spinal cord of CCI model rats in the Bo’fa method group and the model group were reduced obviously compared to the normal group(P<0.05).And there was no significant difference in that between the fake operation group and the normal group(P>0.05).After 20 days of Bo’fa method intervene,there was no significant difference in the protein expression of 5-HT2A and GABA-B receptors in rat spinal cord between the Bo’fa method group and the normal group(P>0.05).Compared with the normal group,that in the model group still reduced obviously(P<0.05).4.The results of expression of the central and peripheral IL-1β:After modeling 7 days,compared with the normal group,the expression of IL-1β in spinal dorsal horn and peripheral serum in the Bo’fa method group and the model group increased statistically(P<0.05).There was no statistical difference in that between the fake operation group and the normal group(P>0.05).After 20 days of Bo’fa method intervene,compared with the model group,that in the Bo’fa method group reduced obviously(P<0.05).The research results in this experiment showed that,after 20 days of Bo’fa method intervene,compared with the normal group,the expression of IL-1β in spinal dorsal horn and peripheral serum in the model group and the Bo’fa method group still increased obviously(P<0.05).That in the Bo’fa method group decreased obviously compared to the model group(P<0.05).[Conclusion]1.Bo’fa method can alleviat the hyperalgesia condition of CCI model rats,promote the regeneration of the injured sciatic nerve fibrous myelin and axons,reduce the edema of the cytoplasm and mitochondria in the Schwann cell.That provides the behavioral and morphological evidence of Bo’fa method treating the peripheral nerve injury diseases.2.Bo’fa method can increase the expression of the central inhibitory neurotransmitter 5-HT2A and GABA-B receptors in spinal cord of CCI model rats,participate in alleviating the derepression of the central system,restrain the transition of harmful sensory information,and then relieve the level of spontaneous pain caused by Neuropathic Pain effectively.3.Bo’fa method can decrease the expression of the proinflammatory cytokine IL-1β in spinal cord and the serum of CCI model rats,inhibit the central sensitization in the central system,and reduce the inflammatory sysmtoms of Neuropathic Pain in the peripheral system.That indicates that Bo’fa method has an important role of eliminating the inflammation in Neuropathic Pain effectively.4.The above conclusions confirm each other to indicate that Bo’fa method intervene can promote the hyperalgesia condition of Neuropathic Pain effectively,repair the injured nerve fibers,and relieve the level of abnormal pain. |
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