| [Objective]Through the behavior research of SFI and thermal pain tolerance domain and the morphological study on the two parts of spinal cord and the injury point of sciatic nerve to confirm that EA’s treatment to SNI is effective.Through the expression of NGF,MBP in the spinal cord and the injury point of sciatic nerve and the expression of MBP and MBP antibody in serum, from myelin repair point to view further explanation of the mechanism of EA’s treatment to SNI.[Method]Choosing the sciatic nerve clipping injury model of SD rats as the exper-imental animals;Animals were divided into5groups:normal group sham operation group,model group,model control group, EA group. EA group treated by electric acupuncture apparatus. Current was2mA. Frequency was5HZ. The wave choosed dilate-tional wave. The points we selected were Huan tiao, Yin men, Yang lingquan and Cheng shan in the injured side.Behavior used heat pain tolerance threshold to observe the recovery of the rats’sensory function;Used SFI detection to observe the recovery of the rats’ motor function.The morphology used HE staining and myelin staining (Solochrome Cyanine me-thod). HE staining and myelin staining of the spinal cord and the injury of SN were mainly used to observe the injury and recovery of neurons, nerve fibers, myelin sheath; HE staining of gastrocnemius and myocyte diameter measurement were used to observe the injury and recovery of target tissues; The dyed film observed and photographed by the400×light microscope.Histological used immunohistochemical method to qualitative analysis the expression of NGF and MBP in the spinal cord and the injury point of SN;The dyed film observed and photographed by the400x light microscope; Used Image Proplus6.0software to analysis and calculate the photos’ average optical density.Molecular biology used ELISA method to quantitative measure the expression of MBP and MBP antibody in the serum;Using Microplate reader to test optical density of each hole; Using EXCEL to make the standard curve and calculate the concentration of each sample.Statistics used SPSS Statistics17.0software to analysis datas, datas were finally expressed as mean±standard deviation (χ±s).[Result]1Behavioral results1.1The results of thermal pain tolerance7days after modeled, the right (ipsilateral) of model group increased obviously compared with normal group (P<0.05), sham operation group showed no significant differen cecompared with normal group (P>0.05).20times after Intervented, right (ipsilateral) of model group and model control group were significantly higher than that of normal group (P<0.05), EA group were decreased significantly compared with the model group and the model control group (P<0.05), EA group were no significant difference with normal group (P>0.05).1.2The results of SFI7days after modeled, model group were significantly lower than that of normal group (P<0.05).20times after intervented, EA group, model group and model control group were significantly lower than normal group(P<0.05), there were no significantly difference between the EA group and model group, model control group(P>0.05).2The morphological results2.1The results of gastrocnemius HE staining7days after modeled, normal group and sham operation group:muscle cells distributed in the musclefibers’surrounding, structureswereclear, muscle cells arranged in neat rows, with uniform diameter, muscle fibers’ gaps were moderate; model group:some muscle nuclei s appeared within shift, structures were fuzzy, muscle cells were arranged in order,diameters were different, the gaps were smaller, muscle fibernecros is. Myocyte diameter of mode lgroup were significantly smaller than normal group (P<0.05), no significant difference between normal group and sham operation group (P>0.05).20times after intervented, model group and model control group:the nucleus of muscle cells were divorced from muscle cells, thier arrangement were very confusion. Muscle fibers were necrosis, the gap between the muscle fibers were bigger. The nerve fibers in Muscle fibers appeared disintegration. EA group: the nucleus of muscle cells’ structure were clear, they distributed in the surrounding of muscle fibers. Muscle cells arranged in neat rows, their size were uniform, the gap between muscle fibers were slightly big. The muscle cells’ diameter of model group and model control group were significantly smaller than normal group (P<0.05), The muscle cells’ diameter of EA group were bigger than model group and model control group (P<0.05),but were smaller than normal group (P>0.05).2.2The results of sciatic nerve HE staining7days after modeled,normal group and sham operation group:the nerve axon and myelin were integrity, arranged closely. Schwann cells wrapped the edge of myelin, their nucleus were visible. The shape of capillary lumen between nerve fibers were rules. Model group:sciatic nerve appeared demyelination, part of axonals were disintegration. Schwann cells apoptosised, their nuclears were fuzzy, The shape of capillary lumen between nerve fibers were irregular.20times after intervention,model group and model control group:nerve fibers were swelling and scattered. sciatic nerve appeared demyelination, axonals were disintegration.Myelin and axonal mixed to form ovale. Schwann cells appeared proliferation;EA group:to some extent the lesions were repaired. In the surrounding of sciatic nerve we could see regenerated nerve fibers. And there were thick myelin wrapping regenerated nerve, their axonals were clear.There were a lot of Schwann cells proliferation.2.3The results of spinal cord HE staining7days after modeled,normal group and sham operation group:the nucleis of neuronal were clear. Nissl body were big and clear. Model group:neuronal lossed, nucleolus were blurred,Nissl body decreased.20times after intervented, model group and model control group:neurons were significantly decreased, swelled and cracked severity. A small amount of glial cell proliferated. EA group:neurons and nuclears’structure were clear, neurons swelled a lesser extent. There were a small amount of new neurons.2.4The results of the sciatic nerve myelin staining7days after modeled, normal group and sham operation group:myelin were clear, arounded the axons closely, arranged neatly.Model group:most of nerve myelins were collapse, degeneration, delamination and fusion. Axons were atrophy. A few myelin didn’t degenerate, but the thickness of these mylin were thinner. Schwann cells appeared apoptosis.20times after intervented,model group and model control group:the degree of myelination was very low. The vast majority of myelins were degeneration, delamination and fusion. The axons were destruction. The gaps between nerve bund-les were larger. The arrangement of nerve bundles were disorder. Nerves damaged badly. EA group:the degree of myelination was very high.Myelin were clear, arounded the axons closely, arranged neatly. In the periphery of myelin, we conuld saw hyperplasia Schwann cells. The nucleus of Schwann cells were clear. There were still small part of myelins were collapse, degeneration and delamination.2.5The results of spinal cord myelin staining7days after modeled,normal group, sham operation group:the structuralof myelins in the white matter of spinal cord were integrity,tight and their arrangement were neat. The axons were clear. The nucleus of the peripheral glial cells were clear. Model group:myelins were flaky myelinolysis, fractureand structural were disorder. The axons were swelling. The glial cells began to apoptosis.There were many vacuoles in the field of vision.20times after intervented,model group and model control group:The myelins in the white matter of spinal cord were loose. Myelindegree was very low. The swelling degree of axons were aggravate. The axons were disintegration, formed capsular space. There were a small amount of apoptosis glial cells in the field of vision. The nucleus were fuzzy. EA group:Myelindegree was very high. The myelin and axons’structure were complete, but the myelins were thin. The diameter of axons were small. A large number of glial cells proliferated in field of vision. The nucleus were clear.3The results of ELISA3.1Changes of MBP content in serum7days after modeled, the concentration of MBP in serum of model group were hign than those in normal group (P<0.05). Sham operation group had no obvious difference with normal group (P>0.05).20times after intervented, the concentration of MBP in serum of model group and model control group were higher than those in normal group (P<0.05). EA group were significantly lower than those of model group and model control group (P<0.05). The expression of MBP in EA group were near to normal group, but were slightly higher than normal group.3.2Changes of MBP antibody content in serum7days after modeled, the concentration of MBP antibody in serum of the model group were higher than those in the normal group (P<0.05). Sham operation group had no obvious difference with normal group (P>0.05).20times after intervented, the concentration of MBP antibody in serum of model group and model control group were higher than thoes in the normal group (P<0.05). EA group were significantly lower than thoes in model group and model control group (P<0.05). The expression of MBP antibody in EA group were near to normal group, but were slightly higher than normal group.4The results of immunohistochemistry4.1The expression of MBP in the spinal cord and injury point of sciatic nerve7days after modeled, the average optical density of MBP’s concentration in the sciatic nerve and spinal cord of modle group were significantly higher than thoes in normal group (P<0.05). Sham operation group had no obvious difference with the normal group (P>0.05).20times after intervented, the average optical density of MBP’s concentrate-ion in the sciatic nerve and spinal cord of modle group were significantly higher than thoes in normal group (P<0.05).Model control group had no significant difference with model group(P>0.05). EA group were markedly lower than thoes of model group and model control group (P<0.05). The expression of MBP in EA group were near to normal group, but were slightly higher than normal group.3.2The expression of NGF in the spinal cord and injury point of sciatic nerve7days after modeled, the average optical density of NGF’s concentration in the sciatic nerve and spinal cord of modle group were significantly higher than thoes in normal group (P<0.05). Sham operation group had no obvious difference with the normal group (P>0.05).20times after intervented, the average optical density of NGF’s concentrate-ion in the sciatic nerve and spinal cord of modle group were higher than thoes in normal group (P<0.05).Model control group had no significant difference with model group (P>0.05). EA group were markedly higher than thoes of model group and model control group (P<0.05). The expression of NGF in EA group were higher than normal group.[Conclusion]1EA can restore the rat’s response to pain effectively, promote the recovery of the rat’s sensory function of sciatic nerve. EA can prevent muscle atrophy effectively, so as to promote the recovery of motor function of rat.2EA can protect rat’s neurons,promote the proliferation of glial cells, reduce the myelin and axon’s disintegration, promote the myelin and axon’s regeneration, so as to promote the repair of injury of sciatic nerve.3EA can promote the expression of NGF in spinal cord and injury point of sciatic nerve of the rat,accelerate the rat’s nerve ability to regenerate.4EA can decrease the expression of MBP in spinal cord, injury point of sciatic nerve and serum of the rat, thereby reduce the myelin’s loss, promote the myelin’s repair.5EA can decrease the expression of MBP antibody in serum of rat, reduce rat’s autoimmune reaction. Thereby,EA can reduce the second damage to the myelin and protect myelin.In conclusion, the results of this study confirme that EA is suitable for the sciatic nerve injury, can promote the expression of NGF, reduce the expression of MBP and MBP antibodies, can accelerate the process of the myelin’s repair then speed up the process of the sciatic nerve injury’s repair. |
PDF Full Text Request