| Objective:To observe the efficacy of the triple acupuncture and methylprednisolone combined therapy on SD rats with acute spinal cord injury, and to explore the potential mechanism of the triple acupuncture and methylpredinisolone combined therapy for acute spinal cord injury, providing experimental evidence for clinical treatment of acute spinal cord injury.Methods:We use improved clamped method to establish rat model of acute spinal cord injury.75rats with spinal cord injury were randomly divided into five groups,15rats in each group. Group A:the triple acupuncture therapy+methylprednisolone therapy, Group B:Du meridian acupoints+jiaji acupoints+body acupuncture and electroacupuncture of posterior limb+methylprednisolone therapy, Group C:methylprednisolone therapy, Group D:model group, Group E: sham-operated group. The Methylprednisolone therapy is began through intraperitoneal injection of methylprednisolone immediately after spinal cord injury; and in Electro-acupuncture treatment, rats in group A and B receive electro-acupuncture treatment4h after spinal cord injury, rats in group D and E receive no treatment. At day1,3and7, BBB(Basso Beattie Bresnahan) scores were calculated in each group to observe the neurofunction changes. The injured spinal cord tissue were observed for the changes of histological structure by HE staining, and the expression of CGRP as well as Caspase-3positive cell of rats after acute spinal cord injury in each group were observed by the way of immunohistochemistry.Results:1. Behavioral results:①Compared with group E, the BBB scores of rats from group A, B, C and D were significantly decreased at day1,3and7(P<0.05)②1day after treatment, there was no significant difference among the BBB scores of posterior limb in rats from group A, B, C and D (P<0.05);③3days after treatment, compared with group D, the scores of rats from group A, B and C were significantly increased (P<0.05);④7days after treatment, compared with group C and D, the scores of rats from group A and B were significantly increased (P<0.05), there were no significant difference between group A and B, compared with group D, the scores of rats from group C were significantly increased(P<0.05);⑤At day1,3and7after treatment, the scores in group A, B, C and D had improved at different degrees, with statistical significance (P<0.05);⑥the difference of the scores in group E were not statistically significant at various points in time (P>0.05)2. HE staining results:Varying degrees of hemorrhage, inflammatory cell infiltration, necrosis, cavity formation were seen in model group and each treatment group, with remnants of neuronal cell bodies become a sphere, karyopyknosis, edge-set and the disappearance of nucleus, as well as ill-defined boundary between white matter and gray matter. Compared with model group, pathological alteration of spinal cord tissue from group A, B and C were significantly reduced; At various points in time, neurons form from the sham-operated group have regular structural form, with compact white matter, no significant bleeding, edema, necrosis and inflammatory cell infiltration etc.3. Immunohistochemical results:①CGRP:positive cells were expressed in all groups, at1day and3day after spinal cord injury, the expression of positive cells in the rest groups were significantly lower than that in group E (P<0.05); At the seventh day after treatment, there was no significant difference in the expression of positive cells between group A、B and group E (P<0.05). The expression of positive cells in group C and D were significantly lower than that in group E (P<0.05); At various points in time, the expression of positive cells in group A and B was higher than that in group C and D (P<0.05); There was no significant difference between group A and B, and group C and D as well (P<0.05) The statistical differences which from1day and3day,1day and7day,3day and7day in group A and which from,1day and7day,3day and7day in group B were all obtained.②Caspase-3:at1,3and7day, the expression of positive cells in group A, B, C, and D were significantly higher than that in group E (P<0.05), and the expression of positive cells in group A, B and C were significantly lower than that in group D (P>0.05); At1day after treatment, there was no significant difference in the expression of positive cells among group A, B and C (P>0.05); At3and7day after treatment, the expression of positive cells in group A and B were lower than that in group C (P>0.05), however, there was no significant difference in the expression of positive cells between group A and B (P>0.05).There were statistical differences in the comparison at1day and3day,3day and7day in group A and B as well as in that at1day and3day,1day and7day,3day and7day in group C and D.Conclusions:1.The triple acupuncture therapy is effective in improving motor function of the posterior limb in SD rats with acute spinal cord injury.2The mechanism of the triple acupuncture therapy for SD rats with acute spinal cord injury may be related with improving the expression of CGRP, suppressing the expression of Caspase-3, thus inhibiting cell apoptosis, reducing secondary injury, protecting nerve cells and promoting regenerative repair of nerve cells.3The new way of selecting acupoints based on sporting point can be adopted in the clinical therapy. |
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