| ObjectiveTo screen a stimulation method with the strongest protective effect on spinal cord tissue of acute spinal cord injury (ASCI) rats among the manual acupuncture, electroacupuncture (EA) and transcutaneous electrical acupoint stimulation (TAES), and to discuss neuron protective effects among three different point combinations under the same strongest stimulus on ASCI rats, and rehabilitation function and mechanism on chronic spinal cord injury (SCI) rats.MethodsWe applied modified Allen’s impact method (MASCIS Impactor, precision strike) to contribute the ASCI rat models. After the administration of three kinds of stimulation methods including manual acupuncture, EA and TAES, we used the BBB score at several time points to evaluate locomotion function changes of ASCI rats; HE staining and Nissl staining to observe the pathological morphology changes of injury spinal cord tissue; TUNEL staining to detect the apoptosis cells of injury spinal cord tissue; to analyze the effects of the three different stimulation methods.After EA administration with three different point combinations (Baihui and Fengfu group, Dazhui and Mingmen group, and Jiaji group) on ASCI rats, we used the BBB score to evaluate the locomotion function changes; HE staining to observe the morphological changes of the spinal cord; Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) staining to detect apoptosis cells of spinal cord tissue; and detected the change of malondialdehyde (MDA) level and superoxide dismutase (SOD) activity; to analyze effects of anti lipid peroxidation, pathological changes and cell apoptosis of EA with different point combinations in the acute phase after SCI.After applying EA with three different point combinations (Baihui and Fengfu group, Dazhui and Mingmen group, and Jiaji group) on ASCI rats, we used the BBB score to evaluate locomotion function changes of SCI rats; real-time quantitative PCR (RT-qPCR) detection and Western-blot to observe the expression of mRNA and protein of proinflammatory cytokines including interIeukin-6(IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α); to analyze the effects of EA intervention with different point combinations on inflammatory reaction and function recovery of SCI rats in acute phase. After applying EA with three different point combinations (Baihui and Fengfu group, Dazhui and Mingmen group, and Jiaji group) on chronic SCI rats, we used the BBB score to evaluate locomotion function changes of SCI rats; RT-qPCR detection and Western-blot to observe the mRNA and protein expression changes of brain derived neurotrophic factor (BDNF) and neurotrophin-3(NT3); to analyze the effects of EA intervention with different point combinations on inflammatory reaction and function recovery of SCI rats in chronic phase.ResultsAfter treatments of manual acupuncture, EA and TAES on ASCI rats, BBB score which was administrated at8.5hours of postsurgery, showed that locomotion function of rats was promoted in EA group compared with the model group (p<0.01). HE staining and Nissl staining showed that in sham group spinal cord and neurons displayed integrated infrastructures; but exudation, necrosis and edema were seen surrounding damaged tissue with irregular neurons morphology in model group; while the extents of neuronal damage in three treatment groups were between that in sham group and that in model group. TUNEL staining showed that the TUNEL-positive cells were widely distributed in both the white and gray matter, with the majority found in the white matter in model group, whereas, the quantities of TUNEL-positive cells in three treatment groups significantly decreased, especially in EA group.After treatments of EA with three different point combinations (Baihui and Fengfu group, Dazhui and Mingmen group, and Jiaji group) on ASCI rats, BBB score which was administrated at8.5hours of postsurgery, showed that locomotion functions of rats in three EA groups were improved compared with the model group (p<0.01), whereas, Jiaji group had the most significantly improvement (p<0.01); HE staining showed that in sham group spinal cord displayed integrated infrastructures, while hemorrhage and necrosis were seen surrounding damaged tissue in model group; and the extents of hemorrhage and necrosis were mild compared with the model group. TUNEL staining displayed a large number of TUNEL-positive cells in model group, while apoptosis cells in three EA groups were decreased, especially in Jiaji group. Meanwhile, the SOD activity increased, and MDA levels decreased in the three EA groups compared with the model group (p<0.05), with no difference among three EA groups (p>0.05).After treatments of EA with three different point combinations (Baihui and Fengfu group, Dazhui and Mingmen group, and Jiaji group) on SCI rats in acute phase, BBB score which was administrated at8.5hours of postsurgery, showed that locomotion function of rats in three group were promoted compared with the model group (p<0.05), whereas, Jiaji group had significant improvement (p<0.01). QRT-PCR and Western-blot showed that in model group the mRNA and protein expression of TNF-a, IL-1β and IL-6significantly increased compared with sham group (p<0.01), while that in all three treatment groups were decreased compared with the model group (p<0.01), whereas, Jiaji group decreased most significantly (p<0.05).After treatments of EA with three different point combinations on SCI rats in chronic phase, BBB score at each time point (1st day,1week,3week,7week postsurgey) showed gradually increase of locomotion function in each treatment group compared with the model group (p<0.01), but the most significantly increase in Jiaji group (p<0.05). QRT-PCR and Western-blot showed that in model group the mRNA and protein expression of BDNF and NT-3significantly increased after7weeks postsurgery compared with model group (p<0.01), whereas, Jiaji group increased most significantly (p<0.01).Conclusion1. Manual acupuncture, EA and TAES alleviated the acute injury of rats induced by SCI, protected the spinal cord tissue, and reduced cells apoptosis after SCI, especially EA had the most significant effect, and promoted early locomotion function.2. EA administrations with different point combinations on ASCI rats had the neuroprotection functions on anti lipid peroxidation, anti-inflammatory and reduction of cells apoptosis, and locomotion function recovery in the acute phase after SCI, whereas, Jiaji group had the most remarkable anti-inflammatory function.3. EA administrations with different point combinations on SCI rats in chronic phase, greatly promoted neuronal recovery and function rehabilitation by increasing the expression of BDNF and NT-3in local injury spinal cord, whereas, the Jiaji group had the most remarkable effect. |
PDF Full Text Request