Effects of Electroacupuncture at Governor Vessel Acupoints on Neurotrophin-3 in Rats with Experimental Spinal Cord Injury.

In an effort to explore new, noninvasive treatment options for spinal cord injuries (SCI), this study investigated the effects of electroacupuncture (EA) for SCI rat models. SCI was induced by a modified Allen's weight-drop method. We investigated the response of EA at Dazhui (GV 14) and Mingmen (GV 4) acupoints to understand the effects and mechanisms of EA in neuroprotection and neuronal function recovery after SCI. BBB testing was used to detect motor function of rats' hind limbs among groups, and EA was shown to promote the recovery of SCI rats' motor function. Nissl staining showed a restored neural morphology and an increase in the quantity of neurons after EA. Also, the antiapoptosis role was exposed by TUNEL staining. Western blotting analysis was used to determine the protein expression of neurotrophin-3 (NT-3) in spinal cord tissue. Compared to the sham group, the expression levels of NT-3 were significantly decreased and EA was shown to upregulate the expression of NT-3. The present study suggests that the role of EA in neuroprotection and dorsal neuronal function recovery after SCI in rats, especially EA stimulation at GV 14 and GV 4, can greatly promote neuronal function recovery, which may result from upregulating the expression of NT-3.

Electroacupuncture at Dazhui (GV14) and Mingmen (GV4) protects against spinal cord injury: the role of the Wnt/ß-catenin signaling pathway.

Electroacupuncture at Dazhui (GV14) and Mingmen (GV4) on the Governor Vessel has been shown to exhibit curative effects on spinal cord injury; however, the underlying mechanism remains poorly understood. In this study, we established rat models of spinal cord injury using a modified Allen's weight-drop method. Ninety-nine male Sprague-Dawley rats were randomly divided into three equal groups: sham (only laminectomy), SCI (induction of spinal cord injury at T10), and EA (induction of spinal cord injury at T10 and electroacupuncture intervention at GV14 and GV4 for 20 minutes once a day). Rats in the SCI and EA groups were further randomly divided into the following subgroups: 1-day (n = 11), 7-day (n = 11), and 14-day (n = 11). At 1, 7, and 14 days after electroacupuncture treatment, the Basso, Beattie and Bresnahan locomotor rating scale showed obvious improvement in rat hind limb locomotor function, hematoxylin-eosin staining showed that the histological change of injured spinal cord tissue was obviously alleviated, and immunohistochemistry and western blot analysis showed that Wnt1, Wnt3a, ß-catenin immunoreactivity and protein expression in the injured spinal cord tissue were greatly increased compared with the sham and SCI groups. These findings suggest that electroacupuncture at GV14 and GV4 upregulates Wnt1, Wnt3a, and ß-catenin expression in the Wnt/ß-catenin signaling pathway, exhibiting neuroprotective effects against spinal cord injury.

[Proteome analysis on the mechanism of electroacupuncture in relieving acute spinal cord injury at different time courses in rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the differentially expressed proteins in the spinal cord at different time courses after acute spinal cord injury (ASCI) in the rat, so as to study its underlying mechanism in im-proving spinal traumatic injury. METHODS: A total of 105 male SD rats were randomized into normal control, model-6 h, EA-6 h, model-24 h, EA-24 h, model-48 h, EA-48 h groups, with 15 cases in each. ASCI model was established by using modified Allen's method. EA (2 Hz, 2-5 mA) was applied to "Mingmen" (GV 4) and "Dazhui" (GV 14) for 30 min. The injured spinal cord tissue (T10 -T11) was collected 6 h, 24 hand 48 h after ASCI and EA treatment, weighted and stored under -80 degrees D till detection. Two-dimensional gel electrophoresis (2-DE) was used to separate total proteins of the spinal tissue, followed by protein extraction and quantitation, 2-D gel image analysis, matrix assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS), and databases-searching for identification of the differentially-expressed proteins. RESULTS: A total of 10 differentially expressed proteins were identified in the present study. At 6 h, compared with control group, of the 5 types of spinal differential proteins, 4 were upregulated in the expression after ASCI, and the rest one was downregulated; while after EA, ASCI-induced expression changes in 4 of the 5 differential proteins were reversed. At 24 h after ASCI, 7 types of differential proteins were identified. Compared with control group, 6 differential proteins were upregulated, and the rest one was downregulated in model group. Compared with model group, ASCI-induced expression changes in 6 of the 7 differential proteins were reversed by EA. At 48 h after ASCI, a total of 8 types of differential proteins were identified. Compared with control group, 6 differential proteins were upregulated in the expression, and the rest two downregulated in model group. Compared with model group, ASCI-induced expression changes in 5 of the 8 differential proteins were reversed by EA. Along with the increased time and treatment, 24 h vs 6 h, two more differential proteins were identified, i.e., nucleoside diphosphate kinase and triosephosphate isomerase 1 (TPI 1). 48 h vs 24 h, 3 more differential proteins were identified, i.e., dihydrolipoamide dehydrogenase, malate dehydrogenase 1, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH); but two proteins disappeared, i.e., nucleoside diphosphate kinase, and ubiquitin-conjugating enzyme E2N. The identified differential proteins involving the effects of EA in regulating cellular energy metabolism, DNA repair, cellular generation, differentiation, apoptosis, etc. CONCLUSION: Proteome analysis indicates that in ASCI rats, some differentially expressed proteins involving energy metabolism, neuronal apoptosis reduction, protein-degradation inhibition may contribute to the effect of EA in repairing the traumatic spinal tissue.