Yang Cheng, Tan Cheng-Fang, Yang Zhu-Xin, Huang Si-Qin
College of Traditional Chinese Medicine, Chongqing Medical University, Chongqing 400016, China.
Department of Preventive Medicine, Banan District Hospital of Traditional Chinese Medicine, Chongqing 401320.
Zhen Ci Yan Jiu. 2021 Dec 25;46(12):987-95. doi: 10.13702/j.1000-0607.201117.
To explore the effect of electroacupuncture (EA) combined with Schwann cell (SC) transplantation (SCT) on remyelination of axons and neuregulin (Nrg1) in rats with compressed spinal cord injury(CSCI),so as to explore the mechanism of EA and SCT underlying improvement of CSCI.
SD female rats were randomly divided into normal, mo-del, EA, SCT, and EA+ SCT groups (=40 per group). A self-developed model of spinal compressed injury was adopted in this study. Rats of the model group were administrated laminectomy without treatment. Rats in the EA group were administrated EA stimulation at "Dazhui"(GV14), "Mingmen"(GV7), bilateral "Zusanli" (ST36) and "Taixi" (KI3) on the second day post-surgery for 10 min. Rats in the SC group were administrated SCT at 1 week post-surgery, and in the EA+SC group were given EA stimulation combined with SCT. The injured spinal cord tissue was obtained 0, 2, 4 and 8 weeks after compressed spinal injury. The functional recovery was assessed by Basso-Beattie-Bresnahan (BBB) score. The survivals and migration of SC after transplantation, myelination were observed by immunofluorescence. The ultrastructure of myelin in injured site was observed by transmission electron microscope，and the expression levels of glial fibrillary acidic protein (GFAP), protein zero(P0), and Nrg1 and Nrg1-ntf (cleavage protein of Nrg1) proteins of the spinal cord were detected by Western blot.
Compared with the normal group, BBB scores in the model group was significantly decreased（<0.05）,nervous fibers were demyelinated, numbers of normal and newborn myelination were decreased(<0.05)，expression of P0 was significantly increased (<0.05)，expression of GFAP was significantly increased(<0.05)，and the expression levels of Nrg1 and Nrg1-ntf proteins were decreased(<0.05). In comparison with the model group, the BBB scores in the EA, SCT and EA+SCT groups were significantly increased（<0.05,<0.01）, demyelination was improved, numbers of normal and newborn myelinations were increased(<0.05,<0.01)，expressions of P0 were significantly increased (<0.05,<0.01)，expressions of GFAP were significantly decreased(<0.05)，and the expression levels of Nrg1 and Nrg1-ntf proteins were increased (<0.05, <0.01).The differences were most significant in the EA+SCT group among the three groups.
EA can improve the locomotor function in CSCI rats, which may be rela-ted to its functions in promoting the survival and migration of transplanted SC and remyelination, and increasing the expressions of Nrg1 and its cleavage protein after SC transplantation.
探讨电针（EA）联合雪旺细胞（SC）移植（SCT）对脊髓压迫性损伤（CSCI）大鼠轴突髓鞘再生及神经调节蛋白（Nrg1）的影响，以探究EA和SCT改善CSCI的作用机制。
将雌性SD大鼠随机分为正常组、模型组、EA组、SCT组和EA+SCT组（每组n=40）。本研究采用自行建立的脊髓压迫损伤模型。模型组大鼠行椎板切除术但不做处理。EA组大鼠于术后第2天在“大椎”（GV14）、“命门”（GV7）、双侧“足三里”（ST36）和“太溪”（KI3）进行电针刺激10分钟。SC组大鼠于术后1周进行SC移植，EA+SC组大鼠给予电针刺激并联合SC移植。在脊髓压迫损伤后0、2、4和8周获取损伤脊髓组织。通过Basso-Beattie-Bresnahan（BBB）评分评估功能恢复情况。通过免疫荧光观察移植后SC的存活和迁移、髓鞘形成情况。用透射电子显微镜观察损伤部位髓鞘的超微结构，并用蛋白质免疫印迹法检测脊髓中胶质纤维酸性蛋白（GFAP）、蛋白零（P0）以及Nrg1和Nrg1-ntf（Nrg1的裂解蛋白）的表达水平。
与正常组相比，模型组BBB评分显著降低（P<0.05），神经纤维脱髓鞘，正常和新生髓鞘数量减少（P<0.05），P0表达显著增加（P<0.05），GFAP表达显著增加（P<0.05），Nrg1和Nrg1-ntf蛋白表达水平降低（P<0.05）。与模型组相比，EA组、SCT组和EA+SCT组BBB评分显著升高（P<0.05，P<0.01），脱髓鞘情况改善，正常和新生髓鞘数量增加（P<0.05，P<0.01），P0表达显著增加（P<0.05，P<0.01），GFAP表达显著降低（P<0.05），Nrg1和Nrg1-ntf蛋白表达水平升高（P<0.05）。三组中EA+SCT组差异最为显著。
EA可改善CSCI大鼠的运动功能，这可能与其促进移植SC的存活和迁移、髓鞘再生以及增加SC移植后Nrg1及其裂解蛋白的表达有关。
