上海针灸杂志2017，Vol.36Issue(3)：343-353,11.DOI:10.13460/j.issn.1005-0957.2017.03.0343
芒针对促进大鼠急性脊髓损伤功能修复的机制研究
Mechanism Study of the Promoting Action of Elongated Needle Acupuncture on Functional Repair in Rats with Acute Spinal Cord Injury
摘要
Abstract
Objective To observe the effect of elongated needle acupuncture on inflammatory reactions and apoptosis in acute spinal cord injury, investigate if PI3K/Akt and MAPK/ERK signal transduction pathways are involved in the neuroprotective effect of elongated needle acupuncture and explore the promoting action of elongated needle acupuncture on spinal cord injury repair.Method One hundred and fifty adult male SD rats were randomized into groups A, B, C, D and E, 30 each. A model of moderate spinal cord injury was made by modified Allen's method. Group A received a sham operation without spinal cord injury and no elongated needle acupuncture. Group B did not receive elongated needle acupuncture after model making. Group C received elongated needle acupuncture after model making. Group D received an intrathecal injection of LY294002 at 0.5 hour before model making and elongated needle acupuncture after model making. Group E received an intrathecal injection of PD98059 at 0.5 hour before model making and elongated needle acupuncture after model making. Rat spontaneous activity was examined using the BBB rating. Inflammatory cytokines TNF-α, IL-6, IL-1β and NF-κB contents were measured by ELISA. The degree of apoptosis was determined by TUNEL. Bcl-2- and Bax-positive cell levels were measured by an immunohistochemical method. Spinal p-Akt and p-ERK expressions were determined by Western blot. In vivo expressions of Cyt-C and Caspase-3 were determined by RT-PCR. The in vivo inhibitory effect on downstream Akt and ERK signaling pathways was investigated using specific inhibitors LY294002 and PD98059.Result Inflammatory reactions and neuronal apoptosis due to the inhibition of PI3K/Akt and MAPK/ERK signal pathways were involved in the damage in a rat model of spinal cord injury. The neuroprotective effect of elongated needle acupuncture was related to a decrease in the number of Bax protein-positive neurons and an increase in the number of Bcl-2 protein-positive neurons. Elongated needle acupuncture treatment improved rat motor function, activated p-Akt and p-ERK in PI3K/Akt and MAPK/ERK signal pathways by down-regulating Bax protein expression and up-regulating Bcl-2 protein expression and inhibited the expression of Cyt-C, a key factor in the mitochondrial apoptosis pathway. TUNEL detected and inhibited the activation of caspase-3 cascade in neuronal apoptosis. Application of specific inhibitors LY294002 and PD98059 to PI3K/Akt and MAPK/ERK signaling pathways inhibited the expression p-Akt and p-ERK expressions.Conclusion JIN's three needle therapy plus mirror therapy for knee joint movement is an effective approach in treating hemiplegia after cerebral infarction, and it can improve the lower-limb motor function and the ADL.关键词
巨刺疗法/芒针/脊髓损伤/炎症反应/p-Akt/p-ERK/Bcl-2/Bax/Cyt-C/Caspase-3/凋亡/大鼠Key words
Big needle therapy/Elongated needles/Spinal cord injury/inflammatory reaction/P-Akt/P-ERK/Bcl-2/Bax/Cyt-C/Caspase-3/Apoptosis/Rats分类
医药卫生引用本文复制引用
李长明,谢尚举,全仁夫,王拓,杜伟斌,杨宗保..芒针对促进大鼠急性脊髓损伤功能修复的机制研究[J].上海针灸杂志,2017,36(3):343-353,11.基金项目
浙江省中医药科研计划项目(2008CB067) （2008CB067）
