Objective: To explore the effect of electroacupuncture (EA) at "Shuigou"(GV6) and "Baihui"(GV20) on autophagy of hippocampal neurons in cerebral ischemia-reperfusion (I/R) injury rats.
Methods: Forty-eight healthy male SD rats were randomly divided into sham operation, model and EA groups, with 16 rats in each group. The rat model of cerebral I/R injury was established by occlusion of the middle cerebral artery (MCAO). Rats of the EA group received EA at GV26 and GV20 for 20 min, once daily for 5 days. The neurological function of rats in each group was evaluated by Longa neurological function score. The cerebral infarction volume was measured by TTC staining. The levels of IL-6, IL-18 and TNF-α in cerebrospinal fluid were detected by ELISA. Real-time PCR and Western blot were respectively used to detect the expressions of autophagy-related proteins AMPK, Beclin-1, VPS34 and LC3B.
Results: Compared with the sham operation group, neurological function scores of rats in the model group were significantly increased (P<0.01); the volume of cerebral infarction was significantly increased (P<0.01); the contents of IL-6, IL-18 and TNF-α in cerebrospinal fluid were increased (P<0.01, P<0.05); the mRNA expression levels of AMPK, Beclin-1, VPS34 and LC3B were significantly increased (P<0.01); the protein expressions of AMPK, Beclin-1, VPS34 and the ratio of LC3B-Ⅱ/LC3B-Ⅰ were increased (P<0.01, P<0.05). After intervention and in comparison with the model group, the neurological function scores were decreased (P<0.05); the cerebral infarct volume were decreased (P<0.05); the contents of IL-6, IL-18 and TNF-α in cerebrospinal fluid were decreased (P<0.05); the mRNA expressions of AMPK, Beclin-1, VPS34 and LC3B were significantly decreased (P<0.01); the protein expressions of AMPK, Beclin-1, VPS34 and the ratio of LC3B-Ⅱ/LC3B-Ⅰ were decreased (P<0.05, P<0.01).
Conclusion: EA can improve the neurological function and alleviate the degree of nerve injury in rats with cerebral I/R injury, which may be related to inhibiting the autophagy level of hippocampal neurons.
目的：探讨电针“水沟”“百会”穴对脑缺血再灌注（I/R）损伤大鼠海马神经元自噬的影响。方法：SD大鼠随机分为假手术组、模型组、电针组，每组16只。采用一侧大脑中动脉栓塞（MCAO）的方法复制I/R损伤大鼠模型。电针组予“水沟”“百会”电针治疗，20 min/次，1次/d，共5 d。以Longa神经功能评分法评价神经功能；TTC染色法测定大鼠脑梗死体积百分比；ELISA法检测脑脊液白细胞介素（IL）-6、IL-18、肿瘤坏死因子-α（TNF-α）含量；荧光定量PCR和Western blot法分别检测缺血区海马组织自噬相关蛋白腺苷酸活化蛋白激酶（AMPK）、Beclin-1、VPS34、LC3B mRNA和蛋白的表达。结果：与假手术组比较，模型组神经功能评分明显升高（P<0.01），大鼠脑梗死体积百分比明显增大（P<0.01），脑脊液IL-6、IL-18、TNF-α含量明显升高（P<0.01，P<0.05），海马组织AMPK、Beclin-1、VPS34蛋白和mRNA表达水平，以及LC3B mRNA表达水平和LC3B-Ⅱ/LC3B-Ⅰ比值均升高（P<0.01，P<0.05）。与模型组比较，电针组神经功能评分降低（P<0.05），脑梗死体积百分比缩小（P<0.05），脑脊液IL-6、IL-18、TNF-α含量降低（P<0.05），海马组织AMPK、Beclin-1、VPS34蛋白和mRNA表达水平，以及LC3B mRNA表达水平和LC3B-Ⅱ/LC3B-Ⅰ比值均降低（P<0.01，P<0.05）。结论：电针治疗可能通过抑制海马神经元自噬水平，缓解MCAO大鼠I/R损伤，减轻神经受损程度。.
Keywords: Autophagy; Cerebral ischemia-reperfusion injury; Electroacupuncture; Neuroinflammation.