Wang Bo, Zhang Xiao-Ming, Wu Song, Huang Wei, Li Dan, Lu Wei, Xie Jun
Department of General Internal Medicine, Wuhan Seventh Hospital, Wuhan 430071, Hubei Province, China.
School of Acupuncture-Moxibustion, Orthopedics and Traumatology, Hubei University of CM, Wuhan 430061.
Zhongguo Zhen Jiu. 2019 Sep 12;39(9):957-62. doi: 10.13703/j.0255-2930.2019.09.011.
To explore the protective effect and apoptosis-related mechanism of electroacupuncture (EA) preconditioning in the rats with cerebral ischemia-reperfusion injury.
Sixty male SD rats, 3 months old, at SPF grade were randomized into a sham-operation group, an ischemia-reperfusion group and an EA preconditioning group, 20 rats in each one. In the ischemia-reperfusion group and EA preconditioning group, the modified MCAO suture-occlusion method was adopted to exert ischemia for 2 h and reperfusion for 3 h, and thus, the models of focal cerebral ischemia-reperfusion injury were prepared on the right side. In the sham-operation group, the right common carotid artery was separated and no more management was given. In the EA preconditioning group, EA at "Baihui" (GV 20), "Shenshu" (BL 23) and "Sanyinjiao" (SP 6) was provided before modeling, with disperse-dense wave, at 2 Hz/100 Hz, 1 mA in intensity. The stimulation for 15 min was taken as one unit (meaning electric stimulation for 10 min and needle retaining for 5 min without electric stimulation). Such preconditioning was repeated continuously for 4 times, totally for 1 h. The neuroethologic condition was assessed in 3 h of reperfusion in each group. TTC staining method was used to determine the percentage of cerebral infarction zone, TUNEL method was to determine the apoptosis index (AI) in hippocampal neuron and the immunohistochemical method (IHC) was to determine the protein expression of p53 and caspase-3.
Compared with the sham-operation group, the neuroethologic score, the percentage of cerebral infarction zone and neuronal AI were all increased obviously in the ischemia-reperfusion group (all <0.01). Compared with the ischemia-reperfusion group, the neuroethologic score, the percentage of cerebral infarction zone and neuronal AI were all reduced obviously in the EA preconditioning group (all <0.01). p53's nuclei and caspase-3's cytoplasms were stained. The positive cells of both of them were brown-yellow in color. In the sham-operation group, the structure of the right hippocampal CA3 neurons of rats was clear, with few positive cells. In the ischemia-perfusion group, the positive expressions of p53 and caspase-3 in the right hippocampal CA3 were increased obviously (<0.01). Compared with the ischemia-reperfusion group, the positive expressions of caspase-3 and p53 in the right hippocampal CA3 were significantly reduced in the EA preconditioning group (<0.01).
Electroacupuncture preconditioning relieves ischemic injury in brain tissue of rats probably through inhibiting the expressions of p53 and caspase-3 to resisting neuronal apoptosis.
探讨电针预处理对脑缺血再灌注损伤大鼠的保护作用及凋亡相关机制。
将60只3月龄SPF级雄性SD大鼠随机分为假手术组、缺血再灌注组和电针预处理组,每组20只。缺血再灌注组和电针预处理组采用改良线栓法阻断大脑中动脉2 h后再灌注3 h,制备右侧局灶性脑缺血再灌注损伤模型。假手术组仅分离右侧颈总动脉,不做进一步处理。电针预处理组于造模前在“百会”(GV 20)、“肾俞”(BL 23)和“三阴交”(SP 6)进行电针干预,采用疏密波,频率2 Hz/100 Hz,强度1 mA。每次刺激15 min为1个单元(即电针10 min,留针5 min不接电针),连续重复预处理4次,共1 h。各组于再灌注3 h时进行神经行为学评分。采用TTC染色法测定脑梗死面积百分比,TUNEL法测定海马神经元凋亡指数(AI),免疫组化法(IHC)测定p53和caspase-3蛋白表达。
与假手术组比较,缺血再灌注组神经行为学评分、脑梗死面积百分比及神经元AI均明显升高(均P<0.01)。与缺血再灌注组比较,电针预处理组神经行为学评分、脑梗死面积百分比及神经元AI均明显降低(均P<0.01)。p53在细胞核、caspase-3在细胞质染色,二者阳性细胞均呈棕黄色。假手术组大鼠右侧海马CA3区神经元结构清晰,阳性细胞较少。缺血再灌注组右侧海马CA3区p53和caspase-3阳性表达明显增加(P<0.01)。与缺血再灌注组比较,电针预处理组右侧海马CA3区caspase-3和p53阳性表达明显降低(P<0.01)。
电针预处理可能通过抑制p53和caspase-3表达,抵抗神经元凋亡,减轻大鼠脑组织缺血损伤。