Objective: To observe the effect of manual acupuncture stimulation of "Shuigou" (GV26) and "Neiguan" (PC6) on neurological function and expression of Caspase-3 and Caspase-9 in brain tissues around the intracerebral hematoma in rats with acute intracerebral hemorrhage (ICH), so as to explore its possible mechanisms underlying improvement of ICH.
Methods: Ninety-six male SD rats were randomly divided into 4 groups: control, model, acupoint and non-acupoint (24 rats in each group). The ICH model was established by injection of the rat's autologous blood into the caudate nucleus. According to the time-points of 6, 24, 48 and 72 h after ICH, each of the 4 groups was further divided into 4 subgroups. For rats of the acupoint group, the PC6 on both sides was manually stimulated by manipulating the needle with lifting-thrusting-twisting reducing techniques, while the GV26 was stimulated with strong "sparrow-pecking" method for 10 times, then, left the needles in the acupoints for 30 min. For rats of the non-acupoint group, two non-acupoints: mid-spot below the bilateral axilla and the spot 3 mm above the left side of the coccyx tip were stimulated with the same methods to PC6 and GV26, respectively. For rats of the 6 h and 24 h subgroups, the intervention was given once after waking up from modeling, and for those of the 48 and 72 h subgroups, the intervention was conducted once a day for 2 or 3 times, respectively. The neurological severity score (NSS) was used to evaluate the degree of neurological function. The immunoactivity (expression) of Caspase-3 and Caspase-9 proteins of the hematoma focus of the brain was detected by immunohistochemistry.
Results: Following modeling, the NSS and the expression levels of Caspase-3 and Caspase-9 proteins in the brain tissues surrounding the hematoma at each time-points (6, 24, 48 and 72h) after modeling were significantly increased in the model group relevant to the control group (P<0.01, P<0.05). Compared with the model group, the NSS at 72h and the expression levels of Caspase-3 and Caspase-9 proteins at 6, 24, 48 and 72h were significantly down-regulated in the acupoint group (P<0.05) rather than in the non-acupoint group (P>0.05).
Conclusion: Acupuncture of GV26 and PC6 can improve the neurological function in rats with ICH, which may be related to its function in reducing the expression of Caspase-3 and Caspase-9 proteins (apoptosis-related proteins) in the brain.
目的：观察大鼠脑出血急性期血肿周围脑组织中含半胱氨酸的天冬氨酸水解酶Caspase-3、Caspase-9表达随时间的动态变化及针刺干预对其影响，探讨针刺“水沟”“内关”对大鼠脑出血急性期神经细胞凋亡的影响及其可能的神经保护机制。方法：健康雄性大鼠随机分为对照组、模型组、穴位组、非穴组，每组24只。采用自体非肝素抗凝动脉血二次注入大鼠尾状核制作急性脑出血模型，符合纳入标准的大鼠再按出血后6、24、48、72 h分为4个时间点亚组。穴位组用提插捻转泻法刺激双侧“内关”，雀啄法强刺激“水沟”，两穴留针30 min；非穴组用提插捻转泻法刺激双侧腋中线下5 mm的2个非穴点，雀啄法强刺激尾骨尖左侧旁开3 mm的非穴点。6、24 h组于造模后大鼠清醒即刻干预1次，对应时间点取材；48、72 h组每日干预1次，对应时间点取材。采用神经损伤评分(NSS)法对大鼠行为学进行评分；用免疫组织化学法检测血肿周围脑组织Caspase-3、Caspase-9蛋白表达情况。结果：与同时点对照组比较，模型组大鼠出血后各时点NSS、血肿周围脑组织中Caspase-3及Caspase-9蛋白表达均明显升高(P<0.01，P<0.05)；与同时点模型组比较，穴位组72 h亚组NSS、各时点血肿周围脑组织中Caspase-3及Caspase-9蛋白表达均明显降低(P<0.05)；与同时点穴位组比较，非穴组72 h亚组NSS、各时点血肿周围脑组织Caspase-3及Caspase-9蛋白表达均明显升高(P<0.05)。结论：针刺“水沟”“内关”能改善出血后大鼠神经功能缺损体征，还可能通过降低出血后脑组织中Caspase-3、Caspase-9蛋白表达水平，抑制出血后由Caspase家族介导的神经元凋亡的发生，拮抗出血后脑组织损伤，保护神经元功能。.
Keywords: Acupuncture; Acute intracerebral hemorrhage; Cell apoptosis; Neiguan (PC6); Shuigou (GV26).