针刺对脑缺血再灌注损伤模型大鼠海马细胞凋亡的影响

The effects of acupuncture on apoptosis of hippocampal cells in rats with cerebral ischemia-reperfusion injury model

目的:探讨针刺对脑缺血再灌注损伤(CIRI)大鼠海马区细胞凋亡的影响及机制。方法:SD大鼠共75只,随机分为假手术组(sham)、脑缺血再灌注损伤模型组(CIRI)和针刺治疗组(CIRI+AC),利用大脑中动脉栓塞法制备CIRI大鼠模型,CIRI+AC组大鼠接受针刺“大椎”、“水沟”、“百会”穴治疗。Garcia评分法检测大鼠神经功能,TTC染色法检测大鼠脑梗死面积,免疫组织化学染色检测海马区caspase-3阳性细胞数量,Western Blot检测海马区caspase-9表达,TUNEL检测细胞凋亡。结果:针刺前,CIRI组和CIRI+AC组神经功能评分较sham组显著降低(P<0.01);针刺24、72 h后,CIRI+AC组神经功能评分较CIRI组显著升高(P<0.01),脑梗死面积比缩小(P<0.05),患侧脑组织细胞凋亡率降低(P<0.01);与针刺24 h后比较,72 h后CIRI组患侧脑组织细胞凋亡率及海马区caspase-3阳性细胞数量增加(P<0.05),CIRI+AC组患侧脑组织细胞凋亡率及海马区caspase-3阳性细胞数量明显降低(P<0.05)。与CIRI组比较,针刺24 h后CIRI+AC组患侧海马区caspase-9表达显著升高(P<0.01),针刺72 h后caspase-9表达显著降低(P<0.01)。结论:持续针刺可改善大鼠神经功能和细胞凋亡情况,且针刺对CIRI大鼠海马区细胞凋亡的抑制具有时效性。

Objective:To explore the effects and mechanism of acupuncture on apoptosis in the hippocampus of rats with cerebral ischemia-reperfusion injury(CIRI).Methods:Seventy-five SD rats were randomly divided into sham group,CIRI group and CIRI+AC group,the CIRI rat model was prepared by middle cerebral artery occlusion.The rats in the CIRI+AC group received acupuncture at Dazhui,Shuigou and Baihui.All rats were evaluated with Garcia neurological function score.TTC staining detects the cerebral infarction area in rats.Immunohistochemical staining detects the number of caspase-3 positive cells in the hippocampus.The expression of caspase-9 was detected by Western Blot,and the apoptosis rate was detected by TUNEL.Results:Before acupuncture,neural function scores in the CIRI group was lower than those of the sham group(P<0.01),and after acupuncture for 24 and 72 h,the neurological function scores of the CIRI+AC group were significantly higher than those of the CIRI group(P<0.01),while the area ratio of cerebral infarction was decreased(P<0.05),the apoptosis rate of ischemia brain tissue was decreased(P<0.01).After 72 hours,the apoptosis rate of ischemia brain tissue was increased(P<0.05),while the number of caspase-3 positive cells in the CIRI+AC group was decreased(P<0.05).Compared with the CIRI Group,the expression of caspase-9 in the hippocampus of CIRI+AC group increased significantly at 24 h after acupuncture(P<0.01),and decreased significantly at 72 h after acupuncture(P 0.01).Conclusion:Acupuncture can improve the neurological function and reduce the apoptosis in CIRI rats,and the inhibition of apoptosis in the hippocampal region of CIRI rats by acupuncture was time-sensitive.