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目的:观察针刺联合亚低温对脑缺血再灌注损伤(CIRI)大鼠脑组织miRNA-204及其靶基因表达的影响,探讨针刺联合亚低温对CIRI的保护机制.方法:将60只Sprague-Dawley大鼠按随机数字表法分为空白组、假手术组、模型组、针刺组、亚低温组和针刺联合亚低温组,每组10只.除空白组和假手术组外,其余4组大鼠均制备CIRI模型.造模成功后,空白组常规饲养72 h,不做任何处理;假手术组与模型组常规饲养72 h,术后只捆绑,不予其他处理;针刺组常规饲养72 h,捆绑后予以针刺大椎、百会和水沟;亚低温组常规饲养72 h,捆绑后用亚低温法维持大鼠温度72 h;针刺联合亚低温组常规饲养72 h,捆绑后予以亚低温及针刺干预,针刺方法同针刺组,亚低温方法同亚低温组.干预72 h后,观察各组大鼠神经功能缺损评分、脑梗死面积比值、缺血侧海马组织miRNA-204及其靶基因Map3k8、Ntrk2和Ppp3r1的表达.结果:干预前,与空白组和假手术组相比,造模大鼠的神经功能缺损评分及梗死面积均明显增加,组间差异均具有统计学意义(均P<0.01),说明造模成功.干预后,与模型组相比,3个干预组神经功能缺损评分均显著降低(均P<0.01);与针刺组和亚低温组比较,针刺联合亚低温组梗死面积比值显著减少(均P<0.01);在miRNA-204表达方面,与模型组相比,3个干预组大鼠的脑组织中miRNA-204表达均显著降低(均P<0.05),其中针刺联合亚低温组下调miRNA-204表达的趋势最显著(P<0.01);与针刺组和亚低温组相比,针刺联合亚低温组Map3k8表达显著升高(均P0.05).结论:针刺、亚低温、针刺联合亚低温干预均可降低CIRI大鼠的神经功能缺损评分,缩小脑梗死面积,其中针刺联合亚低温作用最为显著;在调节miRNA-204靶基因Map3k8表达方面,针刺联合亚低温的作用优于单独针刺或亚低温干预,但在调节Ntrk2和Ppp3r1表达方面,针刺组、亚低温组和针刺联合亚低温组作用相当.“,”Objective: To explore the protective mechanism of acupuncture plus mild hypothermia for cerebral ischemia-reperfusion injury (CIRI) by observing the effects of acupuncture plus mild hypothermia on miRNA-204 and its target gene expressions in CIRI rat brain tissues. Methods: Sixty Sprague-Dawley rats were divided into a blank control group, a sham operation group, a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group according to the random number table method (n=10). Except for the blank control group and the sham operation group, rats in the other 4 groups received CIRI modeling. After the model was successfully established, rats in the blank control group were bred routinely for 72 h without any interventions; rats in the sham operation group and the model group were bred routinely for 72 h, and only received binding without other interventions after surgery; rats in the acupuncture group were bred routinely for 72 h, and received acupuncture at Dazhui (GV 14), Baihui (GV 20) and Shuigou (GV 26) after binding; rats in the mild hypothermia group were bred routinely for 72 h, and received mild hypothermia intervention for 72 h after binding; rats in the acupuncture plus mild hypothermia group were bred routinely for 72 h, followed by receiving acupuncture as in the acupuncture group and mild hypothermia therapy as in the mild hypothermia group after binding. The neurological impairment score, cerebral infarction area ratio, the expressions of miRNA-204 and its target genes including Map3k8, Ntrk2 and Ppp3r1 in the ischemic hippocampus of each group were observed after 72 h of intervention. Results: Before intervention, compared with the blank control group and the sham operation group, the neurological impairment scores and the infarction area ratios of the modelled rats were statistically significantly increased (all P<0.01), indicating that the model was successful. After intervention, compared with the model group, the neurological impairment scores of the three intervention groups were significantly reduced (all P<0.01); compared with the acupuncture group and the mild hypothermia group, the infarction area ratio in the acupuncture plus mild hypothermia group was significantly reduced (both P<0.01); compared with the model group, the three intervention groups showed significant inhibition of miRNA-204 expression in brain tissues (all P<0.05), which was most significant in the acupuncture plus mild hypothermia group (P<0.01); compared with the acupuncture group and the mild hypothermia group, the Map3k8 expression in the acupuncture plus mild hypothermia group was significantly increased (both P0.05). Conclusion: Acupuncture, mild hypothermia, and acupuncture plus mild hypothermia reduced the neurological impairment score and the cerebral infarction area in CIRI rats, while acupuncture plus mild hypothermia showed the most significant effect. In regulating miRNA-204 target gene expressions, acupuncture plus mild hypothermia showed the same effect on Ntrk2 and Ppp3r1 expressions, while better effect on Map3k8 expression compared with either acupuncture or hypothermia.