目的 观察电针长强穴对FMR1基因敲除小鼠不同脑区CREB及p-CREB蛋白表达量影响，以探讨针刺的多脑区联动效应。方法 选取适龄FMR1基因敲除小鼠，通过PCR鉴定其基因表型。采用随机数字表法分为空白组、非经非穴组和长强组，每组8只，共24只。空白组在相同条件下仅给予抓取动作；非经非穴组取选取肋弓最低点上1cm处；长强组取长强穴；2组针刺组采用电针仪进行干预,电针刺激频率2 Hz，强度2mA，连续波，每日20 min，连续干预14 d。干预结束后通过免疫组化法检测海马、皮质及小脑CREB及p-CREB蛋白的表达情况。结果 ①空白组海马的CREB表达量较皮质（P<0.05）、小脑（P<0.001）显著升高；非经非穴组及长强组皮质的CREB表达量较海马（P<0.05，P<0.01））、小脑（P<0.05）显著升高；②空白组小脑的p-CREB及p-CREB率的表达较皮质（P<0.01，P<0.001））、海马（P<0.05，P<0.001）显著升高；非经非穴组的小脑p-CREB表达量较海马（P<0.01）显著升高；非经非穴组小脑p-CREB率较皮质（P<0.01）、海马（P<0.05）具有显著升高；二者在长强组中三个脑区均无显著性差异（P＞0.05）。轮廓分析结果示：①CREB平行轮廓分析示P＜0.05，提示三个脑区的总体轮廓不平行，即脑区的联动变化不一致。②p-CREB及p-CREB率水平轮廓分析示P＜0.01，提示三个脑区联动变化一致，且各组间变化程度一致，但各组中蛋白表达不相等，其中小脑最高。 结论 基于轮廓分析能够初步观察针刺多脑区联动效应，且针刺刺激可能通过影响CREB表达使得FMR1基因敲除小鼠多脑区效应方式发生改变。
Objective This study was to investigate the effect of electroacupuncture at GV1 acupoint on the expression of CREB and p-CREB protein in different brain regions of FMR1 knockout mice, in order to explore the multi-brain linkage effect of acupuncture.Methods The FMR1 knockout mice of the appropriate age were selected and their gene phenotypes were identified by PCR. The random number table method was divided into blank group, non-acupoint group and GV1-acupoint group, with 8 rats in each group, a total of 24 rats.The blank group was given only the grasping action under the same conditions.The non-acupoint group was taken 1 cm upward from the lowest point of the rib arch.The GV1-acupoint group took GV1-acupoint group. The two groups of acupuncture groups were treated with an electroacupuncture instrument. The electroacupuncture stimulation frequency was 2 Hz, the intensity was 2 mA, continuous wave, 20 min daily, and continuous intervention for 14 d. After the intervention, the expression of CREB and p-CREB protein in hippocampus, cortex and cerebellum were detected by immunohistochemistry. Result ①The CREB expression in hippocampus of the blank group was significantly higher than that in cortex (P<0.05) and cerebellum (P<0.001).The CREB expression in cortex of the non-acupoint group and the GV1-acupoint group was significantly higher than that in hippocampus (P<0.05, P<0.01) and cerebellum (P<0.05).②The expression of p-CREB and p-CREB rate of the blank group in cerebellum were significantly higher than those in cortex (P <0.01, P <0.001) and hippocampus (P <0.05, P <0.001).The expression of p-CREB in cerebellum of non-acupoint group was significantly higher than that in hippocampus (P <0.01).Compared with cortex (P <0.01) and hippocampus (p <0.05), p-CREB rate in cerebellum of non-acupoint group was significantly increased.There was no significant difference between the three brain regions in the GV1-acupoint group (P > 0.05).Contour analysis results are as follows:①The results of parallel contour analysis of CREB showed P < 0.05, indicating that the overall contour of the three brain regions were not parallel, that is, the linkage changes of the brain regions were not consistent.②Horizontal contour analysis of p-CREB and p-CREB rate showed P < 0.01. The results suggested that the linkage changes of the three brain regions were consistent, and the degree of change was consistent among the groups, but the protein expression in each group was not equal, among which cerebellum was the highest. Conclusion Based on the contour analysis, this study can initially observe the multi-brain linkage effect of acupuncture, and the acupuncture stimulation may affect the multi-brain effect of FMR1 knockout mice by affecting CREB expression.