方氏头针对血管性痴呆大鼠海马CA 1区星形胶质细胞凋亡的影响

目的:观察方氏头针对血管性痴呆大鼠海马CA 1区星形胶质细胞凋亡的影响,探讨方氏头针治疗血管性痴呆的机制。方法:健康雄性SD大鼠随机分为正常组、假手术组、模型组、方氏头针组,每组10只。采用改良Pulsinelli四血管阻断法制备血管性痴呆大鼠模型。方氏头针组针刺头部"倒脏上焦"(双侧)、"记忆"(双侧)、"思维"穴,每日1次。10d后,应用Morris智能水迷宫测试各组大鼠学习记忆能力的变化,采用免疫荧光双重标记技术检测各组大鼠海马CA 1区星形胶质细胞特异性标记物胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)与抑凋亡因子B细胞淋巴瘤/白血病-2(B cell lymphoma/leukemia-2,Bcl-2)共表达的阳性细胞数目,采用Western blot技术检测海马星形胶质细胞抑凋亡因子Bcl-2蛋白的表达。结果:水迷宫实验结果显示,平均逃避潜伏期及2min内首次跨越原象限平台的时间比较,模型组明显长于正常组及假手术组(P0.01),方氏头针组明显短于模型组(P0.01)。2min内跨越原象限平台次数比较,模型组明显少于正常组及假手术组(P0.01),方氏头针组明显多于模型组(P0.01)。与正常组及假手术组比较,模型组海马CA 1区GFAP/Bcl-2共表达的阳性细胞数明显降低,Bcl-2蛋白表达明显下降(P0.01);与模型组比较,方氏头针组GFAP/Bcl-2共表达阳性细胞数显著增高,Bcl-2蛋白表达升高(P0.01)。结论:方氏头针可显著上调血管性痴呆大鼠海马CA 1区星形胶质细胞抑凋亡因子Bcl-2的表达,从而有效改善血管性痴呆大鼠的学习记忆能力。

Effects of Scalp-acupuncture on Astrocyte Apoptosis in Hippocampal CA 1 Region in Rats with Vascular Dementia

Objective To observe the effect of scalp-acupuncture intervention on the expression of Bcl-2of astrocytes in the hippocampus in vascular dementia(VD)rats,so as to reveal its underlying mechanism in improving VD.Methods A total of40 male SD rats were randomly divided into normal control,sham-operation,VD model,and scalp-acupuncture groups(n=10in each group).The VD model was established by using modified Pulsinelli 4-vessel-occlusion method.Scalp-acupuncture was applied to bilateral"Daozang Shangjiao"and bilateral"Jiyi",and"Siwei"acupoints according to FANG's scalp-acupuncture therapy for 30 min,once daily for 10 days.The rat's learning-memory ability was detected by Morris water maze tests,and the expression of apoptosis related proteins Bcl-2of astrocytes in the hippocampus was detected by immunofluorescence double labeling technique and Western blot,respectively.Results Morris water maze tests showed that the average escape latency of place navigation test and the duration of the first platform-site crossover(space probing test)were significantly increased in the model group compared with the normal control group(P<0.01),and the number of platform-site crossovers in 2min was markedly decreased in the model group(P<0.01).The increased average escape latency and the increased duration of first platform-site crossover were markedly shortened in the scalp-acupuncture group in comparison with the model group(P<0.01),and the number of the platform crossovers in 2min was significantly increased in the scalp-acupuncture group(P<0.01).The expression of Bcl-2protein and the number of GFAP/Bcl-2-labeled cells of hippocampal CA 1region in the model group were remarkably decreased compared with the normal control group(P<0.01).After scalp-acupuncture intervention,the decreased Bcl-2protein expression level and the number of double-labeled GFAP/Bcl-2cells were significantly up-regulated in the scalp-acupuncture group compared with the model group(P<0.01).No significant differences were found between the normal control and sham-operation groups in the escape latency,duration of the first platform-site crossover,number of platform crossovers in 2 min,number of GFAP/Bcl-2double labeled cells and the expression level of hippocampal Bcl-2protein(P>0.05).Conclusion Scalp-acupuncture can effectively regulate the expression of apoptosis related proteins Bcl-2of astrocytes in the CA 1region of hippocampus in rats with VD,which may contribute to its effect in the improvement of VD.