海南医科大学学报

目的:探讨针刺对阿尔茨海默病(AD)大鼠海马神经元内质网应激及凋亡的影响。方法:36只Wistar大鼠随机分为空白组、模型组、假手术组和针刺组,模型组和针刺组通过双侧海马CA1区注射Aβ_1-42建立AD模型,假手术组以等体积的无菌双氧水注射,空白组不做处置。针刺组于造模后2 d起给予针刺干预,空白组、模型组和假手术组不予针刺仅背侧位捆绑固定,1次/d,连续治疗14 d。疗程结束后通过HE染色观察大鼠海马CA1区神经元形态,Tunel法检测海马CA1区神经元凋亡水平,免疫组化法检测海马CA1区Aβ_1-42、GRP78蛋白的表达,Western blot法检测海马CA1区GRP78、Caspase-12蛋白的水平。结果:与空白组大鼠相比,模型组大鼠海马CA1区神经元细胞明显减少,凋亡水平、Aβ_1-42、GRP78和Caspase-12蛋白表达水平明显增高,差异具有统计学义(P<0.01)。与模型组相比,针刺组大鼠海马CA1区神经元形态较模型组明显改善,细胞数量明显增多,凋亡水平、Aβ_1-42、GRP78和Caspase-12蛋白的表达明显降低,差异具有统计学义(P<0.01)。结论:针刺治疗能够通过抑制内质网应激和细胞凋水平,进而改善海马CA1区神经元形态和抑制Aβ_1-42的堆积。

Objective:To investigate the effects of acupuncture on endoplasmic reticulum stress and apoptosis in hippocampal neurons of rats with Alzheimer's disease(AD). Methods: 36 Wistar rats were randomly divided into the control group, model group, sham-operated group, and acupuncture group. The AD model was replicated by injecting β-amyloid protein 1-42 solution into bilateral hippocampal CA1 area in model group and acupuncture group. The sham-operated group was injected with the same amount of sterile hydrogen peroxide while the control group was not treated. Rats in the acupuncture group were treated with cluster needling of scalp point therapy from 2 d after model establishment once a day for 14 consecutive days, while rats in the control group, model group and sham-operated group were bundled and fixed in the dorsal position instead. After treatment, the morphology of hippocampal CA1 neurons was observed by HE staining, the apoptotic level of hippocampal CA1 neurons was detected by Tunel method, the expression of Aβ_1-42 and GRP78 proteins in hippocampal CA1 region was detected by immunohistochemistry, and the levels of GRP78 and Caspase-12 proteins in hippocampal CA1 region were detected by Western blot. Results:Compared with the control group, the neurons in hippocampal CA1 area in the model group decreased significantly, and the apoptotic rate, the expression of Aβ_1-42, GRP78 and Caspase-12 protein increased significantly(P<0.01). Compared with the model group, the morphology of hippocampal CA1 neurons in the acupuncture group was significantly improved, the number of neurons was significantly increased, the apoptotic rate, the expression of Aβ_1-42, GRP78 and Caspase-12 protein were significantly decreased, the difference was statistically significant(P<0.01). Conclusions:Scalp acupoint cluster needling therapyimproves the morphology of hippocampal CA1 neurons and inhibit the accumulation of Aβ_1-42 by inhibiting endoplasmic reticulum stress and cell apoptosis.