针刺联合栀子苷对糖尿病复合脑缺血大鼠脑内小胶质细胞激活及核转录因子-κB表达的影响

Influence of Electroacupuncture plus Jasminoidia Intervention on Expression of OX-42 and Nuclear Factor-kappa B in Hippocampus and Amygdale in Cerebral Ischemia plus Diabetes Rats

目的:探讨针刺联合栀子苷的脑保护机制。方法:雄性Wistar大鼠,随机分为正常组、模型组、药物组、针刺组、针药结合组,每组12只。链脲佐菌素腹腔注射制作糖尿病模型,双侧颈总动脉阻断法制造脑缺血再灌注模型。药物组大鼠予以栀子苷(25mg/kg)生理盐水溶液灌胃,每日1次,连续治疗1周;电针组大鼠选取"百会"和"大椎"两穴,每次电针20min,隔日1次,共10次;针药结合组大鼠接受栀子苷治疗的同时,也接受电针治疗。Morris水迷宫行为实验测试大鼠学习记忆状况;免疫组化法检测海马及杏仁核内小胶质细胞标志物OX-42和核转录因子-κB(NF-κB)免疫活性的变化。结果:电针、药物以及针药结合3个治疗组治疗后,动物在撤台后于靶象限游泳路程的百分比和游泳时间的百分比较模型组明显增加(P<0.05)。模型组大鼠海马与杏仁核OX-42阳性标记小胶质细胞数量与吸光度明显高于正常组(P<0.01),治疗后明显降低(P<0.05),3个治疗组间差异无统计学意义(P>0.05)。模型组大鼠海马和杏仁核NF-κB阳性标记细胞数量与吸光度显著高于正常组(P<0.05),治疗后明显降低(P<0.05),3个治疗组间差异无统计学意义(P>0.05)。结论:针刺和栀子苷均通过NF-κB-小胶质细胞信号通路,抑制糖尿病复合脑缺血模型大鼠脑内小胶质细胞的活性,从而缓解糖尿病复合脑缺血模型大鼠的学习记忆障碍。针刺加栀子苷复合治疗与单纯针刺或药物治疗效果相同。

Objective To observe the changes of expression of OX-42 and nuclear factor-kappa B （NF-κB） transporter factor in the hippocampus and amygdale in diabetes and cerebral ischemia （CI） rats with learning and memory impairment after combined treatment with electroacupuncture （EA） plus Jasminaidia, so as to explore their mechanism underlying cerebral protection. Methods Sixty male Wistar rats were randomly divided into control, model, medication, EA, and EA＋medication groups （12 rats/group）. The diabetes mellitus model was established by intraperitoneal injection of Streptozotocin （60 mg/kg, for 3 days.and the CI model made by repeated occlusion and reperfusion of the bilateral carotid arteries. The rats＇ lear-ning-memory ability was measured via Morris water maze tasks. Rats of the medication group were fed with Jasminaidia （25 mg/kg）, once a day for one week, and those of the EA group administrated with EA stimulation of ＂Baihui＇（GV 20） and ＂Dazhui＂（GV 14）, once every other day for 20 days. For rats of the EA＋medication group, both EA and Jasminaidia were given the same to those mentioned above. Three weeks later, changes in the number and optical density of OX-42 （microglial cell mar-kers）-positive and NF-κB-positive cells in the hippocampus and amygdale were examined by immunohistochemistry. Results After the treatment, the diabetes＋CI induced decrease of percentages of swimming distance and swimming time in the target quadrant were obviously increased in the EA, medication and EA＋medication groups （P〈0. 05）, suggesting an im- provement of the rats＇ memory ability. The number of OX-42 positive microglial cells and NF-κB-positive cells and their immu- noactivity in the hippocampus and amygdale were significantly higher in the diabetes plus CI model rats than in the control rats（P〈0.01 ,P%0.05）. After the treatment, the number and expression levels of OX 42 marked microgliacytes and NF-κB-positive cells were significantly decreased in the EA, medication and EA medication groups compared with the model group （P〈0.05）. No significant differences were found among the 3 treatment groups in the number of OX-42- and NF-κB-positive cells and their expression levels （P〉0.05）. Conclnsion EA combined with Jasminoidia intervention can improve the learn- ing-memory ability and inhibit the microglial cell activity in diabetes＋CI rats probably via down-regulation of NF κB signaling pathway.