电针对缺血性脑卒中模型大鼠热休克蛋白HSP60表达的影响

刘梅芳 杨光至 张霞辉 冯斯峰 徐连杰 周以皓 施静

摘要：目的 观察针灸对脑梗死大鼠HSP60、Caspase-3表达规律，初步探讨针灸对缺血性脑卒中大鼠神经细胞凋亡保护的作用机制。方法 采用大脑中动脉线栓诱导脑缺血再灌注损伤模型，随机假手术组、模型组、电针穴位组、电针非经非穴和艾灸穴位组。电针或艾灸干预后，术后0、12h评估大鼠的神经功能缺陷评分和姿势反射评分;荧光定量PCR检测HSP60和Caspase-3 mRNA的表达水平。结果 电针穴位组、艾灸穴位组大鼠神经功能均优于模型组，差异有统计学意义（P<0.05）;与模型组比较，电针穴位组大鼠HSP60 mRNA显著增加，差异有显著性意义（P<0.05）;而Caspase-3 mRNA表达无明显变化。结论 电针对大鼠脑缺血再灌注损伤神经功能有改善作用，其机制可能与上调脑组织中 HSP60表达，介导抗神经细胞凋亡有关。

关键词：缺血性脑卒中;HSP60;Caspase-3;神经细胞凋亡

中图分类号：R246 文献标志码：A 文章编号：1007-2349（2021）05-0069-04

【Abstract】Objective： To study the determination method of polysaccharides in Aconitum racemulosum and to establish a UV spectrophotometric method for the determination of polysaccharides from Aconitum Racemulosum， and to investigate the influence of different producing areas and different processing methods on the content of polysaccharides. Methods： The impurities such as reducing sugars were removed by 80% ethanol reflux and then the polysaccharides were extracted with hot water. The phenol-sulfuric acid colorimetry was used to measure absorbance value at 488 nm wavelength by ultraviolet-spectrophotometry to obtain the content of polysaccharides. . Results： The polysaccharide of Aconitum racemulosum was calculated as anhydrous glucose and showed a good linear relationship in the range of 0.003 to 0.015 mg/mL（R=0.9999）， and the recovery rate was 99.35%-99.85%（n=6）， and the RSD value was 0.18 %. Conclusion： This method is accurate， reliable， and reproducible， which is suitable for the determination of polysaccharide content in Aconitum racemulosum. There are certain differences in the polysaccharide content in different batches of medicinal materials from different origins. The processing method has no significant effect on the polysaccharide content of Aconitum racemulosum.

【Key words】Aconitum racemulosum; Polysaccharide; Ultraviolet Spectrophotometry; Producing Area; Processing

中風有着极高的发病率、患病率、死亡率、致残率和复发率。世界范围内，发病率平均为140～200/10万[1]。缺血性脑中风仍是当今疾病中主要的致残、致死原因，严重威胁人类的健康，并给家庭和社会带来了很大的负担，因此，探讨其发病机制，寻找有效的治疗方法一直是医学工作者研究的热点。缺血性脑损伤疾病发生与神经细胞凋亡密切相关，凋亡发生的重要原因之一是线粒体凋亡传导通路的激活。线粒体通路激活后，继而启动Caspase-9激活，继续激活Caspase-3，从而导致Caspase的级联反应，最终促使细胞凋亡的发生[2-3]。HSP60作为热休克蛋白家族中重要成员之一，在细胞凋亡过程中有其较强关联性[4]。

电针疗法是将针刺和电刺激相结合，通过脉冲电流作用于神经、肌肉引起神经兴奋性传导，消除神经纤维间水肿和神经滋养血管的痉挛，对脑卒中具有较好的临床疗效。……