益肾养髓方对脊髓型颈椎病大鼠脊髓中细胞凋亡的影响  

Effect of Yishen Yangsui Formula on Apoptosis in the Spinal Cord of Rats with Cervical Spondylotic Myelopathy

作　　者：张典 朱立国[2,3,4] 尹逊路 杨博文 秦晓宽 金哲峰 许博[2] 张博宇 陈忻 银河[2] ZHANG Dian;ZHU Liguo;YIN Xunlu;YANG Bowen;QIN Xiaokuan;JIN Zhefeng;XU Bo;ZHANG Boyu;CHEN Xin;YIN He(Beijing University of Chinese Medicine,Beijing 100029,China;Department of Spine,Wangjing Hospital of China Academy of Chinese Medical Sciences,Beijing 100102,China;Beijing Institute of Orthopedics of Integrated Traditional Chinese and Western Medicine,Beijing 100700,China;Beijing Key Laboratory of Bone Setting Technology of Traditional Chinese Medicine,Beijing 100700,China)

机构地区：[1]北京中医药大学,北京100029 [2]中国中医科学院望京医院脊柱二科 [3]北京市中西医结合骨伤研究所 [4]中医正骨技术北京市重点实验室

出　　处：《中国中医骨伤科杂志》2024年第1期1-6,共6页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：国家自然科学基金项目(82205151);中国中医科学院望京医院基础研究苗圃培育计划课题(WJYY-YJKT-2022-02);中国中医科学院望京医院自主选题专项课题(WJYY-ZZXT-2022-08);中国中医科学院基本科研业务费优秀青年科技人才培养专项(ZZ16-YQ-025)。

摘　　要：目的:观察益肾养髓方对脊髓型颈椎病大鼠脊髓中细胞凋亡及Fas/FasL-Caspase-8/-3通路的影响。方法:将大鼠随机分配至假手术组、模型组及益肾养髓方高、中、低浓度组,采用吸水膨胀材料压迫脊髓的方法建立脊髓型颈椎病动物模型,术后2周分别给予生理盐水或相应中药水煎剂灌胃。采用Basso Beatie Bresmahan(BBB)评分法测量大鼠运动功能,TUNEL染色评估细胞凋亡,RT-qPCR和Western Blot检测Fas、FasL、Caspase-8及Caspase-3表达情况。结果:造模术后2周,益肾养髓方各组及模型组BBB评分显著低于假手术组(P<0.05)。术后6周,益肾养髓方中浓度组BBB评分均显著优于模型组(P<0.05)。TUNEL染色显示,益肾养髓方各组细胞凋亡率均显著低于模型组(P<0.05)。术后6周,RT-qPCR检测显示,益肾养髓方高、中、低浓度组Caspase-3 mRNA表达水平均低于模型组(P<0.05),中、低浓度组Caspase-8 mRNA表达水平均低于模型组(P<0.05),高、中浓度组Fas mRNA表达水平均低于模型组(P<0.05),低浓度组FasL mRNA表达水平低于模型组(P<0.05)。Western Blot检测显示,益肾养髓方高、中、低浓度组Caspase-3蛋白相对表达量低于模型组(P<0.05),益肾养髓方中浓度组Caspase-8蛋白相对表达量低于模型组(P<0.05),中、低浓度组Fas及FasL蛋白相对表达量低于模型组(P<0.05)。结论:益肾养髓方可通过抑制Fas/FasL-Caspase-8/-3信号通路减少受损脊髓区域的细胞凋亡,进而改善脊髓型颈椎病。Objective:To investigate the effects of Yishen Yangsui formula on apoptosis and Fas/FasL-Caspase-8/-3 pathway in spinal cord of rats with cervical spondylotic myelopathy(CSM).Methods:Rats were randomly assigned to sham operation group,model group,high,middle and low dosage groups of Yishen Yangsui formula.The CSM rat model was established by the chronic compression of spinal cord with water swelling material.2 weeks after the operation,normal saline or corresponding chinese herbal decoction were given for gavage.Basso Beatie Bresmahan(BBB)score was used to evaluate the spinal cord function of rats.TUNEL staining was used to evaluate apoptosis.RT-qPCR and Western Blot were used to detect the expression of Fas,FasL,Caspase-8 and Caspase-3.Results:2 weeks after modeling,the BBB score of each group given Yishen Yangsui formula and model group were significantly lower than those of sham operation group(P<0.05).6 weeks after surgery,the BBB score of middle-dosage group of Yishen Yangsui formula was higher than that of model group,with statistical differences(P<0.05).TUNEL staining showed that the apoptosis rate of groups of Yishen Yangsui formula was lower than that of model group,with statistical differences(P<0.05).6 weeks after modeling,RT-qPCR showed that the expression levels of Caspase-3 mRNA in the high,middle and low dosage groups of the Yishen Yangsui formula were lower than those in the model group(P<0.05),the expression levels of Caspase-8 mRNA in the middle-and low-dosage groups were lower than those in the model group(P<0.05),the expression levels of Fas mRNA in the high and middle dosage groups were lower than those in the model group(P<0.05),and the expression levels of FasL mRNA in the low dosage group were lower than those in the model group(P<0.05).The Western Blot showed that the relative expression of Caspase-3 in the high,middle and low dosage groups of the Yishen Yangsui formula was lower than that of the model groups(P<0.05),the relative expression of Caspase 8 in the middle dosage group of Yishe

关 键 词：益肾养髓方 脊髓型颈椎病 退行性颈脊髓病 神经炎症 细胞凋亡 

分 类 号：R-33[医药卫生]