益肾养髓方对脊髓型颈椎病大鼠线粒体自噬及血脊髓屏障功能的影响  

Effects of Yishen Yangsui Formula on mitochondrial autophagy and blood-spinal cord barrier function in rats with cervical spondylotic myelopathy

作　　者：许博 杨博文 秦晓宽 张典 严正 赵泽龙 张博宇 金哲峰[1] 魏戌[1] 朱立国[1,3] 陈忻 银河[1] XU Bo;YANG Bowen;QIN Xiaokuan;ZHANG Dian;YAN Zheng;ZHAO Zelong;ZHANG Boyu;JIN Zhefeng;WEI Xu;ZHU Liguo;CHEN Xin;YIN He(Wangjing Hospital of China Academy of Chinese Medical Sciences,Beijing 100102,China;Graduate School of Beijing University of Chinese Medicine,Beijing 100029,China;Beijing Key Laboratory of Traditional Chinese Medicine Orthopaedic Technology,Beijing 100700,China)

机构地区：[1]中国中医科学院望京医院,北京100102 [2]北京中医药大学研究生院,北京100029 [3]北京中医正骨技术重点实验室,北京100700

出　　处：《中华中医药杂志》2024年第11期5838-5843,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金青年科学基金项目(No.81904232,No.82205151);国家中医药管理局中医药创新团队及人才支持计划项目(No.ZYYCXTD-C-202003);中国中医科学院望京医院基础研究苗圃培育计划课题(No.WJYY-YJKT-2022-02)。

摘　　要：目的:观察益肾养髓方对脊髓型颈椎病大鼠血脊髓屏障(BSCB)及其中线粒体自噬的影响。方法:27只SD大鼠随机分为假手术组、模型组、益肾养髓方组,每组9只。采用吸水膨胀材料构建大鼠脊髓慢性受压模型。利用BBB评分、伊文思蓝(EB)染色、荧光显微镜观察、免疫荧光多重标记法和Western blot等技术,评估大鼠的运动功能、BSCB通透性、内皮细胞损伤和线粒体自噬的变化。结果:与模型组比较,益肾养髓方组大鼠在第3、7、14天BBB评分显著升高(P<0.05)。模型组大量神经元细胞受损,而益肾养髓方组神经元形态和数量有一定恢复趋势。益肾养髓方组Claudin-5、ZO-1、Occludin蛋白较模型组表达水平显著升高(P<0.01),EB渗出量显著下降(P<0.05)。益肾养髓方组LC3标记的自噬小体与Hsp60标记的线粒体有共定位信号(P<0.05),且LC3和Hsp60的荧光标记数量较模型组更多(P<0.05)。结论:益肾养髓方能显著调控线粒体自噬功能,减轻线粒体受损的程度,并有效缓解大鼠BSCB的损伤。Objective:To investigate the effects of Yishen Yangsui Formula(YYF)on the blood-spinal cord barrier(BSCB)and mitochondrial autophagy in a rat model of chronic spinal cord compression.Methods:Twenty-seven SD rats were randomly assigned to the sham group,model group,and YYF group,with 9 rats in each group.The chronic spinal cord compression model was established using water swelling material.Motor function,BSCB permeability,endothelial cell injury,and mitochondrial autophagy were assessed by the BBB score.This was assessed using Evans Blue staining,fluorescence microscopy,immunofluorescence multiple labeling,and Western blot.Results:The BBB score in the YYF group significantly highered at the 3rd,7th,14th day than that in the model group(P<0.05).Many neurons in the model group were damaged.However,the YYF group showed a certain trend in neuronal recovery in terms of morphology and quantity.The expression levels of Claudin-5,ZO-1,and Occludin proteins in the YYF group were significantly higher than these in the model group(P<0.01),and the exudation of Evans blue was significantly decreased(P<0.05).The co-localization signals between autophagosomes labeled by LC3 and mitochondria labeled by Hsp60 in the YYF group(P<0.05),which was associated with a higher number of fluorescence labels of LC3 and Hsp60 than the model group(P<0.05).Conclusion:YYF can significantly regulate the function of mitophagy,reduce the degree of mitochondrial damage,and effectively alleviate the damage of BSCB in rats.

关 键 词：脊髓型颈椎病 血脊髓屏障 内皮细胞 线粒体 益肾养髓方 

分 类 号：R285.5[医药卫生—中药学]