温肾通督方对脊髓损伤后微血管内皮细胞吞噬髓鞘碎片的影响  

Effects of Wenshen Tongdu Decoction on phagocytosis of myelin debris by microvascular endothelial cells after spinal cord injury

作　　者：吴承杰 郭杨[1] 石磊[2] 陈思娴 潘娅岚 刘孟敏 王礼宁 马勇[1,2] WU Cheng-jie;GUO Yang;SHI Lei;CHEN Si-xian;PAN Ya-lan;LIU Meng-min;WANG Li-ning;MA Yong(Laboratory of New Techniques of Restoration&Reconstruction,Institute of Traumatology&Orthopedics,Nanjing University of Chinese Medicine,Nanjing 210023,China;School of Chinese Medicine,School of Integrated Chinese and Western Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,China;Laboratory of Chinese Medicine Nursing Intervention for Chronic Diseases,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学骨伤修复与重建新技术实验室,南京210023 [2]南京中医药大学中医学院·中西医结合学院,南京210023 [3]南京中医药大学慢性病中医护理干预实验室,南京210023

出　　处：《中华中医药杂志》2023年第5期2073-2078,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(N o.81973885);南京中医药大学自然科学基金青年项目(No.NZY81704100);江苏省高校自然科学基金项目(No.21KJB360006);江苏高校中西医临床医学品牌专业建设工程资助项目(二期)(No.2020PPZXL261);2022江苏省研究生科研与实践创新计划(No.KYCX22_1942)。

摘　　要：目的:探究温肾通督方对脊髓损伤后微血管内皮细胞(MECs)吞噬髓鞘碎片的影响及其作用机制。方法:采用改良A11en’s法制备脊髓损伤小鼠模型,1、3、7d后通过HE染色和尼氏染色观察损伤脊髓的病理改变。利用ELISA法检测损伤脊髓和血清中MCP-1、IL-6、IL-1β、TNF-α的表达水平,通过3D激光共聚焦观察损伤脊髓中MECs吞噬髓鞘碎片情况,采用多重免疫荧光染色检测脊髓损伤后MECs中HDAC6、LC3B、NLRP3表达。结果:与模型组比较,温肾通督方明显减少脊髓损伤后炎症细胞的浸润和神经元的固缩坏死;显著降低脊髓和血清中MCP-1、IL-6、IL-1β、TNF-α的水平(P<0.01,P<0.05);促进MECs吞噬髓鞘碎片,并能下调HDAC6、NLRP3表达,上调LC3B表达(P<0.01)。结论:温肾通督方可能通过抑制HDAC6促进MECs吞噬髓鞘碎片,并能同时阻断NLRP3通路的信号转导,共同改善炎性微环境,促进脊髓损伤修复。Objective:To explore the effects of Wenshen Tongdu Decoction on phagocytosis of myelin debris by microvascular endothelial cells(MECs)after spinal cord injury and its mechanism.Methods:The mouse model of spinal cord injury was established by modified Allen’s method.The pathological changes of injured spinal cord were observed by HE staining and Nissl staining after 1,3 and 7 days.The expression levels of MCP-1,IL-6,IL-1βand TNF-αin injured spinal cord and serum were detected by ELISA method.The phagocytosis of myelin debris by MECs in injured spinal cord was observed by 3D laser confocal scanning.The expression of HDAC6,LC3B and NLRP3 in MECs after spinal cord injury was detected by multiple immunofluorescence staining.Results:Compared with the model group,Wenshen Tongdu Decoction significantly reduced the infiltration of inflammatory cells and pyknosis necrosis of neurons after spinal cord injury;Wenshen Tongdu Decoction could significantly reduce the levels of MCP-1,IL-6,IL-1βand TNF-αin spinal cord and serum(P<0.01,P<0.05);Wenshen Tongdu Decoction could promote MECs to phagocytize myelin debris,downregulate the expression of HDAC6 and NLRP3,and up-regulate the expression of LC3B(P<0.01).Conclusion:Wenshen Tongdu Decoction may promote the phagocytosis of myelin debris by MECs by inhibiting HDAC6,and block the signal transduction of NLRP3 pathway at the same time,jointly improve the inflammatory microenvironment and promote the repair of spinal cord injury.

关 键 词：温肾通督方 脊髓损伤 微血管内皮细胞 髓鞘碎片 炎症反应 

分 类 号：R285.5[医药卫生—中药学]