摘要
目的探究肌萎缩性侧索硬化症(amyotrophic lateral sclerosis,ALS)患者血清外泌体和外周血CD14^(++)单核细胞的相互作用及ALS可能的发病机制。方法选择2013年1月至2019年12月于四川遂宁市中心医院就诊的ALS患者90例作为ALS组,同期随机选择健康志愿者30例作为对照组。用ALS患者和健康人血清外泌体刺激健康人和ALS患者CD14^(++)单核细胞,分析单核细胞因子的分泌功能。从HEK293细胞的条件培养基中制备Luc、TDP(WT)-Luc以及ALS突变型TDP(M337V)-Luc的外泌体,分别刺激ALS患者/健康人CD14^(++)单核细胞,分析单核细胞的吞噬功能和细胞因子的分泌功能。结果健康人CD14^(++)单核细胞的分泌功能与外泌体供体相关性不大,不同组别外泌体培养下的健康人CD14^(++)单核细胞分泌的促炎细胞因子水平无明显差异(P>0.05);而不同组别外泌体培养下的ALS组CD14^(++)单核细胞分泌的促炎细胞因子表达水平存在显著差异,ALS患者单核细胞趋化蛋白-1(monocyte chemotactic protein 1,MCP-1)、白细胞介素(interleukin,IL)-6、IL-13的表达水平较对照组明显升高[(38.41±2.33)pg/mL比(35.15±2.42)pg/mL,(0.35±0.03)pg/mL比(0.29±0.02)pg/mL,(4.43±0.35)pg/mL比(4.15±0.23)pg/mL,t值分别为2.143,1.537和1.574,P值均<0.05],差异具有统计学意义。对人CD14^(++)单核细胞对外泌体的摄取量与外泌体中的TDP-43的关系进行分析发现,健康人CD14^(++)单核细胞对含有TDP-43的外泌体摄取效率为Luc外泌体的10倍,但是对ALS患者突变型TDP(M337V)-Luc的外泌体摄取量则降低,提示ALS患者CD14^(++)单核细胞吞噬功能受损。此外,相对于Luc外泌体,TDP(WT)-Luc外泌体提高了健康人和ALS患者CD14^(++)单核细胞IL-6,IL-10,IL-1b和MCP-1的分泌功能,TDP(M337V)-Luc外泌体则抑制其分泌功能。结论 ALS中突变的TDP-43可能通过负载的外泌体诱导单核细胞,使单核细胞的激活功能失调,吞噬功能受损,导致更多的TDP-43聚集在中枢�
Objective To investigate the effects of serum exosomes on peripheral blood CD14^(++)monocytes in patients with amyotrophic lateral sclerosis(ALS)and the possible pathogenesis of amyotrophic lateral sclerosis.Methods Total of ninety patients with amyotrophic lateral sclerosis were selected in Sichuan Suining Central Hospital and West China Hospital from January 2013 to December 2019(ALS group).At the same time,thirty healthy volunteers were randomly selected(control group).Serum exosomes from ALS patients and healthy people,were used to stimulate the CD14^(++)monocytes to analyze the secretion function of monocyte cytokines.The exosomes of Luc,TDP(WT)-Luc and ALS mutant TDP(M337V)-Luc were prepared from the conditioned medium of HEK293 cells,to stimulate CD14^(++)monocytes in healthy people or ALS patients.The phagocytic function of monocytes and the secretory function of cytokines were analyzed.Results There was no significant correlation between the secretion function of CD14^(++)monocytes and the donors,and no significant differences were found in the levels of inflammatory cytokines secreted by CD14^(++)monocytes in healthy people(P>0.05).The levels of proinflammatory cytokines secreted by CD14^(++)monocytes were significantly different in ALS group cultured with different exosomes.The expression level of monocyte chemoattractant protein-1(MCP-1),interleukin(IL)-6 and IL-13 in ALS patients were significantly higher than those in controls[(38.41±2.33)pg/mL vs(35.15±2.42)pg/mL,(0.35±0.03)pg/mL vs(0.29±0.02)pg/mL,(4.43±0.35)pg/mL vs(4.15±0.23)pg/mL,t values were 2.143,1.537 and 1.574 respectively,all P values<0.05].The relationship between the uptake of exosomes by CD14^(++)monocytes and TDP-43 in exosomes were analyzed.The result showed that the uptake efficiency of CD14^(++)monocytes to exosomes containing TDP-43 was 10 times higher than that of Luc exosomes,but the exosomes uptake of mutant TDP(m337v)-Luc in ALS patients was decreased,indicating that the phagocytic function of CD14^(++)monocytes in ALS p
作者
张运伟
喻明
李琳林
王兰
Zhang Yunwei;Yu Ming;Li Linlin;Wang Lan(Neurology Department,Suining Central Hospital,Suining 629000,China;Neurology Department,West China Hospital,Chengdu 610041,China)
出处
《国际免疫学杂志》
CAS
2021年第1期23-28,共6页
International Journal of Immunology
