摘要
目的探讨动脉粥样硬化疾病进展过程中辅助性T细胞17（Thl7）和调节性T细胞（Treg）之间功能失衡的机制。方法利用载脂蛋白E缺陷型（apoE-/-）小鼠建立动脉粥样硬化疾病模型，随机数字表法分成4组，每组10只，正常饮食饲养至相应周龄，即6、12、24和48周龄，同周龄C57／B6小鼠作为对照组；用流式细胞术检测Th17和Treg细胞及树突状细胞（DC）的数量和表型；用酶联免疫吸附法检测细胞因子白细胞介素（IL）-6、IL-17A和转化生长因子（TGF）-B1血清水平；用混合淋巴细胞反应检测Treg细胞抑制功能。结果随着周龄的增加，apoE-/-小鼠脾脏Thl7和Treg细胞占总CIM’T细胞的比例均显著增加，6周龄apoE-/-小鼠Thl7和Treg占总CI4＋T细胞的比例分别为1．00％和8．08％，48周龄apoE“小鼠Thl7和Treg占总CIM’T细胞的比例分别为3．14％和27．80％，而不同周龄C57／B6对照小鼠脾脏中的Thl7／Treg比值始终保持稳定。虽然Thl7和Treg细胞占总CD4＋T细胞的比例明显增加，但Thl7／Treg比值保持稳定，各组问差异无统计学意义（P〉0．05）；血清IL-17A水平随周龄增加而增加[6周龄：（87±15）pgJml，48周龄：（191±26）pg/ml]，但TGF-β1水平未发生显著改变；脾脏DC的数量也保持稳定，但显著高水平表达CD40、CD80、CD86和MHC-11类分子，IL-6的水平也显著上升[从6周龄的（43±5）pg/ml上升到48周龄的（104±11）pg/m1]；体外混合淋巴细胞反应结果显示IL-6下调Treg细胞的抑制能力。结论动脉粥样硬化疾病进展中，DC被过度活化并分泌前炎症因子IL-6，抑制Treg细胞功能的发挥，导致Thl7和Treg细胞功能欠衡可能是疾病进展的机制之一。
Objective To investigate the role of the helper T cells （Th） 17/Treg cell imbalance on the development of atherogenesis in apo E knockout mice. Methods Apo E -/- mice were examined at age of 6,12,24 and 48 weeks （n = 10 each）. Age matched C57/B6 mice served as controls. The number of Thl7, Treg and dendritic cell （DC） was detected by flowcytometry. The levels of interleukin（IL）-6, IL- 17A and transforming growth factor（ TGF） -[31 were detected by ELISA. The suppression ability of Treg was evaluated by mixed lymphocyte reaction. Results With increasing ages, the frequencies of Thl7 and Treg in CD4 ~ T cells were increased（Thl7 ratio from 1.00% to 3.14% ; Treg ratio from 8.08% to 27.80% ） and the level of IL-17A was up-regulated [ from （87 ＋ 15） pg/ml to （191 ＋ 26） pg/ml], but the rate of Thl7/ Treg cell and the level of TGF-[31 remained stable during atherogenesis in apo E knockout mice. Furthermore, the phcnotype of splenic DC was matured and the blood level of IL-6 was up-regulated [ from （43 ＋ 5 ） pg/ml to （ 104 ＋ 11 ） pg/ml] with aging in apo E / mice. Addition of IL^6 to T cells reversed the ability of Treg to suppress the proliferation of effective T cells. Conclusion DC overactivation, subsequent increased secretion of IL-6, inhibition of Treg cell function and the Thl7/Treg cell imbalance play key roles on the atherogenesis in apo E-/- mice.
出处
《中华心血管病杂志》
CAS
CSCD
北大核心
2013年第5期416-421,共6页
Chinese Journal of Cardiology
基金
浙江省自然科学基金（Y2100526）
浙江省科技厅科研项目（2009C33149）
浙江省医药卫生科技计划项日（20098123）
杭州市科技发展计划项目（20110733Q14）
