CAJ | 학술논문

目的研究双链RNA(double-stranded RNA,dsRNA)和咪喹莫特联合刺激是否具有协同效应.方法在Toll样受体3(TLR3)的特异性刺激剂dsRNA、TLR7的特异性刺激剂咪唪莫特(R837)单独刺激或二者联合刺激条件下,检测BALB/c×C57BL/6和NH细胞缺陷的非肥胖型糖尿病(NOD)×C57BL/6小鼠胚胎吸收率.采用小鼠体内注射上述刺激剂的方法 ,流式细胞术检测子宫CD45+细胞内细胞因子表达水平.为进一步鉴定CD45+细胞身份,在体外培养系统中采用dsRNA和咪喹莫特刺激胎盘和底蜕膜来源的子官CD3+T细胞和CD49b+NK细胞,并检测细胞内细胞因子表达水平.采用丝裂原激活的蛋白激酶(mitogen-activated protein kinase,MAPK)抑制剂SP600125和PD98059阻断细胞因子表达水平的增加.结果此 dsRNA和咪喹莫特联合刺激对胚胎吸收率的增高具有协同作用,同时对CD45+细胞内TNF-α和IFN-γ的表达增强具有协同刺激作用.进一步细胞鉴定研究显示,虽然在BALB/c小鼠CD3+细胞和CD49b+NK细胞中均可发现这种协同效应,但是在NOD小鼠,这种细胞因子水平的增高应主要归因于CD3+T细胞,因为在CD49b+NK细胞不显示这种细胞因子增高趋势.上述刺激剂合用的协同效应町部分地被JNK(Jun N-terminal kinase)MAPK抑制剂SP600125阻断,而几乎被ERK(extracellular signal.regulated kinase)MAPK抑制剂PD98059所完全阻断.结论增强的TLR3和TLR7联合信号可能是通过NOD小鼠Th1型T细胞,而不是NK细胞所传导.ERK MAPK途径可能在TLR3和TLR7参与的细胞信息传递过程中发挥关键性作用.
목적 연구쌍련RNA(double-stranded RNA,dsRNA)화미규막특연합자격시부구유협동효응.방법 재Toll양수체3(TLR3)적특이성자격제dsRNA、TLR7적특이성자격제미봉막특(R837)단독자격혹이자연합자격조건하,검측BALB/c×C57BL/6화NH세포결함적비비반형당뇨병(NOD)×C57BL/6소서배태흡수솔.채용소서체내주사상술자격제적방법 ,류식세포술검측자궁CD45+세포내세포인자표체수평.위진일보감정CD45+세포신빈,재체외배양계통중채용dsRNA화미규막특자격태반화저세막래원적자관CD3+T세포화CD49b+NK세포,병검측세포내세포인자표체수평.채용사렬원격활적단백격매(mitogen-activated protein kinase,MAPK)억제제SP600125화PD98059조단세포인자표체수평적증가.결과 차 dsRNA화미규막특연합자격대배태흡수솔적증고구유협동작용,동시대CD45+세포내TNF-α화IFN-γ적표체증강구유협동자격작용.진일보세포감정연구현시,수연재BALB/c소서CD3+세포화CD49b+NK세포중균가발현저충협동효응,단시재NOD소서,저충세포인자수평적증고응주요귀인우CD3+T세포,인위재CD49b+NK세포불현시저충세포인자증고추세.상술자격제합용적협동효응정부분지피JNK(Jun N-terminal kinase)MAPK억제제SP600125조단,이궤호피ERK(extracellular signal.regulated kinase)MAPK억제제PD98059소완전조단.결론 증강적TLR3화TLR7연합신호가능시통과NOD소서Th1형T세포,이불시NK세포소전도.ERK MAPK도경가능재TLR3화TLR7삼여적세포신식전체과정중발휘관건성작용.
Objective To investigate the effect of combined double-stranded RNA (dsRNA) and imiquimod stimulation on uterine immune cells. Methods In BALB/c × C57BL/6 mice and non-obese dia-betic (NOD) × C57BI/6 mice, embryo resorption rate was detected in the presence or absence of Toll-like receptor 3 (TLR3) agonist dsRNA [poly( 1: C)], TLR7 agonist imiquimod ( R837), or their combination, respectively. In in vivo system, the status of intracellular cytokine production in uterine CD45 + cells was de-tected by flow cytometry. To identify the CD45 + cells, uterine CD3+ T cells and CD49b + NK cells derived from placenta and decidua basalis were stimulated with dsRNA and imiquimod in in vitro systems, and the status of intracellular cytokine production was detected. Mitogen-activated protein kinase (MAPK) antago- nists SP600125 and PD98059 were used to block the increase of cytokine production. Results A synergistic increase of embryo resorption was observed after the induction of dsRNA and imiquimod combination. Mean-while, a synergistic increase of TNF-α and IFN-γ production was detected after the induction in CD45 + cells. Further study found that although synergistic effect can be detected in both CD3 + cells and CD49b + cells in BALB/c mice, the status was different in NOD mice. The cytokine increase should mainly be attrib-uted to CD3 + T cells, since no such increase was detected among the CD49b + NK cells in the NOD mice. The synergistic effect of combined agonists was partially inhibited by Jun N-terminal kinase (JNK) MAPK inhibitor SP600125 and almost completely abrogated by extracellular signal-regulated kinase (ERK) MAPK inhibitor PD98059. Conclusion Boosted TLR3 and TLR7 signal may be transmitted via Thl-type T cells, rather than NK cells in NOD mice. ERK MAPK pathway may be critical in TLR3 and TLR7 involved signa- ling.