中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2014年
2期
132-135
,共4页
李全忠%翟振丽%马维红%钱宗杰
李全忠%翟振麗%馬維紅%錢宗傑
리전충%적진려%마유홍%전종걸
动脉粥样硬化%载脂蛋白A-Ⅰ%巨噬细胞%炎症
動脈粥樣硬化%載脂蛋白A-Ⅰ%巨噬細胞%炎癥
동맥죽양경화%재지단백A-Ⅰ%거서세포%염증
Atherosclerosis%Apolipoprotein A-Ⅰ%Macrophages%Inflammation
目的 观察载脂蛋白(apo)AⅠ对炎症型巨噬细胞极性的影响,探讨高密度脂蛋白(HDL)抗炎的细胞和分子机制.方法 选取C57BL/6小鼠,培养其骨髓来源巨噬细胞.实验分为3组:(1)对照组:常规培养小鼠骨髓源性巨噬细胞48 h,不给予任何药物处理；(2)模型组:细胞培养24 h后加γ干扰素(100 U/ml)和脂多糖(5 ng/ml)后继续培养24 h;(3)处理组:先予10μg/ml浓度的人apoA Ⅰ孵育细胞24 h,换液后加入γ干扰素(100 U/ml)和脂多糖(5 ng/ml)孵育24 h.应用流式细胞仪检测膜分子CD16/32、CD206的表达；应用ELISA检测白细胞介素(IL)-10和IL-12的分泌；应用实时荧光定量PCR检测Toll样受体4(TLR4)、髓样分化因子88 (MyD88)、干扰素调节因子5(IRF5) mRNA的表达.结果 处理组巨噬细胞CD16/32表达阳性率(91.17％±1.99％比50.47％±1.02％,P＜0.05)、IL-12分泌[(747.27±3.74) pg/ml比(73.80±4.56) pg/ml,P＜0.05]明显低于模型组；CD206阳性率(0.33％±0.12％比3.00％±0.36％,P＜0.05)、IL-10分泌[(23.56±4.30) pg/ml比(32.91±2.47) pg/ml,P＜0.05]明显高于模型组.TLR4 mRNA(1.000±0.025比0.708±0.003,P＜0.05)、MyD88 mRNA(1.591±0.005比1.341±0.005,P＜0.05)和IRF5 mRNA(0.954±0.005比0.463±0.003,P＜0.05)表达均显著低于模型组.结论 apoAⅠ促进炎症型巨噬细胞向抗炎型巨噬细胞转变,此效应可能与抑制TLR4-MyD88-IRF5通路有关.
目的 觀察載脂蛋白(apo)AⅠ對炎癥型巨噬細胞極性的影響,探討高密度脂蛋白(HDL)抗炎的細胞和分子機製.方法 選取C57BL/6小鼠,培養其骨髓來源巨噬細胞.實驗分為3組:(1)對照組:常規培養小鼠骨髓源性巨噬細胞48 h,不給予任何藥物處理；(2)模型組:細胞培養24 h後加γ榦擾素(100 U/ml)和脂多糖(5 ng/ml)後繼續培養24 h;(3)處理組:先予10μg/ml濃度的人apoA Ⅰ孵育細胞24 h,換液後加入γ榦擾素(100 U/ml)和脂多糖(5 ng/ml)孵育24 h.應用流式細胞儀檢測膜分子CD16/32、CD206的錶達；應用ELISA檢測白細胞介素(IL)-10和IL-12的分泌；應用實時熒光定量PCR檢測Toll樣受體4(TLR4)、髓樣分化因子88 (MyD88)、榦擾素調節因子5(IRF5) mRNA的錶達.結果 處理組巨噬細胞CD16/32錶達暘性率(91.17％±1.99％比50.47％±1.02％,P＜0.05)、IL-12分泌[(747.27±3.74) pg/ml比(73.80±4.56) pg/ml,P＜0.05]明顯低于模型組；CD206暘性率(0.33％±0.12％比3.00％±0.36％,P＜0.05)、IL-10分泌[(23.56±4.30) pg/ml比(32.91±2.47) pg/ml,P＜0.05]明顯高于模型組.TLR4 mRNA(1.000±0.025比0.708±0.003,P＜0.05)、MyD88 mRNA(1.591±0.005比1.341±0.005,P＜0.05)和IRF5 mRNA(0.954±0.005比0.463±0.003,P＜0.05)錶達均顯著低于模型組.結論 apoAⅠ促進炎癥型巨噬細胞嚮抗炎型巨噬細胞轉變,此效應可能與抑製TLR4-MyD88-IRF5通路有關.
목적 관찰재지단백(apo)AⅠ대염증형거서세포겁성적영향,탐토고밀도지단백(HDL)항염적세포화분자궤제.방법 선취C57BL/6소서,배양기골수래원거서세포.실험분위3조:(1)대조조:상규배양소서골수원성거서세포48 h,불급여임하약물처리；(2)모형조:세포배양24 h후가γ간우소(100 U/ml)화지다당(5 ng/ml)후계속배양24 h;(3)처리조:선여10μg/ml농도적인apoA Ⅰ부육세포24 h,환액후가입γ간우소(100 U/ml)화지다당(5 ng/ml)부육24 h.응용류식세포의검측막분자CD16/32、CD206적표체；응용ELISA검측백세포개소(IL)-10화IL-12적분비；응용실시형광정량PCR검측Toll양수체4(TLR4)、수양분화인자88 (MyD88)、간우소조절인자5(IRF5) mRNA적표체.결과 처리조거서세포CD16/32표체양성솔(91.17％±1.99％비50.47％±1.02％,P＜0.05)、IL-12분비[(747.27±3.74) pg/ml비(73.80±4.56) pg/ml,P＜0.05]명현저우모형조；CD206양성솔(0.33％±0.12％비3.00％±0.36％,P＜0.05)、IL-10분비[(23.56±4.30) pg/ml비(32.91±2.47) pg/ml,P＜0.05]명현고우모형조.TLR4 mRNA(1.000±0.025비0.708±0.003,P＜0.05)、MyD88 mRNA(1.591±0.005비1.341±0.005,P＜0.05)화IRF5 mRNA(0.954±0.005비0.463±0.003,P＜0.05)표체균현저저우모형조.결론 apoAⅠ촉진염증형거서세포향항염형거서세포전변,차효응가능여억제TLR4-MyD88-IRF5통로유관.
Objective To explore the anti-inflammatory mechanisms of high density lipoprotein (HDL) by observing the effects of apoprotein (apo) A Ⅰ,a major protein component of HDL,on the inflammatory macrophage cell polarity.Methods Cultured mice marrow-derived macrophages were stimulated with lipopolysaccharide and interferon after 10 μg/ml of apoA Ⅰ were added to the macrophages for 24 hours.The expression of membrane molecules CD16/32,CD206 were detected by fluorescence activated cell sorting (FACS).ELISA was used to detect the secretion of IL-10 and IL-12.Real-time quantitative PCR was used to detect the mRNA expression of TLR4,MyD88 and IRF5.Results Compared to macrophages stimulated by interferon and lipopolysaccharide but without pretreatment with apoA Ⅰ,pre-incubation with apoA Ⅰ significantly downregulated the expression of CD16/32 (91.17％ ± 1.99％ vs.50.47％ ± 1.02％,P ＜0.05),IL-12 [(747.27 ±3.74) pg/ml vs.(73.80 ±4.56) pg/ml,P ＜0.05],upregulated the expression of CD206 (0.33％ ± 0.12％ vs.3.00％ ± 0.36％,P ＜ 0.05),IL-10 expression [(23.56 ±4.30) pg/ml vs.(32.91 ± 2.47) pg/ml,P ＜ 0.05],and reduced the mRNA expression of TLR4(1.000±0.025 vs.0.708 ±0.003,P＜0.05),MyD88(1.591 ±0.005 vs.1.341 ±0.005,P＜ 0.05),IRF5 (0.954 ± 0.005 vs.0.463 ± 0.003,P ＜ 0.05).Conclusion ApoA Ⅰ enhances the switch of inflammatory macrophages to anti-inflammatory macrophages possibly through inhibiting TLR4-MyD88-IRF5 pathway.
