中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2014年
10期
1742-1747
,共6页
赵莉%姚树桐%陈军%苗成%李严严%田华%周健%翟雷%桑慧%王义围%秦树存
趙莉%姚樹桐%陳軍%苗成%李嚴嚴%田華%週健%翟雷%桑慧%王義圍%秦樹存
조리%요수동%진군%묘성%리엄엄%전화%주건%적뢰%상혜%왕의위%진수존
动脉粥样硬化%载脂蛋白A-I模拟肽%内质网应激%清道夫受体A1%泡沫细胞
動脈粥樣硬化%載脂蛋白A-I模擬肽%內質網應激%清道伕受體A1%泡沫細胞
동맥죽양경화%재지단백A-I모의태%내질망응격%청도부수체A1%포말세포
Atherosclerosis%Apolipoprotein A-I mimetic peptide%Endoplasmic reticulum stress%Scavenger receptor A1%Foam cells
目的：研究载脂蛋白A-I模拟肽D-4F对氧化低密度脂蛋白（ oxidized low-density lipoprotein，ox-LDL）所诱导的巨噬细胞源性泡沫细胞清道夫受体A1（scavenger receptor A1，SR-A1）的抑制作用及其机制。方法：体外培养RAW264．7巨噬细胞，给予不同浓度的D-4F（12．5、25和50 mg/L）、紊乱模拟肽sD-4F（50 mmol/L）处理1 h或者5 mmol/L内质网应激（ endoplasmic reticulum stress，ERS）抑制剂4-苯丁酸处理30 min后，再加入ox-LDL （100 mg/L）继续培养12 h。另外培养巨噬细胞给予50 mg/L D-4F或sD-4F处理1 h，再加入2 mg/L ERS诱导剂衣霉素（ tunicamycin，TM）处理4 h。 MTT法检测细胞活力；试剂盒检测细胞内总胆固醇含量；分别采用免疫印迹法和实时荧光定量聚合酶链反应（ real-time PCR）技术检测SR-A1和ERS标志分子葡萄糖调节蛋白78（ glucose-regula-ted protein 78，GRP78）蛋白和mRNA表达变化；采用多功能酶标仪检测DiI-ox-LDL摄取情况。结果： D-4F明显减轻ox-LDL所诱导的巨噬细胞损伤和细胞内的胆固醇蓄积。 ox-LDL可显著上调SR-A1和GRP78表达，而D-4F对上述变化具有明显抑制作用，且呈浓度依赖性。 D-4F显著抑制TM所诱导的SR-A1和GRP78蛋白水平以及巨噬细胞对ox-LDL的摄取。结论：D-4F可通过抑制SR-A1表达减轻ox-LDL所诱导的巨噬细胞内胆固醇蓄积和细胞损伤，其机制可能与抑制GRP78介导的ERS信号途径有关。
目的：研究載脂蛋白A-I模擬肽D-4F對氧化低密度脂蛋白（ oxidized low-density lipoprotein，ox-LDL）所誘導的巨噬細胞源性泡沫細胞清道伕受體A1（scavenger receptor A1，SR-A1）的抑製作用及其機製。方法：體外培養RAW264．7巨噬細胞，給予不同濃度的D-4F（12．5、25和50 mg/L）、紊亂模擬肽sD-4F（50 mmol/L）處理1 h或者5 mmol/L內質網應激（ endoplasmic reticulum stress，ERS）抑製劑4-苯丁痠處理30 min後，再加入ox-LDL （100 mg/L）繼續培養12 h。另外培養巨噬細胞給予50 mg/L D-4F或sD-4F處理1 h，再加入2 mg/L ERS誘導劑衣黴素（ tunicamycin，TM）處理4 h。 MTT法檢測細胞活力；試劑盒檢測細胞內總膽固醇含量；分彆採用免疫印跡法和實時熒光定量聚閤酶鏈反應（ real-time PCR）技術檢測SR-A1和ERS標誌分子葡萄糖調節蛋白78（ glucose-regula-ted protein 78，GRP78）蛋白和mRNA錶達變化；採用多功能酶標儀檢測DiI-ox-LDL攝取情況。結果： D-4F明顯減輕ox-LDL所誘導的巨噬細胞損傷和細胞內的膽固醇蓄積。 ox-LDL可顯著上調SR-A1和GRP78錶達，而D-4F對上述變化具有明顯抑製作用，且呈濃度依賴性。 D-4F顯著抑製TM所誘導的SR-A1和GRP78蛋白水平以及巨噬細胞對ox-LDL的攝取。結論：D-4F可通過抑製SR-A1錶達減輕ox-LDL所誘導的巨噬細胞內膽固醇蓄積和細胞損傷，其機製可能與抑製GRP78介導的ERS信號途徑有關。
목적：연구재지단백A-I모의태D-4F대양화저밀도지단백（ oxidized low-density lipoprotein，ox-LDL）소유도적거서세포원성포말세포청도부수체A1（scavenger receptor A1，SR-A1）적억제작용급기궤제。방법：체외배양RAW264．7거서세포，급여불동농도적D-4F（12．5、25화50 mg/L）、문란모의태sD-4F（50 mmol/L）처리1 h혹자5 mmol/L내질망응격（ endoplasmic reticulum stress，ERS）억제제4-분정산처리30 min후，재가입ox-LDL （100 mg/L）계속배양12 h。령외배양거서세포급여50 mg/L D-4F혹sD-4F처리1 h，재가입2 mg/L ERS유도제의매소（ tunicamycin，TM）처리4 h。 MTT법검측세포활력；시제합검측세포내총담고순함량；분별채용면역인적법화실시형광정량취합매련반응（ real-time PCR）기술검측SR-A1화ERS표지분자포도당조절단백78（ glucose-regula-ted protein 78，GRP78）단백화mRNA표체변화；채용다공능매표의검측DiI-ox-LDL섭취정황。결과： D-4F명현감경ox-LDL소유도적거서세포손상화세포내적담고순축적。 ox-LDL가현저상조SR-A1화GRP78표체，이D-4F대상술변화구유명현억제작용，차정농도의뢰성。 D-4F현저억제TM소유도적SR-A1화GRP78단백수평이급거서세포대ox-LDL적섭취。결론：D-4F가통과억제SR-A1표체감경ox-LDL소유도적거서세포내담고순축적화세포손상，기궤제가능여억제GRP78개도적ERS신호도경유관。
AIM:To investigate the inhibitory effect of apolipoprotein A-I mimetic peptide D-4F on the scaven-ger receptor A1 ( SR-A1 ) in macrophage-derived foam cells induced by oxidized low-density lipoprotein ( ox-LDL ) . METHODS:RAW264.7 cells were pretreated with different concentrations (12.5, 25 and 50 mg/L) of D-4F or 50 mg/L inactive control peptide scrambled D-4F (sD-4F) for 1 h or endoplasmic reticulum stress (ERS) inhibitor 4-phenylbutyr-ic acid (5 mmol/L) for 30 min, followed by the treatment with 100 mg/L ox-LDL for 12 h.In addition, the cells were pre-treated with 50 mg/L D-4F or sD-4F for 1 h, and then stimulated with 2 mg/L tunicamycin (TM;an ERS inducer), for 4 h.The viability of the cells was measured by MTT assay, and the content of intracellular total cholesterol ( TC) was meas-ured by a tissue/cell TC assay.The protein and mRNA levels of SR-A1 and glucose-regulated protein 78 (GRP78) were analyzed by Western blotting and quantitative real-time PCR, respectively.The fluorescence intensity of DiI-ox-LDL in the cells was detected by a multifunctional microplate reader.RESULTS:D-4F significantly reduced ox-LDL-induced macro-phage injury and intracellular cholesterol accumulation, and attenuated the ox-LDL-induced expression of SRA1 and GRP78 in a dose-dependent manner.Additionally, D-4F significantly inhibited the TM-induced protein expression of SR-A1 and GRP78, and attenuated the uptake of ox-LDL by macrophages.CONCLUSION: D-4F reduces ox-LDL-induced macro-phage cholesterol accumulation and injury by inhibiting SR-A1 expression.The mechanism may be related to the inhibition of ERS signaling pathway mediated by GRP78.
