CAJ | 학술논문

目的探讨氧化低密度脂蛋白模拟的高脂环境对流感病毒感染巨噬细胞的影响并阐明其内在机制.方法分别运用流式细胞术,MTT法检测有无氧化低密度脂蛋白预处理,不同剂量流感病毒感染THP-1巨噬细胞8h、24 h、48 h后,细胞存活率和凋亡率的变化.RT-PCR、Western blot检测内质网应激相关分子GRP78、GADD153的mRNA和蛋白水平.结果 ①流感病毒能感染THP-1巨噬细胞并造成细胞凋亡.②低剂量氧化低密度脂蛋白(＜5μg/mL)促进细胞存活,高剂量氧化低密度脂蛋白(＞ 10 μg/mL)促进细胞凋亡.③5μg/mL氧化低密度脂蛋白单独预处理不影响巨噬细胞凋亡率(P =0.159),但显著增加流感病毒(1×103、2×103、5×103和1×104 TCID50/mL)感染状态下的凋亡率(P =0.001、0.001、0.026和0.035).④氧化低密度脂蛋白增强流感病毒感染引起的内质网相关分子GRP78和GADD153表达,促进巨噬细胞凋亡.结论低剂量氧化低密度脂蛋白预处理可以增加流感病毒感染后巨噬细胞的凋亡,内质网应激在其中起重要作用.
목적 탐토양화저밀도지단백모의적고지배경대류감병독감염거서세포적영향병천명기내재궤제.방법 분별운용류식세포술,MTT법검측유무양화저밀도지단백예처리,불동제량류감병독감염THP-1거서세포8h、24 h、48 h후,세포존활솔화조망솔적변화.RT-PCR、Western blot검측내질망응격상관분자GRP78、GADD153적mRNA화단백수평.결과 ①류감병독능감염THP-1거서세포병조성세포조망.②저제량양화저밀도지단백(＜5μg/mL)촉진세포존활,고제량양화저밀도지단백(＞ 10 μg/mL)촉진세포조망.③5μg/mL양화저밀도지단백단독예처리불영향거서세포조망솔(P =0.159),단현저증가류감병독(1×103、2×103、5×103화1×104 TCID50/mL)감염상태하적조망솔(P =0.001、0.001、0.026화0.035).④양화저밀도지단백증강류감병독감염인기적내질망상관분자GRP78화GADD153표체,촉진거서세포조망.결론 저제량양화저밀도지단백예처리가이증가류감병독감염후거서세포적조망,내질망응격재기중기중요작용.
Objective To investigate the effect and underlying mechanism of influenza virus infection in macrophages especially in low oxidized density lipoprotein (oxLDL) mimicked high lipid conditions.Methods Cell viability and apoptosis were detected by MTT assay and flow cytometry when macrophages were infected by different concentrations of influenza virus for 8 h,24 h and 48 h in the presence or absence of oxLDL.The expressions of GRP78,GADD153 mRNA were detected by RT-PCR and Western blot analysis.Results ①Influenza virus could infect THP-1 macrophages and induce cell apoptosis.②Low level oxLDL(＜ 5 μg/mL)promoted cell survival,in contrast,high level oxLDL (＞ 10 μg/mL)promoted cell apoptosis.③OxLDL pretreatment at 5 μg/mL did not influence macrophage apoptosis sole (P =0.159),on the contrary,it increased macrophages apoptosis along with different concentrations (1 × 103,2 × 103,5 × 103 and 1 × 104TCID50/mL) of influenza virus infection.The P values were 0.001,0.001,0.026 and 0.035,respectively.④The expressions of GRP78 and GADD153 related to endoplasmic reticulum stress were significantly inclined compared with oxLDL absence groups in influenza virus infected conditions.Conclusions Endoplasmic reticulum stress plays a role in oxLDL pretreatment to promote influenza virus infection induced macrophages apoptosis.