CAJ | 학술논문

背景:脂氧素可以抑制炎症细胞、内皮细胞、小鼠脾脏树突状细胞等合成细胞因子,还可以抑制炎性细胞因子对细胞的生物效应.目的:分析脂氧素受体激动剂 BML-111对人巨细胞病毒感染 THP-1源巨噬细胞的免疫调节作用.方法:用人巨细胞病毒 AD169毒株感染 THP-1源巨噬细胞(MOI=0.5),细胞培养后设立对照组、人巨细胞病毒感染组及人巨细胞病毒+BML-111组.在感染后0,1,2,4,12,24,48,72 h 收集各组细胞培养上清液,以 ELISA 检测各组细胞因子的表达水平; RT-PCR 检测各指标 mRNA 的表达强度;Western blot 检测感染4 h 的细胞核内 p65亚基蛋白表达.结果与结论:与对照组相比,其他两组各细胞因子蛋白及 mRNA 表达明显升高(P <0.05).与人巨细胞病毒感染组相比,人巨细胞病毒+BML-111组白细胞介素1β和肿瘤坏死因子α显著降低,转化生长因子β显著升高(P <0.05),白细胞介素10差异无显著性意义(P >0.05);人巨细胞病毒+BML-111组各细胞因子 mRNA 均明显降低(P <0.05).与对照组相比,其他两组细胞核内 NF-κB p65亚基蛋白浓度明显升高(P <0.05);人巨细胞病毒+BML-111组显著低于人巨细胞病毒感染组(P <0.05).说明BML-111可能通过抑制 NF-κB 的 p65亚基核转位,减少白细胞介素1β、肿瘤坏死因子α的表达,促进转化生长因子β的表达,从而对人巨细胞病毒感染的 THP-1源巨噬细胞发挥其免疫负调节作用.
배경:지양소가이억제염증세포、내피세포、소서비장수돌상세포등합성세포인자,환가이억제염성세포인자대세포적생물효응.목적:분석지양소수체격동제 BML-111대인거세포병독감염 THP-1원거서세포적면역조절작용.방법:용인거세포병독 AD169독주감염 THP-1원거서세포(MOI=0.5),세포배양후설립대조조、인거세포병독감염조급인거세포병독+BML-111조.재감염후0,1,2,4,12,24,48,72 h 수집각조세포배양상청액,이 ELISA 검측각조세포인자적표체수평; RT-PCR 검측각지표 mRNA 적표체강도;Western blot 검측감염4 h 적세포핵내 p65아기단백표체.결과여결론:여대조조상비,기타량조각세포인자단백급 mRNA 표체명현승고(P <0.05).여인거세포병독감염조상비,인거세포병독+BML-111조백세포개소1β화종류배사인자α현저강저,전화생장인자β현저승고(P <0.05),백세포개소10차이무현저성의의(P >0.05);인거세포병독+BML-111조각세포인자 mRNA 균명현강저(P <0.05).여대조조상비,기타량조세포핵내 NF-κB p65아기단백농도명현승고(P <0.05);인거세포병독+BML-111조현저저우인거세포병독감염조(P <0.05).설명BML-111가능통과억제 NF-κB 적 p65아기핵전위,감소백세포개소1β、종류배사인자α적표체,촉진전화생장인자β적표체,종이대인거세포병독감염적 THP-1원거서세포발휘기면역부조절작용.
BACKGROUND: Lipoxin can inhibit the synthesis of inflammatory cells, endothelial cells and mouse spleen dendritic cells into the cytokines, and it can also inhibit the biological effects of inflammatory cytokines on cells. OBJECTIVE: To investigate the negative immunomodulatory effect of lipoxin receptor stimulating agent BML-111 on THP-1 macrophages infected by human cytomegalovirus. METHODS: THP-1 derived macrophages were infected with human cytomegalovirus AD169 (multiplicity of infection=0.5), and the cultured cells were randomly divided into control group, human cytomegalovirus group and human cytomegalovirus+BML-111 group. Then the cel culture supernatant was col ected at 0, 1, 2, 4, 12, 24, 48 and 72 hours after infection, and the expression levels of cytokines in each group were detected with enzyme-linked immunosorbent assay; mRNA levels of the factors were tested with real-time PCR; Western blot was used to detect the expression of p65 subunit protein in the nucleus after infection for 4 hours. RESULTS AND CONCLUSION: Compared with the control group, the cytokine protein and mRNA expression both in human cytomegalovirus group and human cytomegalovirus+BML-111 group were increased significantly (P < 0.05). Compared with human cytomegalovirus group, the levels of interleukin-1β and tumor necrosis factor αin human cytomegalovirus+BML-111 group were decreased significantly, and thus the level of transforming growth factor-β was increased greatly (P < 0.05). There was no significant difference of the level of interleukin-10 between the two groups (P > 0.05) mRNA expression. mRNA expression of al the cytokines in human cytomegalovirus+BML-111 group was lower than that in human cytomegalovirus group (P < 0.05). Compared with the control group, the level of p65 subunit protein in the nucleus of human cytomegalovirus group and human cytomegalovirus+BML-111 group was increased significantly (P < 0.05). The level of p65 subunit protein in the nucleus of human cytomegalovirus+BML-111 group was lower than that of human cytomegalovirus group (P <0.05). BML-111 may decrease the expression of interleukin-1β and tumor necrosis factor α, and promote the expression of transforming growth factor-β by inhibiting the nuclear translocation of nuclear factor-κB p65. Thus it plays negative immunoregulation effect on THP-1 macrophages infected by human cytomegalovirus.