CAJ | 학술논문

目的：观察姜黄素(curcumin,Cur)对内毒素(lipopolysaccharide,LPS)诱导的血管平滑肌细胞(vascular smooth muscle cells,VSMCs)Toll 样受体4(Toll-like receptor4,TLR4)/还原型烟酰胺腺嘌呤二核苷酸磷酸(nicotinamide adenine dinucleotide phosphate,NADPH)氧化酶/活性氧(reactive oxygen species,ROS)信号通路及炎症因子释放的影响。方法原代培养 VSMCs 分为5组：对照组、LPS 组、LPS+姜黄素5μmol/L 组、LPS+姜黄素10μmol/L 组、LPS+姜黄素30μmol/L 组和姜黄素30μmol/L 组。MTT 法测定不同浓度姜黄素对 VSMCs 细胞活性的影响；酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)测定各组细胞上清液中肿瘤坏死因子-α(tumor necrosis factor-alpha,TNF-α)和白细胞介素-1(interleukin-1,IL-1)蛋白的表达；Real-time PCR 法及 Wester-blot 检测各组细胞 TLR4、p22phox 蛋白的表达；流式细胞仪测定细胞内 ROS 的表达。结果姜黄素在0~80μmol/L 浓度范围内对细胞活性无显著影响。姜黄素(5、10、30μmol/L)可以有效地抑制 LPS 诱导的 TNF-α和 IL-1过分泌和 TLR4、p22phox、ROS 的高表达并具有浓度依赖性。结论姜黄素能够有效地抑制 LPS 诱导的 VSMCs 炎症因子分泌、TLR4表达及其下游 NADPH 氧化酶、ROS 的生成,姜黄素对 LPS 诱导的 VSMCs 炎症反应的抑制作用可能部分依赖于 TLR4介导的 NADPH 氧化酶/ROS 信号通路。
목적：관찰강황소(curcumin,Cur)대내독소(lipopolysaccharide,LPS)유도적혈관평활기세포(vascular smooth muscle cells,VSMCs)Toll 양수체4(Toll-like receptor4,TLR4)/환원형연선알선표령이핵감산린산(nicotinamide adenine dinucleotide phosphate,NADPH)양화매/활성양(reactive oxygen species,ROS)신호통로급염증인자석방적영향。방법원대배양 VSMCs 분위5조：대조조、LPS 조、LPS+강황소5μmol/L 조、LPS+강황소10μmol/L 조、LPS+강황소30μmol/L 조화강황소30μmol/L 조。MTT 법측정불동농도강황소대 VSMCs 세포활성적영향；매련면역흡부법(enzyme linked immunosorbent assay,ELISA)측정각조세포상청액중종류배사인자-α(tumor necrosis factor-alpha,TNF-α)화백세포개소-1(interleukin-1,IL-1)단백적표체；Real-time PCR 법급 Wester-blot 검측각조세포 TLR4、p22phox 단백적표체；류식세포의측정세포내 ROS 적표체。결과강황소재0~80μmol/L 농도범위내대세포활성무현저영향。강황소(5、10、30μmol/L)가이유효지억제 LPS 유도적 TNF-α화 IL-1과분비화 TLR4、p22phox、ROS 적고표체병구유농도의뢰성。결론강황소능구유효지억제 LPS 유도적 VSMCs 염증인자분비、TLR4표체급기하유 NADPH 양화매、ROS 적생성,강황소대 LPS 유도적 VSMCs 염증반응적억제작용가능부분의뢰우 TLR4개도적 NADPH 양화매/ROS 신호통로。
Objective To explore the inhibitory effect of curcumin on LPS-induced inflammation and the activation of Toll-like receptor 4 (TLR4 )/NADPH oxidase/reactive oxygen species (ROS)signaling pathway in vascular smooth muscle cells (VSMCs).Methods Primary VSMCs were cultured and divided into control group, LPS group,LPS + curcumin 5 μmol/L group,LPS + curcumin 10 μmol/L group and LPS + curcumin 30 μmol/L group.Cell activity was observed by MTT assay.The secretion of tumor necrosis factor-α(TNF-α)and interleukin-1 (IL-1)was measured by enzyme linked immunosorbent assay (ELISA)kits.The mRNA expressions of TLR4 and p22phox were detected by real-time PCR.Expression of intracellular ROS was measured by flow cytometry. Results The activities of VSMCs were not significantly affected by curcumin at the concentration between 0 and 80 μmol/L.Curcumin (5,10 and 30 μmol/L)significantly inhibited LPS-induced oversecretion of TNF-αand IL-1, as well as overexpression of TLR4 and p22phox at the mRNA and protein levels,and ROS production in VSMCs in a concentration-dependent manner.Conclusion Curcumin has a concentration-dependent inhibitory effect on the secretion of inflammatory cytokine,overexpressions of TLR4 and p22phox,and production of ROS in VSMCs stimulated by LPS.Furthermore,curcumin may partly depend on TLR4/NADPH oxidase/ROS signaling pathways to inhibit inflammation in LPS-induced VSMCs.