摘要
目的探讨亚油酸对人成纤维细胞增殖及其分泌炎症介质的影响及其可能的作用机制。方法体外培养人成纤维细胞,分为正常对照组(不作任何处理)和实验组(细胞培养液中分别加入4mmol/L、2mmol/L、1mmol/L、0.5mmol/L和0.25mmol/L的亚油酸)。不同培养时相点观察细胞的光镜和电镜形态学、台盼蓝染色法绘制生长曲线、MTT法检测增殖抑制、流式细胞仪检测生长周期,ELISA法检测培养上清液中TNFα、IL-1β、IL-6、IL-8的含量,改良Griess法测定NO含量。结果亚油酸培养48h后,细胞增殖明显降低(P<0.05),光镜下见细胞生长缓慢,形态改变;电镜下见胞内脂滴、胞核和线粒体肿胀,部分细胞胞膜与核膜溶解破裂;流式细胞分析显示,G0/G1期、G2/M期细胞增多,S期细胞明显减少。亚油酸浓度为1mmol/L和2mmol/L时,实验组细胞TNFα分泌量显著高于对照组(P<0.05);亚油酸浓度为4mmol/L时,IL-1β、IL-6的分泌量与对照组相比明显升高(P<0.01);亚油酸浓度为2mmol/L和4mmol/L时,细胞IL-8、NO分泌量与对照组相比明显升高(P<0.05)。结论高浓度亚油酸抑制体外培养人成纤维细胞生长、增殖,并促进人成纤维细胞分泌炎症介质。
Objective To investigate the effect of linoleic acid on the proliferation and secreting inflammatory mediatos of cultured fibroblast and mechanisms. Methods Human fibroblasts were cultured in vitro and randomly divided into A （normal control） and B （with treatment of 4 mmol/L,2 mmol/L,1 mmol/L,0.5 mmol/L and 0.25 mmol/L linoleic acid） groups. Flow cytometry analysis was used to detect cell cycle. The levels of TNF-α, IL-1β, different time-points, the dramatic changes in the morphology of cells were observed under light and electron microscope respectively. The levels of IL-6, IL-8 and NO in the supernatants of cultured fibroblast were assayed by enzyme-linked immunosorbent assay （ELISA）and Griess assay. Results From the proliferation curve, it showed the keratinocytes grew markedly, but lowered after 2 days of incubation with linoleic acid（P0.05）. After 48 hours of exposure to linoleic acid, by light microscopy,fibroblasts showed poor growth and morphologic changes; by electron microscopy, many lipid droplet, nuclear and mitochondrion swelling, cellular membrane and nuclear membrane disruption occurred in the treatment fibroblast. The flow cytometry analysis of the cell cycle distribution also revealed that the percentage of phase G0/G1 and G2/M were significantly increased, the percentage of phase S was markedly decreased. The levels of TNFα in the supernatants of cultured fibroblast of 1 mmol/L, 2 mmol/L linoleic acid groups were significantly higher than the control group （P0.05）. The levels of IL-1β,IL-6 in the supernatants of cultured fibroblast of 4 mmol/L linoleic acids groups were significantly higher than the control group （P0.01）. The levels of IL-8, NO in the supernatants of cultured fibroblast of linoleic acids 2 mmol/L,4 mmol/L groups were significantly higher than the control group（P0.05）.Conclusion Hight concentration linoleic acids can inhibit the proliferation and the secretion of inflammatory mediators in cultured fibroblasts.
出处
《中华损伤与修复杂志（电子版）》
CAS
2011年第5期22-25,共4页
Chinese Journal of Injury Repair and Wound Healing(Electronic Edition)
关键词
亚油酸
人成纤维细胞
细胞增殖
炎症
一氧化氮
Linoleic acid
Fibroblast
Cell proliferation
Inflammatory
Nitric oxide
