Xu Li-Ying, Mu Min, Wang Man-Li, Liu Jin-Cheng, Zhou Yuan-Jie, Wu Jing, Jiang Bing-You, Chen Ming-Gong, Hu Dong, Tao Xing-Rong
School of Medicine, Department of Medical Frontier Experimental Center, Anhui University of Science and Technology, 168 Taifeng Road, Huainan City, Anhui Province 232001, China.
Key Laboratory of Industrial Dust Control and Occupational Health of the Ministry of Education of Anhui Higher Education Institutes, 168 Taifeng Road, Huainan City, Anhui Province 232001, China.
J Clin Biochem Nutr. 2020 Sep;67(2):146-152. doi: 10.3164/jcbn.19-95. Epub 2020 Apr 17.
Our study was to understand the autophagy induce by different ratios and concentrations of LA/DHA on Raw264.7 cell, and then to investigate the effect of Raw264.7 autophagy on the clearance of . Raw264.7 cells was treated by LA/DHA in different concentrations (50/100 µmol/L) and ratios (4:1, 6:1, 8:1, 1:4, 1:6 and 1:8) for 6/12/24 h, cell viability assay was assessed by Cell Counting Kit-8, LC3B, p62, P-mTOR, P-Akt, P-PI3K and BECN 1 were detected by the Western blot. LA/DHA could induce autophagy of Raw264.7 cells through the PI3K-Akt-mTOR signaling pathway, the strong effect on autophagy by the concentration is 100 µmol/L, the ratio is 6:1 of LA/DHA, and the treatment time is 24 h. Compared with the images in the control group obtained by merging red and green fluorescence channels, the treatment of LA, DHA in a ratio of 6:1 at a concentration of 100 µmol/L for 24 h significantly lead to a substantial number of autophagosomes (yellow) as well as autolysosomes (red), enhancing autophagy flux. Autophagy induce by LA/DHA can devour and damage intracellular and extracellular . These results indicate that LA/DHA cloud induce autophagy and enhance the phagocytosis and killing ability of macrophages to intracellular parasitic bacteria.
我们的研究旨在了解不同比例和浓度的LA/DHA对Raw264.7细胞自噬的诱导作用,进而研究Raw264.7细胞自噬对……清除的影响。将Raw264.7细胞用不同浓度(50/100 μmol/L)和比例(4:1、6:1、8:1、1:4、1:6和1:8)的LA/DHA处理6/12/24小时,通过细胞计数试剂盒-8评估细胞活力,用蛋白质免疫印迹法检测LC3B、p62、磷酸化mTOR(P-mTOR)、磷酸化Akt(P-Akt)、磷酸化磷脂酰肌醇-3激酶(P-PI3K)和Beclin 1(BECN 1)。LA/DHA可通过PI3K-Akt-mTOR信号通路诱导Raw264.7细胞自噬,对自噬作用最强的浓度为100 μmol/L,LA/DHA比例为6:1,处理时间为24小时。与通过合并红色和绿色荧光通道获得的对照组图像相比,100 μmol/L浓度、6:1比例的LA和DHA处理24小时可显著导致大量自噬体(黄色)以及自溶酶体(红色),增强自噬通量。LA/DHA诱导的自噬可吞噬和破坏细胞内和细胞外的……。这些结果表明,LA/DHA可诱导自噬并增强巨噬细胞对细胞内寄生菌的吞噬和杀伤能力。