ANANDAMIDE INHIBITS MACROPHAGE-MEDIATED KILLING OF TUMOR NECROSIS FACTOR-SENSITIVE CELLS
ANANDAMIDE INHIBITS MACROPHAGE-MEDIATED KILLING OF TUMOR NECROSIS FACTOR-SENSITIVE CELLS
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DOI：
10.1016/0024-3205(95)00190-h
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发表时间：
1995-05-05
期刊：
影响因子：
6.1
通讯作者：
MARCIANOCABRAL, F
中科院分区：
文献类型：
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作者：
CABRAL, GA;TONEY, DM;MARCIANOCABRAL, F
Anandamide（花生四烯乙醇酰胺）被证明可以抑制巨噬细胞介导的对肿瘤坏死因子敏感的小鼠L929成纤维细胞的杀伤。扫描电镜（SEM）显示，与痤疮丙酸杆菌（P. acnes）激活的小鼠腹腔巨噬细胞共培养的L929细胞，要么被破坏，要么表面异常，并有许多点状病变。相比之下，L929细胞与Anandamide （20 mg/kg - 80 mg/kg）或外源性大麻素δ -9-四氢大麻酚（THC, 80 mg/kg）的小鼠巨噬细胞共培养，未出现超微结构异常。细胞毒性实验与扫描电镜同时进行，以确定超微结构观察是否与靶细胞杀伤（通过从L929靶细胞释放放射性标记来测量）相关。痤疮假单胞杆菌激活的巨噬细胞诱导[Cr-51]标记的L929细胞特异性释放41%的放射性标记。相比之下，用痤疮假单胞杆菌和20、40或80 mg/kg Anandamide处理的动物巨噬细胞分别表现出38%、25%或28%的放射性标记特异性释放。同样，用痤疮假单胞杆菌和80 mg/kg四氢大麻酚处理的动物巨噬细胞显示出21%的放射性标记特异性释放。体外细胞毒性研究使用放射性标记的L929靶细胞和RAW264.7小鼠巨噬细胞样细胞的条件培养基，可以确定Anandamide发挥其抑制作用的时间间隔。当巨噬细胞在激活前暴露于Anandamide 1小时获得条件培养基时，对靶细胞杀伤的抑制作用最大。相比之下，从活化前预处理24小时- 48小时的RAW264.7细胞中获得的thc处理巨噬细胞条件培养基对靶细胞杀伤的抑制作用最大。这些结果表明，阿南胺和四氢大麻酚对杀伤tnf敏感靶细胞具有相似的抑制作用。然而，这两种物质产生抑制作用的时间间隔不同。
Anandamide (arachidonoylethanolamide) was shown to inhibit macrophage-mediated killing of tumor necrosis factor-sensitive murine L929 fibroblasts. Scanning electron microscopy (SEM) demonstrated that L929 cells, co-cultured with Propionibacterium acnes (P. acnes)-activated peritoneal macrophages from mice treated with vehicle, were either disrupted or had surface abnormalities and numerous punctate lesions. In contrast, L929 cells co-cultured with macrophages from mice receiving P. acnes in concert with Anandamide (20 mg/kg - 80 mg/kg) or the exogenous cannabinoid delta-9-tetrahydrocannabinol (THC; 80 mg/kg) did not exhibit ultrastructural abnormalities. Cytotoxicity assays were performed in parallel with SEM in order to determine whether ultrastructural observations correlated with target cell killing as measured by release of radiolabel from L929 target cells. P. acnes-activated macrophages from vehicle-treated mice elicited 41% specific release of radiolabel from [Cr-51]-labeled L929 cells. In contrast, macrophages from animals treated with P. acnes and with 20, 40, or 80 mg/kg Anandamide exhibited 38%, 25%, or 28% specific release of radiolabel, respectively. Similarly, macrophages from animals treated with P. acnes and with 80 mg/kg THC exhibited 21% specific release of radiolabel. In vitro cytotoxicity studies using radiolabeled L929 target cells and conditioned medium from RAW264.7 murine macrophage-like cells allowed for determination of the time interval over which Anandamide exerted its inhibitory effect. Maximal inhibition of target cell killing occurred when conditioned medium was obtained from macrophages exposed to Anandamide for 1 hr prior to activation. In contrast, conditioned medium from THC-treated macrophages exerted its maximal inhibition of target cell killing when obtained from RAW264.7 cells pretreated for 24hr - 48hr prior to activation. These results indicate that Anandamide and THC exert a similar inhibition of killing of TNF-sensitive target cells. However, the time interval over which these two substances elicit their suppressive effect differs.