Basista M H, Gavaler J, Stieffenhofer A, Love K, Rosenblum E, Dindzans V J
Division of Gastroenterology, University of Pittsburgh, Pennsylvania.
Alcohol Clin Exp Res. 1993 Jun;17(3):556-60. doi: 10.1111/j.1530-0277.1993.tb00799.x.
Kupffer cells are resident macrophages in the liver and are important in both local and systemic immune responses. We evaluated the ability of Kupffer cells in vitro to respond to immune stimulation after both acute exposure to ethanol and after long-term ethanol consumption of ethanol. Triplets of female Wistar rats were fed a liquid diet containing 0, 12, or 36% ethanol isocalorically for 112 days. When killed, the Kupffer cells were isolated by collagenase perfusion and adhered to plastic 24-well plates. They were then stimulated with 10 micrograms/ml lipopolysaccharide for 4.5 hr. Synthesis of procoagulant activity (PCA) and tumor necrosis factor (TNF), expressions of macrophage response to immune stimuli, were measured by a one-step clotting assay and L929 cytotoxicity assay, respectively. Within each of the 10 triplets, PCA and TNF levels were normalized and expressed as a percentage of the zero ethanol isocaloric control rat. The high ethanol group had significantly lower baseline and stimulated PCA and TNF levels than the low ethanol group. For evaluation of the effect of acute exposure to ethanol, Kupffer cells were stimulated with lipopolysaccharide and varying concentrations (0-400 mg/dl) of ethanol. Cells were incubated for 4.5 hr and assayed for PCA and TNF activity. There was dose-dependent inhibition of PCA and TNF, with increasing concentrations of ethanol. These results indicate that whereas exposure to high levels of ethanol depresses Kupffer cell function, lower levels may be immunostimulatory.
库普弗细胞是肝脏中的常驻巨噬细胞，在局部和全身免疫反应中都很重要。我们评估了库普弗细胞在体外对急性接触乙醇和长期摄入乙醇后免疫刺激的反应能力。将雌性Wistar大鼠三联体等热量地喂食含0%、12%或36%乙醇的液体饮食112天。处死时，通过胶原酶灌注分离库普弗细胞，并使其贴附于塑料24孔板。然后用10微克/毫升脂多糖刺激它们4.5小时。分别通过一步凝血试验和L929细胞毒性试验测量促凝血活性（PCA）和肿瘤坏死因子（TNF）的合成，这是巨噬细胞对免疫刺激反应的表达。在10个三联体中的每一个内，PCA和TNF水平进行归一化处理，并表示为零乙醇等热量对照大鼠的百分比。高乙醇组的基线以及刺激后的PCA和TNF水平显著低于低乙醇组。为了评估急性接触乙醇的影响，用脂多糖和不同浓度（0 - 400毫克/分升）的乙醇刺激库普弗细胞。细胞孵育4.5小时后，检测PCA和TNF活性。随着乙醇浓度的增加，PCA和TNF受到剂量依赖性抑制。这些结果表明，虽然接触高水平乙醇会抑制库普弗细胞功能，但较低水平可能具有免疫刺激作用。
