Rieder H, Ramadori G, Meyer zum Büschenfelde K H
I. Medizinische Klinik, Johannes Gutenberg-Universität, Mainz, F.R.G.
J Hepatol. 1988 Dec;7(3):338-44. doi: 10.1016/s0168-8278(88)80006-0.
Guinea pig Kupffer cells were obtained by partial digestion of the liver with pronase and collagenase and purified by centrifugal elutriation. Cells were kept in monolayer culture and their capacity to secrete superoxide anion in response to phagocytosis of zymosan was determined by the cytochrome c method. Compared to resident peritoneal macrophages, Kupffer cells produced somewhat less superoxide (60% +/- 30%). Both cell types were activated by 24 h preincubation with lipopolysaccharide from Salmonella minnesota or muramyl dipeptide to give twice as high a superoxide response to zymosan. The same effect was achieved when Kupffer cells in vitro were incubated for 3 days with supernatants from phytohaemagglutinin-activated peripheral T lymphocytes or recombinant gamma-interferon. These data demonstrate that the resident macrophages of the liver, the Kupffer cells, are able to increase their capacity to secrete reactive oxygen intermediates after proper activation; this fact is possibly important in the pathogenesis of hepatocyte damage upon inflammatory reactions in the liver.
通过用链霉蛋白酶和胶原酶对肝脏进行部分消化来获取豚鼠库普弗细胞，并通过离心淘洗法进行纯化。细胞保持单层培养，采用细胞色素c法测定其在吞噬酵母聚糖后分泌超氧阴离子的能力。与驻留腹膜巨噬细胞相比，库普弗细胞产生的超氧化物略少（60%±30%）。两种细胞类型经用来自明尼苏达沙门氏菌的脂多糖或胞壁酰二肽预孵育24小时后被激活，对酵母聚糖产生的超氧化物反应增加一倍。当体外培养的库普弗细胞与植物血凝素激活的外周T淋巴细胞的上清液或重组γ干扰素孵育3天时，也能获得相同效果。这些数据表明，肝脏的驻留巨噬细胞即库普弗细胞，在适当激活后能够提高其分泌活性氧中间体的能力；这一事实在肝脏炎症反应时肝细胞损伤的发病机制中可能具有重要意义。
