Vom Dahl S, Hallbrucker C, Lang F, Gerok W, Häussinger D
Medizinische Universitätsklinik, Freiburg, Federal Republic of Germany.
Biochem J. 1991 Sep 15;278 ( Pt 3)(Pt 3):771-7. doi: 10.1042/bj2780771.
The effects of insulin and glucagon on liver cell volume and proteolysis were studied in isolated perfused rat liver. The rate of proteolysis was assessed as [3H]leucine release from single-pass-perfused livers from rats which had been prelabelled in vivo by intraperitoneal injection of [3H]leucine. The intracellular water space was determined from the wash-out profiles of simultaneously added [3H]inulin and [14C]urea. In normo-osmotic (305 mosM) control perfusions the intracellular water space was 548 +/- 10 microliters/g wet mass (n = 44) and was increased by 16.5 +/- 2.6% (n = 6), i.e. by 85 +/- 14 microliters/g, after hypoosmotic exposure (225 mosM). Glucagon (0.1 microM) decreased the intracellular water space by 17 +/- 4% (n = 4), whereas insulin (35 nM) increased the intracellular water space by 9.3 +/- 1.4% (n = 15). Also, in isolated rat hepatocyte suspensions insulin (100 nM) caused cell swelling by 10.7 +/- 1.8% (n = 16), which was fully reversed by glucagon. In perfused liver, insulin-induced cell swelling was accompanied by a hepatic net K+ uptake (4.5 +/- 0.2 mumol/g) and an inhibition of proteolysis by 21 +/- 2% (n = 12); further addition of glucagon led to a net K+ release of 3.8 +/- 0.2 mumol/g (n = 7) and fully reversed the insulin effects on both cell volume and proteolysis. Similarly, insulin-induced cell swelling and inhibition of proteolysis were completely antagonized by hyperosmotic (385 mosM) cell shrinkage. Furthermore, cell swelling and inhibition of proteolysis after hypo-osmotic exposure or amino acid addition were reversed by glucagon-induced cell shrinkage. There was a close relationship between the extent of cell swelling and the inhibition of proteolysis, regardless of whether cell volume was modified by insulin, glucagon or aniso-osmotic exposure. The data show that glucagon and insulin are potent modulators of liver cell volume, at least in part by alterations of cellular K+ balance, and that their opposing effects on hepatic proteolysis can largely be explained by opposing effects on cell volume. It is hypothesized that hormone-induced alterations of cell volume may represent an important, not yet recognized, mechanism mediating hormonal effects on metabolism.
在离体灌注的大鼠肝脏中研究了胰岛素和胰高血糖素对肝细胞体积和蛋白水解的影响。蛋白水解速率通过腹腔注射[³H]亮氨酸在体内预先标记的大鼠单次灌注肝脏中[³H]亮氨酸的释放来评估。细胞内水空间由同时添加的[³H]菊粉和[¹⁴C]尿素的洗脱曲线确定。在等渗(305 mosM)对照灌注中,细胞内水空间为548±10微升/克湿重(n = 44),在低渗暴露(225 mosM)后增加了16.5±2.6%(n = 6),即增加了85±14微升/克。胰高血糖素(0.1微摩尔/升)使细胞内水空间降低了17±4%(n = 4),而胰岛素(35纳摩尔/升)使细胞内水空间增加了9.3±1.4%(n = 15)。此外,在离体大鼠肝细胞悬液中,胰岛素(100纳摩尔/升)使细胞肿胀了10.7±1.8%(n = 16),而胰高血糖素可完全逆转这种肿胀。在灌注肝脏中,胰岛素诱导的细胞肿胀伴随着肝脏净K⁺摄取(4.5±0.2微摩尔/克)和蛋白水解抑制21±2%(n = 12);进一步添加胰高血糖素导致净K⁺释放3.8±0.2微摩尔/克(n = 7),并完全逆转了胰岛素对细胞体积和蛋白水解的影响。同样,胰岛素诱导的细胞肿胀和蛋白水解抑制被高渗(385 mosM)细胞收缩完全拮抗。此外,低渗暴露或添加氨基酸后引起的细胞肿胀和蛋白水解抑制被胰高血糖素诱导的细胞收缩所逆转。无论细胞体积是由胰岛素、胰高血糖素还是渗透压变化所改变,细胞肿胀程度与蛋白水解抑制之间都存在密切关系。数据表明,胰高血糖素和胰岛素是肝细胞体积的有效调节因子,至少部分是通过改变细胞K⁺平衡实现的,并且它们对肝脏蛋白水解的相反作用在很大程度上可以通过对细胞体积的相反作用来解释。据推测,激素诱导的细胞体积变化可能代表一种尚未被认识的重要机制,介导激素对代谢的影响。