Warskulat U, Wettstein M, Häussinger D
Medizinische Universitätsklinik, Klinik für Gastroenterologie, Hepatologie und Infektiologie, Heinrich-Heine-Universität, Düsseldorf,Germany.
Biochem J. 1997 Feb 1;321 ( Pt 3)(Pt 3):683-90. doi: 10.1042/bj3210683.
The effects of aniso-osmotic exposure on taurine transport were studied in H4IIE rat hepatoma cells. Hyperosmotic (405 mosmol/l) exposure of H4IIE cells stimulated Na+-dependent taurine uptake and led to an increase in taurine transporter (TAUT) mRNA levels, whereas hypo-osmotic (205 mosmol/l) exposure diminished both taurine uptake and TAUT mRNA levels when compared with normo-osmotic (305 mosmol/l) control incubations. Taurine uptake increased 30-40-fold upon raising the ambient osmolarity from 205 to 405 mosmol/l. When H4IIE cells and perfused livers were preloaded with taurine, hypo-osmotic cell swelling led to a rapid release of taurine from the cells. The taurine efflux, but not taurine uptake, was sensitive to 4,4'-di-isothiocyanatostilbene-2,2'-disulphonic acid (DIDS), suggestive of an involvement of DIDS-sensitive channels in mediating volume-regulatory taurine efflux. Whereas in both H4IIE rat hepatoma cells and primary hepatocytes TAUT mRNA levels were strongly dependent upon ambient osmolarity, mRNAs for other osmolyte transporters, i.e. the betaine transporter BGT-1 and the Na+/myo-inositol transporter SMIT, were not detectable. In line with this, myo-inositol uptake by H4IIE hepatoma cells was low and was not stimulated by hyperosmolarity. However, despite the absence of BGT-1 mRNA, a slight osmosensitive uptake of betaine was observed, but the rate was less than 10% of that of taurine transport. This study identifies a constitutively expressed and osmosensitive TAUT in H4IIE cells and the use of taurine as a main osmolyte, whereas betaine and myo-inositol play little or no role in the osmolyte strategy in these cells. This is in contrast with rat liver macrophages, in which betaine has been shown to be a major osmolyte.
在H4IIE大鼠肝癌细胞中研究了等渗暴露对牛磺酸转运的影响。高渗(405毫渗摩尔/升)暴露H4IIE细胞刺激了依赖钠的牛磺酸摄取,并导致牛磺酸转运体(TAUT)mRNA水平升高,而低渗(205毫渗摩尔/升)暴露与等渗(305毫渗摩尔/升)对照孵育相比,降低了牛磺酸摄取和TAUT mRNA水平。当环境渗透压从205毫渗摩尔/升提高到405毫渗摩尔/升时,牛磺酸摄取增加了30 - 40倍。当H4IIE细胞和灌注肝脏预先加载牛磺酸时,低渗性细胞肿胀导致牛磺酸从细胞中快速释放。牛磺酸外排而非摄取对4,4'-二异硫氰酸芪-2,2'-二磺酸(DIDS)敏感,提示DIDS敏感通道参与介导容量调节性牛磺酸外排。在H4IIE大鼠肝癌细胞和原代肝细胞中,TAUT mRNA水平都强烈依赖于环境渗透压,而其他渗透溶质转运体的mRNA,即甜菜碱转运体BGT-1和钠/肌醇转运体SMIT,则无法检测到。与此一致的是,H4IIE肝癌细胞对肌醇的摄取较低,且不受高渗刺激。然而,尽管不存在BGT-1 mRNA,但仍观察到对甜菜碱有轻微的渗透压敏感性摄取,但其速率不到牛磺酸转运速率的10%。本研究确定了H4IIE细胞中组成性表达且对渗透压敏感的TAUT,以及牛磺酸作为主要渗透溶质的作用,而甜菜碱和肌醇在这些细胞的渗透溶质策略中作用很小或不起作用。这与大鼠肝脏巨噬细胞形成对比,在大鼠肝脏巨噬细胞中,甜菜碱已被证明是主要的渗透溶质。