Geier Andreas, Kim Suk Kyum, Gerloff Thomas, Dietrich Christoph G, Lammert Frank, Karpen Saul J, Stieger Bruno, Meier Peter J, Matern Siegfried, Gartung Carsten
Department of Internal Medicine III, University of Technology, Pauwelsstrasse 30, 52074 Aachen, Germany.
J Hepatol. 2002 Aug;37(2):198-205. doi: 10.1016/s0168-8278(02)00108-3.
BACKGROUND/AIMS: Hepatobiliary transporters are down-regulated in cholestasis, but their expression in acute, non-cholestatic, cytokine-mediated liver injury is unknown. Thus we studied the molecular mechanisms, by which sodium taurocholate cotransporting polypeptide (Ntcp), organic anion transporting polypeptide 1 (Oatp1), Oatp2, Oatp4, multidrug-resistance protein 2 (Mrp2) and bile salt export pump (Bsep) are regulated in liver injury induced by carbon tetrachloride (CCl(4)).
mRNA and protein levels were determined in rats 24 and 72h after CCl(4) injection. Transporter gene transcription and binding activities of Ntcp and Mrp2 transactivators were assessed by nuclear runoff and electrophoretic mobility shift assays.
mRNA levels significantly declined to 41+/-44% for Ntcp, 65+/-41% for Oatp1 and 64+/-28% for Oatp2, but remained unchanged for Oatp4, canalicular Mrp2 and Bsep. Protein levels declined only for Oatp4 (-50+/-17%) and Ntcp (-23+/-13%) at 24h. Reduced mRNA levels (Ntcp, Oatp1, Oatp2) were associated with decreased transcriptional activities. Binding activity of Ntcp transactivators (hepatocyte nuclear factor 1 alpha (HNF1alpha) and CAAT enhancer binding protein alpha (C/EBPalpha) were reduced by 24h, whereas retinoid X receptor alpha (RXRalpha):retinoid acid receptor alpha (RARalpha) as transactivator of both Ntcp and Mrp2 remained unaltered. Recovery of acute hepatitis and changes in gene expression occurred after 72h.
Acute liver injury results in down-regulation of basolateral organic anion transporters similar to liver regeneration after partial hepatectomy, but in contrast to endotoxin-induced cholestasis. Maintained binding activity of RXRalpha:RARalpha may explain differences in Mrp2 expression.
背景/目的:肝胆转运体在胆汁淤积时表达下调,但其在急性、非胆汁淤积性、细胞因子介导的肝损伤中的表达尚不清楚。因此,我们研究了在四氯化碳(CCl₄)诱导的肝损伤中,牛磺胆酸钠共转运多肽(Ntcp)、有机阴离子转运多肽1(Oatp1)、Oatp2、Oatp4、多药耐药蛋白2(Mrp2)和胆盐输出泵(Bsep)的调控分子机制。
在注射CCl₄后24小时和72小时测定大鼠的mRNA和蛋白水平。通过核转录延伸试验和电泳迁移率变动分析评估转运体基因转录以及Ntcp和Mrp2反式激活因子的结合活性。
Ntcp的mRNA水平显著下降至41±44%,Oatp1为65±41%,Oatp2为64±28%,但Oatp4、胆小管Mrp2和Bsep的mRNA水平保持不变。蛋白水平仅在24小时时Oatp4(-50±17%)和Ntcp(-23±13%)下降。mRNA水平降低(Ntcp、Oatp1、Oatp2)与转录活性降低相关。Ntcp反式激活因子(肝细胞核因子1α(HNF1α)和CAAT增强子结合蛋白α(C/EBPα))的结合活性在24小时时降低,而作为Ntcp和Mrp2反式激活因子的视黄酸X受体α(RXRα):视黄酸受体α(RARα)保持不变。急性肝炎恢复及基因表达变化在72小时后出现。
急性肝损伤导致基底外侧有机阴离子转运体下调,这与部分肝切除术后的肝再生相似,但与内毒素诱导的胆汁淤积相反。RXRα:RARα结合活性的维持可能解释了Mrp2表达的差异。