Priego Teresa, Granado Miriam, Castillero Estibaliz, Martín Ana Isabel, Villanúa M Angeles, López-Calderón Asunción
Departamento de Fisiología, Facultad de Medicina, Universidad Complutense, Madrid 28040, Spain.
J Endocrinol. 2006 Sep;190(3):847-56. doi: 10.1677/joe.1.06938.
We tested whether endotoxin (lipopolysaccharide, LPS) inhibits IGF-I gene expression in hepatocytes and the possible role of Kupffer cells and nitric oxide (NO) in this effect. LPS decreased IGF-I mRNA in hepatocyte cultures and increased the nitrite + nitrate levels in the culture medium. Furthermore, there was a negative correlation between the IGF-I mRNA and the nitrite+nitrate levels. When hepatocytes were cocultured with Kupffer cells, the inhibitory effect of LPS on IGF-I mRNA was higher than in hepatocyte cultures, but the stimulatory effect on nitrite+nitrate was similar in both conditions. The exogenous NO donated by S-nitroso-n-acetyl-d,l-penicillamide also decreased the IGF-I gene expression in hepatocyte cultures. In addition, two specific inducible NO synthase (iNOS) inhibitors, l-N6-(1-iminoethyl)lysine (l-NIL) and aminoguanidine, prevented the effect of LPS on nitrite+nitrate levels and on IGF-I gene expression in hepatocyte cultures. These data indicate that iNOS-derived NO may cause downregulation of IGF-I gene expression in hepatocytes. However, in cocultures, the iNOS inhibitor l-NIL prevented the effect of LPS on nitrite+nitrate levels, but only attenuated the LPS-induced decrease in IGF-I gene expression. We conclude that in hepatocytes, LPS-induced decrease in IGF-I is mainly due to induction of iNOS, whereas in the presence of Kupffer cells LPS inhibits IGF-I through NO release and through other inhibitory pathways.
我们检测了内毒素(脂多糖,LPS)是否抑制肝细胞中IGF-I基因的表达,以及库普弗细胞和一氧化氮(NO)在此效应中可能发挥的作用。LPS降低了肝细胞培养物中IGF-I mRNA的水平,并提高了培养基中亚硝酸盐+硝酸盐的水平。此外,IGF-I mRNA与亚硝酸盐+硝酸盐水平之间呈负相关。当肝细胞与库普弗细胞共培养时,LPS对IGF-I mRNA的抑制作用高于肝细胞培养,但对亚硝酸盐+硝酸盐的刺激作用在两种情况下相似。由S-亚硝基-n-乙酰-d,l-青霉胺提供的外源性NO也降低了肝细胞培养物中IGF-I基因的表达。此外,两种特异性诱导型一氧化氮合酶(iNOS)抑制剂,L-N6-(1-亚氨基乙基)赖氨酸(L-NIL)和氨基胍,可阻止LPS对肝细胞培养物中亚硝酸盐+硝酸盐水平以及IGF-I基因表达的影响。这些数据表明,iNOS衍生的NO可能导致肝细胞中IGF-I基因表达的下调。然而,在共培养中,iNOS抑制剂L-NIL阻止了LPS对亚硝酸盐+硝酸盐水平的影响,但仅减弱了LPS诱导的IGF-I基因表达的降低。我们得出结论,在肝细胞中,LPS诱导的IGF-I降低主要是由于iNOS的诱导,而在存在库普弗细胞的情况下,LPS通过释放NO和其他抑制途径抑制IGF-I。