Janciauskiene S, Ahrén B
Department of Medicine, Lund University, Malmö, S-20502, Sweden.
Biochem Biophys Res Commun. 1998 Oct 29;251(3):888-93. doi: 10.1006/bbrc.1998.9574.
We studied whether fibrils spontaneously formed by islet amyloid polypeptide (IAPP, also designated amylin) are cytotoxic to insulin producing cells by examining two different insulin producing cell lines, HIT-T15 and RINm5F. IAPP fibrils (</=10microM) added to HIT-T15 cells for one week did not diminish cell viability (tetrazolium bioreduction) or DNA synthesis (3H-thymidine incorporation) nor did it increase cell death (trypan blue staining) or degree of apoptosis (TUNEL assay), and glucose-stimulated insulin secretion and the cytosolic concentration of Ca2+ were unaffected. Similarly, control fibrils (Alzheimer's peptide, Abeta1-42, fibrils) did not reduce cellular function. In contrast, IAPP fibrils decreased cell viability (tetrazolium bioreduction) and increased number of apoptotic cells in RINm5F cells. Furthermore, hydrogen peroxide markedly impaired tetrazolium bioreduction in RINm5F cells but not in HIT-T15 cells. Glutathione reductase activity was increased by IAPP fibrils in RINm5F cells but not in HIT-T15 cells. Our data suggest a different sensitivity for the cytotoxic action of IAPP fibrils between RINm5F and HIT-T15 cells, which may be ascribed to different sensitivity to formation and action of oxygen intermediates.
我们通过检测两种不同的胰岛素生成细胞系HIT-T15和RINm5F,研究胰岛淀粉样多肽(IAPP,也称为胰淀素)自发形成的原纤维对胰岛素生成细胞是否具有细胞毒性。将IAPP原纤维(≤10μM)添加到HIT-T15细胞中一周,并未降低细胞活力(四氮唑生物还原法)或DNA合成(3H-胸腺嘧啶核苷掺入法),也未增加细胞死亡(台盼蓝染色法)或凋亡程度(TUNEL检测法),且葡萄糖刺激的胰岛素分泌和胞质Ca2+浓度均未受影响。同样,对照原纤维(阿尔茨海默病肽,Aβ1-42原纤维)也未降低细胞功能。相比之下,IAPP原纤维降低了RINm5F细胞的活力(四氮唑生物还原法)并增加了凋亡细胞数量。此外,过氧化氢显著损害了RINm5F细胞的四氮唑生物还原能力,但对HIT-T15细胞没有影响。IAPP原纤维使RINm5F细胞中的谷胱甘肽还原酶活性增加,但对HIT-T15细胞没有影响。我们的数据表明,RINm5F细胞和HIT-T15细胞对IAPP原纤维的细胞毒性作用具有不同的敏感性,这可能归因于对氧中间体形成和作用的不同敏感性。