Gentamicin-induced alterations in pig kidney epithelial (LLC-PK1) cells in culture.
Gentamicin-induced alterations in pig kidney epithelial (LLC-PK1) cells in culture.
复制标题
庆大霉素诱导培养的猪肾上皮 (LLC-PK1) 细胞的变化。
DOI：
--
复制
发表时间：
1986
影响因子：
3.5
通讯作者：
M. Venkatachalam
中科院分区：
文献类型：
--
作者：
D. Schwertz;J. Kreisberg;M. Venkatachalam
观察庆大霉素对体外培养的LLC-PK1细胞的影响。在没有抗生素和抗真菌药物的情况下，将庆大霉素(0.5-2.0 mM)加入到培养至融合的细胞的培养液中。暴露于庆大霉素(1-4天)不影响细胞总蛋白或DNA水平、细胞总数或各种标志酶的释放。庆大霉素处理的细胞的ATP水平与对照细胞没有差别；然而，庆大霉素处理的细胞的培养液中乳酸水平明显较低。电子显微镜下的形态检查显示庆大霉素诱导的髓样小体形成。此外，庆大霉素处理的细胞中总磷脂水平显著升高。对特定磷脂类别的分析表明，只有磷脂酰胆碱、磷脂酰肌醇和多磷肌醇磷脂水平随着时间的推移而增加。磷脂酰肌醇的增加百分比最高。将正常中钙浓度(0.2 mg/ml)提高1.5倍、2.0倍或3.0倍不改变庆大霉素诱导的细胞内磷脂酰肌醇和磷脂酰胆碱的升高。庆大霉素暴露还导致LLC-PK1细胞游离脂肪酸、单甘酯、二甘酯和非酯化胆固醇的周转率呈浓度依赖性增加，而甘油三酯的周转率降低。庆大霉素显著抑制钙离子进入和通过细胞单层，尽管45Ca+与庆大霉素处理的细胞结合更强。这些结果表明，庆大霉素在LLC-PK1细胞中的毒性表现与在整个动物中的报道相似，从而使培养的LLC-PK1细胞成为阐明庆大霉素引起肾上皮细胞损伤机制的有效系统。
The effect of gentamicin exposure was investigated in LLC-PK1 cells in culture. Gentamicin (0.5-2.0 mM) was added to the medium of cells which had been grown to confluency in the absence of antibiotics and antimycotics. Exposure to gentamicin (1-4 days) did not effect total cellular protein or DNA levels, total cell number or the release of various marker enzymes to the medium. ATP levels in gentamicin-treated cells did not differ from control cells; however, medium from the gentamicin-treated cells contained significantly lower lactic acid levels. Morphological examination by electron microscopy revealed gentamicin-elicited myeloid body formation. Furthermore, total phospholipid level was elevated markedly in gentamicin-treated cells. Analysis of specific phospholipid classes showed only phosphatidylcholine, phosphatidylinositol and polyphosphoinositide phospholipid levels increased in a time-dependent manner. Phosphatidylinositol showed the highest percentage of increase. Raising the normal medium calcium concentration (0.2 mg/ml) 1.5-, 2.0- or 3.0-fold did not alter gentamicin-induced elevation in cellular phosphatidylinositol and phosphatidylcholine. Gentamicin exposure also resulted in a concentration-dependent increase in the turnover of LLC-PK1 cell-free fatty acids, monoglyceride, diglyceride and nonesterified cholesterol and a decrease in triglyceride turnover. Calcium transport into and through the cell monolayer was inhibited markedly by gentamicin despite the fact that 45Ca++ binding to gentamicin-treated cells was greater. These results demonstrate that manifestations of gentamicin toxicity in LLC-PK1 cells parallel those reported in the whole animal thus making the LLC-PK1 cell in culture a valid system for elucidating the mechanism of gentamicin-elicited alterations in renal epithelium.