Thamilselvan S, Khan S R
Department of Pathology, Immunology and Laboratory Medicine, College of Medicine, University of Florida, Gainesville, USA.
J Nephrol. 1998 Mar-Apr;11 Suppl 1:66-9.
Calcium oxalate (CaOx) nephrolithiasis was induced in male Sprague-Dawley rats by administration of 0.75% ethylene glycol. Urinary excretion of lactate dehydrogenase (LDH) was used as a marker of cellular injury. Lipid peroxides (LP), as marker for free radical injury, were measured as malondialdehyde (MDA) in urine and the kidneys. Urinary oxalate (Ox), LDH, LP, CaOx crystals, and renal LP and CaOx crystal deposits were examined on day 0, 5, 30 and 60 of the experiment. There were significant differences between control and experimental rats in all the parameters except LDH which did not show a significant increase after 15 days. Subconfluent cultures of MDCK and LLCPK1 cells were exposed to various concentrations of oxalate and/or 500 fg/ml CaOx crystals. Cell viability was assayed by trypan blue exclusion, cellular injury was determined by measuring LDH in the media, and free radical injury was measured as MDA contents of the cells. On exposure to both Ox and/or CaOx crystals trypan blue exclusion decreased and LDH and MDA increased significantly in both tissue cultures. LLC-PK1 appeared more sensitive. The results indicate that both oxalate and calcium oxalate crystals are injurious to renal epithelial cells in the kidneys as well as in culture.
通过给予0.75%的乙二醇,在雄性Sprague-Dawley大鼠中诱导草酸钙(CaOx)肾结石形成。乳酸脱氢酶(LDH)的尿排泄用作细胞损伤的标志物。脂质过氧化物(LP)作为自由基损伤的标志物,在尿液和肾脏中以丙二醛(MDA)进行测量。在实验的第0、5、30和60天检查尿草酸(Ox)、LDH、LP、CaOx晶体以及肾脏中的LP和CaOx晶体沉积物。除了LDH在15天后未显示出显著增加外,对照组和实验组大鼠在所有参数上均存在显著差异。将MDCK和LLCPK1细胞的亚汇合培养物暴露于不同浓度的草酸盐和/或500 fg/ml的CaOx晶体中。通过台盼蓝排斥法测定细胞活力,通过测量培养基中的LDH来确定细胞损伤,并以细胞的MDA含量来测量自由基损伤。在暴露于Ox和/或CaOx晶体时,两种组织培养物中的台盼蓝排斥率均降低,LDH和MDA显著增加。LLC-PK1似乎更敏感。结果表明,草酸盐和草酸钙晶体对肾脏中的肾上皮细胞以及培养中的细胞均具有损伤作用。