Development of an in vitro assay for the screening of substances capable of dissolving calcium oxalate crystals

被引:11
作者
Saso, L [1 ]
Valentini, G [1 ]
Leone, MG [1 ]
Grippa, E [1 ]
Silvestrini, B [1 ]
机构
[1] Univ Rome La Sapienza, Inst Pharmacol & Pharmacognosy, I-00185 Rome, Italy
关键词
calcium oxalate dissolution; chemolysis; renal stones; nephrolithiasis;
D O I
10.1159/000030331
中图分类号
R5 [内科学]; R69 [泌尿科学(泌尿生殖系疾病)];
学科分类号
1002 ; 100201 ;
摘要
Despite the risk of kidney damage, lithotripsy is the usual way of treating calcium oxalate (CaOx) stones, the most common type of nephrolithiasis, because no effective chemolytic agents are available. However, the search of new calcium chelators, less toxic than the current ones, continues, and some of them could be tested in experimental models of nephrolithiasis, after their ability of dissolving CaOx crystals is verified. In this connection, we developed a simple assay that requires only inexpensive equipment available in most laboratories for the screening of substances potentially capable of dissolving CaOx crystals. In particular, we decided to investigate whether substances previously shown to inhibit CaOx precipitation were also capable of dissolving this salt. Briefly, CaOx tablets of highly reproducible weight (4.55 +/- 0.07 mg) were prepared by spinning, at high speed (16,000 g), microcentrifuge tubes in which 500 mu l aliquots of 0.1 M sodium oxalate and 0.1 M calcium chloride at pH 6 were added. When these tablets were incubated overnight with solutions at different concentrations of EDTA, sodium citrate, manganese chloride, sodium sulfate, sodium chloride, malic acid, succinic acid and gluconic acid, a significant dissolving activity was observed for EDTA (similar to 25% at 0.25 M), sodium citrate (similar to 30% at 1 M) and manganese chloride (similar to 20% at 0.5 M). A good linear correlation (r(2) = 0.84, p < 0.05) was Found between the affinity for calcium and the activity of EDTA, sodium citrate, sodium sulfate, malic acid, succinic acid and gluconic acid, indicating that these compounds act mainly by chelating the calcium ion. Instead, manganese was supposed to act by interacting with the oxalate ion.
引用
收藏
页码:210 / 214
页数:5
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