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从动物和细胞水平观察螯合剂N,N’-1,2-亚乙基双[N-(2,3-二羟基苯甲基)]甘氨酸(BPCBG)对铀的促排效果,以及对铀致人肾近曲小管上皮细胞(HK-2)损伤的保护作用。大鼠腹腔注射(ip)醋酸铀酰后立即肌内注射(im)不同剂量的BPCBG,采用ICP-MS方法测定24 h的尿铀排出量和肾、骨中铀蓄积量。人肾近曲小管上皮HK-2细胞染铀24 h后给予不同剂量的BPCBG作用24 h,采用ICP-MS方法检测细胞内铀含量。不同剂量醋酸铀酰染毒HK-2细胞后立即或延迟24 h给予BPCBG作用24或48 h,采用CCK-8试剂盒检测细胞存活率,采用CB微核法观察BPCBG对铀致染色体损伤的影响,采用DCFH-DA荧光探针法检测BPCBG对铀诱导细胞内氧自由基生成的影响。以上实验均以DTPA-CaNa3作对照。结果表明,BPCBG(60、120和600μmol.kg-1)使24 h尿铀排出量较铀中毒对照组明显增加约37%～61%,肾、骨中铀蓄积量降低至中毒对照组的41%～31%和86%～42%,促排效果随给药剂量增加而明显增强。HK-2细胞铀染毒后延迟24 h给予10～250μmol.L-1 BPCBG作用24 h,使细胞内铀含量较铀染毒组显著降低约55%～60%,并能明显提高铀染毒HK-2细胞的存活率,显著降低铀诱导的微核形成,有效抑制铀诱导的细胞内氧自由基的产生。DTPA-CaNa3虽能明显降低大鼠肾铀蓄积量和细胞内铀含量,但促排效果显著低于BPCBG,且对铀致细胞损伤无保护作用。以上动物和细胞实验均证明BPCBG是有效的铀促排剂,明显优于DTPA-CaNa3,并具有清除铀诱导细胞内氧自由基产生的作用,可以保护铀致肾细胞损伤,值得进一步研究。
The effects of chelator N, N’-ethylenebis [N- (2,3-dihydroxybenzyl)] glycine (BPCBG) on the uranium emission were observed at animal and cell levels, Protective effect of human renal proximal tubule epithelial cells (HK-2) injury. Rats were injected intraperitoneally (ip) uranyl acetate intramuscularly (im) different doses of BPCBG, measured by ICP-MS method of urinary uranium output and renal, bone uranium volume. Human renal proximal tubule epithelial HK-2 cells were exposed to different doses of BPCBG for 24 h after being exposed to uranium for 24 h. ICP-MS was used to detect intracellular uranium content. The effects of BPCBG on the uranium-induced chromosome damage were observed by CCK-8 kit after exposure to different dosages of uranyl acetate (HK-2) cells exposed to BPCBG for 24 or 48 h immediately or 24 h after exposure to different doses of uranyl acetate , DCFH-DA fluorescent probe was used to detect the effect of BPCBG on the generation of oxygen free radicals in uranium-induced cells. The above experiments were DTPA-CaNa3 as a control. The results showed that BPCBG (60,120 and 600μmol.kg-1) significantly increased uranium excretion by 24 h compared with uranium poisoning control group by 37% -61%, while uranium accumulation in kidney and bone decreased to 41% % ~ 31% and 86% ~ 42%. The effect of promoting blood flow evidently increased with the increase of dose. HK-2 cells were treated with 10 ~ 250μmol.L-1 BPCBG for 24 h after being exposed to uranium for 24 h, which markedly reduced the intracellular uranium content by about 55% -60% and significantly increased the uranium toxicity The survival rate of HK-2 cells significantly reduced uranium-induced micronucleus formation and inhibited uranyl-induced intracellular oxygen free radical production. Although DTPA-CaNa3 could significantly reduce the amount of renal uranium accumulation and intracellular uranium content in rats, the promoting effect of DTPA-CaNa3 was significantly lower than that of BPCBG and had no protective effect on uranium induced cell injury. The above animal and cell experiments have proved that BPCBG is an effective uranium promoting agent, which is obviously superior to DTPA-CaNa3, and has the function of clearing uranium-induced intracellular oxygen free radicals and can protect uranium-induced renal cell injury, which deserves further study.