摘要
目的比较高渗造影剂(HOCM,泛影葡胺)和低渗造影剂(LOCM,欧乃派克)对人肾小管上皮细胞的毒性,探讨Bax／Bcl-2,半胱氨酸天冬氨酸蛋白酶(caspase)-3在造影剂诱导肾小管上皮细胞凋亡中的作用。方法以HKC细胞株为研究对象．实验分为7组:HOCM1组 (111 mgI／ml),HOCM2组(74 mgI／ml),LOCM1组(111 mgI／ml),LOCM2组(74 mgI／ml),甘露醇高渗对照组,甘露醇低渗对照组,培养基对照组。采用四甲基偶氮唑蓝(MTY)试验和乳酸脱氢酶 (LDH)检测造影剂对体外培养HKC细胞的毒性作用。采用Hoechst染色、TUNEL染色、DNA琼脂糖凝胶电泳、电镜和流式细胞仪DNA分析方法观察造影剂对HKC细胞凋亡的作用。采用 Western印迹方法检测Bax和Bcl-2蛋白含量的变化。采用荧光比色法检测caspase-3活性。结果 HOCM和LOCM组培养液中LDH水平较对照组显著增高(P<0．05),细胞活力较对照组显著降低(P<0．05),且与渗透浓度,作用时间及碘离子浓度有关。HOCM可诱导肾小管上皮细胞凋亡,且明显高于甘露醇高渗对照(P<0．05)。LOCM在本实验剂量不能诱导肾小管上皮细胞凋亡。HOCM在诱导HKC细胞凋亡时伴随有Bax、Bcl-2表达的上调,caspase-3活性升高。结论 HOCM和LOCM对肾小管上皮细胞均有毒性作用,LOCM的毒性作用明显小于HOCM。HOCM 可诱导人肾小管上皮细胞凋亡且与高渗及碘离子浓度有关。Bax／Bcl-2、caspase-3可能参与了造影剂诱导人肾小管上皮细胞凋亡的调控。
Objective To evaluate the cytotoxicity of high-osmolar contrast media （HOCM, diatrizoate） and low-osmolar contrast media （LOCM, iohexol） on human renal tubular epithelial cells （HKC）,and to determine the regulating roles of Bax/Bcl-2 and caspase-3 on apoptosis induced by contrast media （CM） in HKC. Methods HKC cells line was used. The subjects were divided into the following seven groups ： HOCM group （ 111 mg iodine/ml）, HOCM group （ 74 mg iodine/ml）, LOCM group（111 mg iodine/ml）, LOCM group（iohexol iodine/ml）, mannitol high-osmolar control group, mannitol low-osmolar control group and culture media control group. The cytotoxicity of HOCM and LOCM was evaluated by MTT and LDH assay. Apoptosis was assessed by Hochest33258 fluorescence stained cytospins, TUNEL staining, DNA agarose gel electrophoresis, electron microscope and flow cytometric DNA analysis. The protein expression of Bax/Bcl-2 was examined by Western blot, and caspase-3 activity was determined by fluorometric method. Results Compared to the control group, LDH levels increased significantly（P 〈 0.05） and cell viability decreased in HOCM or LOCM group （P 〈 0.05） with an osmotic pressure and iodinated ion in a time-deoendent manner.In HOCM groups, diatrizoate induced cultured HKC apoptosis. In LOCM groups, iohexol did not induced apoptosis. With mannitol under the same osmotic pressure, apoptosis increased in HKC incubated with diatrizoate（P〈0.05）. Bax, Bcl-2 production, and caspase-3 activity were upregulated in cultured HKC treated with 74 or 111 mg iodine/ml meglumine diatrizoate. Conclusions Both HOCM and LOCM have toxic effect on HKC cells. HOCM is more cytotoxic than LOCM. HOCM can induce cultured HKC cells apoptosis, which is related to the hypertonicity and iodine concentration of contrast media. Bax/Bcl-2 and caspase-3 may play an important role in mediating apoptosis in contrast media- induced cultured HKC.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2006年第2期88-93,共6页
Chinese Journal of Nephrology
基金
湖南省科委社会发展基金(No.1013-98)
