摘要
目的 研究环孢素A (CsA)对鼠肾小管上皮细胞的毒性作用及病理学机制。方法 利用体外培养实验法 ,将鼠肾小管上皮 (NRK 5 2E)细胞 ,在体积分数 95 %O2 / 5 %CO2 、37℃的条件下培养 2 4h后 ,用不同浓度的CsA处理 4 8h。对细胞损伤的形态学、培养液中乳酸脱氢酶 (LDH)含量及细胞的脱落死亡进行观察和检测 ;细胞凋亡采用TUNEL染色法检测。结果 用CsA处理 4 8h后 ,NRK 5 2E细胞出现了空泡化、脱落以至大片死亡 ;培养基中LDH含量明显增高。同时 ,经CsA处理的部分NRK 5 2E细胞核出现了明显断片化 ,TUNEL染色阳性 ,发生了细胞的凋亡。CsA对NRK 5 2E细胞的这种毒性作用与使用剂量和时间呈正相关。结论 CsA对鼠肾小管上皮细胞的毒性作用较强。细胞凋亡可能是CsA致肾小管细胞毒性的重要病理学机制之一。
Objective To investigate and compare cytotoxic effects of cyclosporine A (CsA) treatment in kidney tubule epithelium cells (NRK-52E cells), and whether apoptosis is involved in the CsA-induced toxicity. Methods NRK-52E cells were cultured at 37?℃ in 95?% O 2/5?% CO 2, and were treated with different concentrations of CsA for 48?h.. The damage induced by CsA was characterized by an early cell detachment from culture (by direct counting of adherent cells), lactate dehydrogenase (LDH) release, and the changes in morphology were also observed by a phase-contrast microscope. Assay of apoptosis was done by TUNEL staining. Results CsA treatment induced significant damage in NRK-52E cells, which was demonstrated by obvious signs of cell injury with changes in cell shape, aggregates, and cell loss. A time- and dose-dependent increase in LDH release was also observed for CsA treatment . More than 70?% of the NRK-52E cells treated with 20?μg/ml CsA for 48?h were dead. The apoptotic DNA fragments were observed in a population of NRK-52E cells treated with CsA by TUNEL staining, which represented the capacity of CsA to induce apoptosis in NRK-52E cells. Conclusions CsA treatment induced a time- and dose-dependent damage in NRK-52E cells, in which the apoptotic DNA fragments were also observed by TUNEL staining. Thus, the results obtained from the present study suggested that apoptosis, as a possible mechanism, might be involved in the CsA-induced tubulopathy.
出处
《中华器官移植杂志》
CAS
CSCD
北大核心
2003年第6期345-347,共3页
Chinese Journal of Organ Transplantation
