白蛋白对近端肾小管上皮细胞增殖与凋亡的影响论文.docVIP

　　白蛋白对近端肾小管上皮细胞增殖与凋亡的影响论文 孙良忠， 岳智慧， 陈述枚 【摘要】 【目的】 探讨白蛋白对近端肾小管上皮细胞（PTCs）增殖与凋亡的影响。【方法】 用不同剂量（0.1，0.5，1，5，10，30 mg/mL）去脂牛血清白蛋白(dBSA)超载体外培养的NRK52E.freelg/mL作为对照。细胞增殖活性用MTS比色法检测和显微镜观察。细胞凋亡检测：荧光染料DAPI染核，DNA梯带实验，原位末端标记（TUNEL）实验和苔盼蓝染色计数。【结果】 高剂量dBSA超载未完全融合NRK52E细胞144 h后细胞变得更密集。MTS比色结果发现，高剂量dBSA（5、10、30 mg/mL）超载可明显诱导NRK52E细胞增殖，增殖活性分别为（0.700 ± 0.045）A、（1.021 ± 0.029） A和（1.273 ± 0.173） A，较对照细胞（0.441 ± 0.020） A显著升高，P均 0.05。高剂量dBSA超载的NRK52E细胞，有较多出现核碎裂和核固缩，有较强DNA梯带和原位末端标记信号。苔盼蓝染色计数发现，高剂量dBSA（5、10、30 mg/mL）超载NRK52E细胞72 h的死亡细胞数（680 000 ± 28 618）/mL， （970 000 ± 52 915）/mL和（1 132 500 ± 88 954）/mL较对照的死亡细胞数［（312 500 ± 28 395）/mL显著升高， P均 0.05；而且死亡细胞数呈剂量依赖性。【结论】白蛋白超载NRK52E细胞既可诱导细胞增殖，又可诱导细胞凋亡。 【关键词】 白蛋白; 肾小管上皮细胞; 细胞增殖; 细胞凋亡 Abstract: 【Objective】 To study the effects of albumin overload on proliferation and apoptosis in proximal tubular epithelial cells (PTCs). 【Methods】 Different concentration of delipidated bovine serum albumin (dBSA) ranged from 0.1-30 mg/mL g/mL as control. Cell proliferation icroscope and measured by One Solution Cell Proliferation Assay Kit (MTS assay). Apoptosis ethods: DNA laddering assay, In situ terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling (TUNEL assay), nuclei changes munofluorecent microscope after stained idino -2-phenylindole) and total death cells icroscope. 【Results】Under light microscope, sub-confluent NRK52E cells became more confluent after dBSA overload 144 h. MTS assay revealed that higher dosages of dBSA (5, 10, and 30 mg/mL) overload induced significant proliferation (0.700 ± 0.045A, 1.021 ± 0.029 A, and 1.273 ± 0.173 A, respectively) 48 hours later in NRK52E, and g/mL) overload NRK52E cells, there ore condensed or broken nuclei, stronger DNA ladders and more positive signals in TUNEL assay than those in log/mL) overload NRK52E (680 000 ± 28 618/mL, 970 000 ± 52 915/mL, and 1 132 500 ± 88 954/mL, respectively) L), P 0.05; And death cells increased in a dose-dependent manner after dBSA overload. 【Conclusion】 Albumin o