白蛋白对近端肾小管上皮细胞增殖与凋亡的影响  

Effects of Albumin on Proliferation and Apoptosis in Proximal Renal Tubular Cells

作　　者：孙良忠[1] 岳智慧[1] 陈述枚[1] 

机构地区：[1]中山大学附属第一医院儿科,广东广州510080

出　　处：《中山大学学报（医学科学版）》2007年第6期649-653,共5页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省自然科学基金(06300772);广东省医学科研基金(B2004033)

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(0.1,0.5,1,5,10,30mg/mL)去脂牛血清白蛋白(dBSA)超载体外培养的NRK52E,dBSA0mg/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144h后细胞变得更密集。MTS比色结果发现,高剂量dBSA(5、10、30mg/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、30mg/mL)超载NRK52E细胞72h的死亡细胞数[(680000±28618)/mL,(970000±52915)/mL和(1132500±88954)/mL较对照的死亡细胞数[(312500±28395)/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。[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 were added to NRK52E cells cultured in vitro; dBSA 0 mg/mL as control. Cell proliferation was observed under microscope and measured by One Solution Cell Proliferation Assay Kit （MTS assay）. Apoptosis were detected by following methods： DNA laddering assay, In situ terminal deoxynucleotidyl transferase （TdT）-mediated dUTP nick-end labeling （TUNEL assay）, nuclei changes were observed under immunofluorecent microscope after stained with DAPI （4＇,6-diamidino -2-phenylindole） and total death cells were counted with trypan blue staining under light microscope. [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 were higher than that of control cells （0.441 ± 0.020 A）, P〈 0.05. In higher dosages of dBSA （5, 10, and 30 mg/mL） overload NRK52E cells, there were more condensed or broken nuclei, stronger DNA ladders and more positive signals in TUNEL assay than those in lower dosages of dBSA overload or control NRK52E cells. Trypan blue staining revealed that death cells in higher dosages of dBSA （5, 10, and 30 mg/mL） overload NRK52E （680 000 ± 28 618/mL, 970 000 ± 52 915/mL, and 1 132 500 ± 88 954/mL, respectively） were higher than that of control cells （312 500 ± 28 395/mL）, P〈 0.05; And death cells increased in a dose-dependent manner after dBSA overload. [Conclusion] Albumin overload could induce proliferation and apoptosis in PTCs.

关 键 词：白蛋白 肾小管上皮细胞 细胞增殖 细胞凋亡 

分 类 号：R692.6[医药卫生—泌尿科学]