Stress-related gene expression of HK-2 cells exposed to GF/FAF HSA, GF HSA, AngII or AngII+Cand. Homo sapiens

Proteinuria is pathogenic to proximal tubular cells (PTC) and linked with progression to renal failure. Angiotensin II (AngII) is also independently involved in the pathogenesis of progressive renal injury in varied kidney disease. The effects of human serum albumin (HSA) overload, AngII and candesartan, a specific inhibitor of AngII type 1 recptor, on the changes in gene protein expression stimulated by oxidative stress in PTC were assesed using cDNA microarrays. Keywords: stress response Overall design: Cells were growth arrested for 48 h in serum-free DMEM/Ham's F-12 medium with 5.5 mM glucose, 2 mM L-glutamine,100 U/ml penicillin, 100 ug/ml streptomycin, 20 mM Hepes. Media were refreshed and cells were incubated for 24h in conditioned media alone, or with media supplemented with 30mg/ml of different HSA preparations, 1 uM AngII or AngII in the presence of candesartan. After further 24 h under standard cell culture conditions (37C, 5% CO2) cells were subjected to RNA extraction.

蛋白尿(Proteinuria)对近端肾小管上皮细胞(proximal tubular cells, PTC)具有致病性，并与肾衰竭进展密切相关。血管紧张素II(Angiotensin II, AngII)亦可独立参与多种肾脏疾病中进行性肾损伤的发病过程。本研究采用cDNA微阵列(cDNA microarrays)技术，评估了人血清白蛋白(human serum albumin, HSA)负荷、AngII以及AngII 1型受体特异性抑制剂坎地沙坦(candesartan)对PTC中氧化应激诱导的基因蛋白表达变化的影响。关键词：应激反应(stress response) 总体实验设计：将细胞在含5.5 mM葡萄糖、2 mM L-谷氨酰胺、100 U/ml青霉素、100 μg/ml链霉素、20 mM Hepes的无血清DMEM/Ham's F-12培养基中进行48小时的生长阻滞。更换培养基后，将细胞分别置于仅含条件培养基、添加30mg/ml不同HSA制剂、1 μM AngII，或添加AngII与坎地沙坦的培养基中孵育24小时。随后在标准细胞培养条件（37℃、5% CO₂）下继续培养24小时，之后收集细胞进行RNA提取。