Effects of oxidative stress and its secondary protein impairment induced by high glucose content in human renal tubular epithelial cell / 解放军医学杂志

ABSTRACT

Objective To explore the effects of oxidative stress and its secondary protein impairment induced by high glucose content in human kidney proximal tubular epithelial cell line (HKC). Methods Cultured HKC were used to detect the reactive oxygen species (ROS) and protein carbonyl content. HKC were divided into three different groups control group (normal glucose), short-term treatment with high glucose group and long-term treatment with high glucose group. The glucose concentrations in control group and the high glucose treatment groups were 5.5mmol/L and 30mmol/L, respectively. Six time-points were set in short-term treatment with high glucose group namely 30min, 1h, 6h, 12h, 24h and 48h; glucose treatment was instituted for 2 months in long-term treatment of high glucose group. 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H2DCFDA) was used as ROS trapping agent, and flow cytometry and laser confocal microscopy were used to detect the concentration of ROS in HKC after treatment with high glucose for different periods of time. Flourescent probe JC-1 was used to test the change in mitochondrion membrane potential (MMP). The protein carbonyl content was determined by 2,4-dinitrobenzene hydralazine (DNPH) chromatometry. Results The ROS level in HKC reached the peak at 1 hour after treatment with high glucose, and it then decreased gradually (P0.05), but after 2 months, there was significant change in the content of DNPH (P