摘要
目的探讨血红素加氧酶1(heme oxygenase-1,HO-1)在高糖和晚期糖基化终末产物(AGE)诱导的人单核细胞氧化应激中的作用。方法根据是否加用15mmol/L葡萄糖(高糖)+100μg/ml AGE或HO-1抑制剂锌原卟啉(zinc protoporphyrin,ZnPP),将人单核细胞白血病细胞株THP-1细胞分为正常糖组(5mmoL/L)、高糖+AGE组、高糖+AGE+ZnPP组、正常糖+ZnPP组,孵育24h后收集细胞及培养液上清,检测各组细胞的活性氧簇(ROS)、培养液上清丙二醛和肿瘤坏死因子α(TNF—α)水平以及HO-1 mRNA和蛋白的表达。结果高糖+AGE组和正常糖+ZnPP组的ROS、丙二醛和TNF-α水平均显著高于正常糖组(均P〈0.05)。高糖+AGE+ZnPP组的ROS、TNF—α水平显著高于高糖+AGE组(均P〈0.05)。高糖+AGE+ZnPP组的HO-1 mRNA和蛋白表达水平显著低于高糖+AGE组(0.39±0.02vs0.89±0.09,0.38±0.00vs0.81±0.02,均P〈0.05)。结论ZnPP通过抑制HO-1表达加重高糖和AGE导致的单核细胞氧化应激,HO-1可能在糖尿病氧化应激中起重要作用。
Objective To investigate the effects of heine oxygenase-1 ( HO-1 ) on oxidative stress in THP-1 cells incubated with high glucose and advanced glycation end-products (AGE). Methods THP-1 cells were incubated with normal glucose ( 5 mmol/L), high glucose ( 15 mmol/L) +AGE ( 100 μg/ml), high glucose+AGE+ zinc protoporphyrin( ZnPP, inhibitor of HO-1 ), and ZnPP( normal glucose), respectively. The levels of reactive oxygen species(ROS) , malondialdehyde(MDA), and tumor necrosis factor-a(TNF-α) were determined in the four groups while expressions of HO-1 mRNA and protein were detected by RT-PCR and Western blotting respectively. Results The levels of ROS,MDA, and TNF-α in high glucose+AGE and ZnPP groups were higher than those in normal glucose group( all P〈0.05). The levels of ROS and TNF-α in high glucose+AGE+ ZnPP group were even higher compared with high glucose+AGE group( all P〈0.05 ) , along with decreased expressions of HO-1 mRNA and protein(0.39±0.02 vs 0. 89±0.09,0.38±0.00 vs 0. 81 ±0.02 ,both P〈0.05 ). Conclusions The oxidative injury and inflammation in THP-1 cells induced by high glucose and AGE are exaggerated by HO-1 inhibitor. HO-1 may play an important role in diabetic oxidative stress.
出处
《中华内分泌代谢杂志》
CAS
CSCD
北大核心
2009年第6期631-633,共3页
Chinese Journal of Endocrinology and Metabolism