血红素加氧酶1在高糖和晚期糖基化终末产物诱导的THP-1细胞氧化应激中的作用

Role of heme oxygenase-1 in high glucose and advanced glycation end-products-induced oxidative stress in THP-1 cells

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.