Qi Hong-Na, Cui Jie, Liu Lei, Lu Fei-Fei, Song Cheng-Jie, Shi Yue, Yan Chang-Dong
Department of Physiology, Xuzhou Medical College, Xuzhou 221002, China.
Sheng Li Xue Bao. 2012 Aug 25;64(4):425-32.
The present study was aimed to investigate the effect of pretreatment with hydrogen sulfide (H2S) on human umbilical vein endothelial cells (HUVECs) senescence and the underlying mechanism. Cultured HUVECs at twelfth and fourth passages were taken as old and young groups, respectively. Sodium hydrosulfide (NaHS, donor of H2S) group was treated with NaHS from fourth to twelfth passage. The cell senescence was determined by senescence-associated β-galactosidase (SA β-gal) staining. DAPI fluorescent dye was used to detect cellular apoptosis. Western blot was used to analyze the expression levels of xanthine oxidase (XOD), manganese-superoxide dismutase (Mn-SOD) and the subunits p67(phox) of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in the HUVECs. Colorimetric method was used to detect SOD activity and cellular hydrogen peroxide (H2O2) level. The results showed that, compared with young group, the old group exhibited higher SA β-gal positive rate and cellular apoptosis, while NaHS pretreatment decreased SA β-gal positive rate and cellular apoptosis. Compared with the young group, the old group showed increased expression levels of XOD and p67(phox), as well as lower Mn-SOD expression level. With the pretreatment of NaHS, the up-regulations of XOD and p67(phox) levels and down-regulation of Mn-SOD level were inhibited. Compared with the young group, the old group showed lower SOD activity and higher H2O2 level, whereas NaHS pretreatment reversed the changes of SOD activity and H2O2 level. These results suggest that H2S delays senescence of HUVECs through lessening oxidative stress.
本研究旨在探讨硫化氢（H2S）预处理对人脐静脉内皮细胞（HUVECs）衰老的影响及其潜在机制。分别将第12代和第4代培养的HUVECs作为老年组和青年组。硫氢化钠（NaHS，H2S供体）组从第4代至第12代用NaHS处理。通过衰老相关β-半乳糖苷酶（SA β-gal）染色确定细胞衰老。用DAPI荧光染料检测细胞凋亡。采用蛋白质免疫印迹法分析HUVECs中黄嘌呤氧化酶（XOD）、锰超氧化物歧化酶（Mn-SOD）和烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶亚基p67(phox)的表达水平。采用比色法检测超氧化物歧化酶（SOD）活性和细胞过氧化氢（H2O2）水平。结果显示，与青年组相比，老年组SA β-gal阳性率和细胞凋亡率更高，而NaHS预处理降低了SA β-gal阳性率和细胞凋亡率。与青年组相比，老年组XOD和p67(phox)表达水平升高，而Mn-SOD表达水平降低。经NaHS预处理后，XOD和p67(phox)水平的上调以及Mn-SOD水平的下调受到抑制。与青年组相比，老年组SOD活性较低，H2O2水平较高，而NaHS预处理逆转了SOD活性和H2O2水平的变化。这些结果表明，H2S通过减轻氧化应激延缓HUVECs衰老。
