Abstract：
Objective To investigate the protective effects of an angiotensin-converting enzyme inhibitor after inducing oxidative stress on keloid fibroblasts. Methods Primary keloid fibroblasts were isolated and cultured by enzyme digestion combined with the tissue adhesion method in vitro, and the third to fifth generations of cells were selected for the experiment. For 24 hours, keloid fibroblasts were treated with different concentrations of hydrogen peroxide. Different concentrations of angiotensin-converting enzyme inhibitor were added to the keloid fibroblast culture medium, and then the cells were treated with hydrogen peroxide for 24 hours. Results With the increase of hydrogen peroxide concentration, the growth of keloid fibroblasts was inhibited and the levels of malondialdehyde, superoxide dismutase, and reactive oxygen species increased gradually, accompanied by an increase in the expression of nicotinamide adenine dinucleotide phosphate oxidase and collagen I mRNA. The expression of nicotinamide adenine dinucleotide phosphate oxidase-mRNA in keloid fibroblasts and the formation of reactive oxygen species in keloid fibroblasts were induced by different concentrations of angiotensin II, and the most significant effect was at 10-5 mmol/mL. The effects of diphenyleneiodonium chloride (NOX inhibitor), N-acetylcysteine (reactive oxygen species inhibitor) and nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase) RNA treatment on angiotensin II-induced nicotinamide adenine dinucleotide phosphate oxidase and collagen I increased significantly. Hydrogen peroxide and angiotensin II alone or combined can induce NADPH oxidase and reactive oxygen species expression in keloid fibroblasts. When the angiotensin-converting enzyme inhibitor was added, the expression of NADPH oxidase and reactive oxygen species in keloid induced by hydrogen peroxide and angiotensin II could be inhibited. Conclusion Oxidative stress can lead to increased expression of reactive oxygen species, NADPH oxidase and collagen I in keloid fibroblasts, suggesting oxidative stress mediates the migration of human keloid fibroblasts and extracellular matrix synthesis.
摘要：
目的探讨血管紧张素转换酶抑制剂诱导氧化应激对瘢痕疙瘩成纤维细胞的保护作用。方法采用酶消化结合组织粘附法分离培养原代瘢痕疙瘩成纤维细胞,选择第三至第五代细胞进行实验。24小时,用不同浓度的过氧化氢处理瘢痕疙瘩成纤维细胞。在瘢痕疙瘩成纤维细胞培养基中加入不同浓度的血管紧张素转换酶抑制剂,然后用过氧化氢处理细胞24小时。结果随着过氧化氢浓度的增加,瘢痕疙瘩成纤维细胞的生长受到抑制，丙二醛水平受到抑制，超氧化物歧化酶,活性氧逐渐增多,伴随着烟酰胺腺嘌呤二核苷酸磷酸氧化酶和胶原蛋白ImRNA表达的增加。不同浓度血管紧张素Ⅱ诱导瘢痕疙瘩成纤维细胞中烟酰胺腺嘌呤二核苷酸磷酸氧化酶-mRNA的表达及瘢痕疙瘩成纤维细胞活性氧的形成,在10-5mmol/mL时效果最显著。氯化二苯基碘鎓（NOX抑制剂）的作用，N-乙酰半胱氨酸(活性氧抑制剂)和烟酰胺腺嘌呤二核苷酸磷酸氧化酶(NADPH氧化酶)RNA处理对血管紧张素Ⅱ诱导的烟酰胺腺嘌呤二核苷酸磷酸氧化酶和胶原Ⅰ显著增加。过氧化氢和血管紧张素II单独或联合可诱导瘢痕疙瘩成纤维细胞NADPH氧化酶和活性氧表达。当加入血管紧张素转换酶抑制剂时，过氧化氢和血管紧张素Ⅱ对瘢痕疙瘩中NADPH氧化酶和活性氧的表达有抑制作用。结论氧化应激可导致活性氧表达增加,瘢痕疙瘩成纤维细胞中的NADPH氧化酶和I型胶原,提示氧化应激介导人瘢痕疙瘩成纤维细胞的迁移和细胞外基质的合成。
