Raisa Nauli, Septelia I Wanandi, Mohamad Sadikin, Radiana D Antarianto, Sri Widia A Jusman
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引用次数: 0
摘要
本研究旨在分析血红素合成抑制对细胞红蛋白表达的影响及其与瘢痕疙瘩成纤维细胞活力和增殖的相关性。对瘢痕疙瘩成纤维细胞进行了原代培养。琥珀酰丙酮抑制瘢痕疙瘩成纤维细胞血红素合成。我们用比色法测定了氨基乙酰丙酸脱水酶(ALAD)的活性;qRT-PCR检测细胞红蛋白mRNA表达,ELISA和免疫细胞化学检测细胞红蛋白表达,MTT检测成纤维细胞活力;BrdU试验对成纤维细胞增殖的影响结果表明,经琥珀酰丙酮(SA、1、2.5和5 mM)处理的瘢痕疙瘩成纤维细胞的ALAD酶活性水平低于对照组。2.5 mM和5 mM SA培养的瘢痕疙瘩成纤维细胞中细胞红蛋白mRNA和蛋白表达水平显著低于对照和1 mM SA培养的瘢痕疙瘩成纤维细胞。随着SA浓度的升高,瘢痕疙瘩成纤维细胞的活力和增殖能力降低。综上所述,琥珀酰丙酮的使用浓度为1;2.5;5 mM引起ALAD酶活性降低,表明血红素合成受到抑制。抑制血红素合成可以影响细胞红蛋白的表达,这与瘢痕疙瘩成纤维细胞的活力和增殖有关。
Inhibition of ALA dehydratase activity in heme biosynthesis reduces cytoglobin expression which is related to the proliferation and viability of keloid fibroblasts.
The aim of this study was to analyze the effect of heme synthesis inhibition on cytoglobin expression and its correlation with keloid fibroblast viability and proliferation. The study was conducted on primary culture of keloid fibroblasts. Heme synthesis in keloid fibroblasts was inhibited using succinyl acetone. We measured amino levulinic acid dehydratase (ALAD) enzyme activity using a colorimetric method; cytoglobin mRNA expression using qRT-PCR, cytoglobin protein expression using ELISA and immunocytochemistry, fibroblast viability using the MTT test; and fibroblast proliferation using BrdU test. The results showed that the ALAD enzyme activity level was lower in the keloid fibroblasts treated with succinyl-acetone (SA, 1, 2.5, and 5 mM) than in the control. The cytoglobin mRNA and protein expressions level were significantly lower in the keloid fibroblasts cultured with 2.5 mM and 5 mM SA than in the control and 1 mM SA. The viability and proliferation of the keloid fibroblasts decreased when the SA concentration was increased. In conclusion, the use of succinyl acetone at a concentration of 1; 2.5; and 5 mM caused decrease ALAD enzyme activity which indicated the inhibition of the heme synthesis. Inhibition of heme synthesis can affect cytoglobin expression, which correlates with the viability and proliferation of keloid fibroblasts.
期刊介绍:
Journal of Clinical Biochemistry and Nutrition (JCBN) is
an international, interdisciplinary publication encompassing
chemical, biochemical, physiological, pathological, toxicological and medical approaches to research on lipid peroxidation, free radicals, oxidative stress and nutrition. The
Journal welcomes original contributions dealing with all
aspects of clinical biochemistry and clinical nutrition
including both in vitro and in vivo studies.