不同浓度硫化氢对肠上皮细胞炎症、线粒体功能和氧化应激的影响

Effects of different concentrations of hydrogen sulfide on inflammation, mitochondrial function and oxidative stress of intestinal epithelial cells

[目的]本试验使用硫氢化钠(NaHS,硫化氢供体)处理Caco-2细胞,研究硫化氢(H2S)对肠上皮细胞炎症、氧化应激和线粒体硫化物代谢功能的影响,探究H 2S影响肠道健康的可能机制。[方法]试验分为4个组,分别为对照组、低(0.1 mmol·L^-1)、中(1 mmol·L^-1)、高(2 mmol·L^-1)浓度的NaHS组,处理24 h后测定各组细胞活力、炎症细胞因子基因表达、硫化物代谢酶基因表达和氧化应激指标。[结果]与对照组相比,NaHS能够增强肠上皮细胞活力,2 mmol·L^-1 NaHS显著上调肿瘤坏死因子α(TNF-α)的基因表达水平(P<0.05),0.1 mmol·L^-1 NaHS上调线粒体硫化物代谢酶硫醌氧化还原酶(SQR)、硫代硫酸盐硫转移酶(TST)、硫双加氧酶(ETHE1)和亚硫酸氧化酶(SUOX)的基因表达,而2mmol·L-1 NaHS下调SQR的基因表达,显著提高丙二醛(MDA)、谷胱甘肽(GSH)含量和谷胱甘肽过氧化物酶(GPx)活性(P<0.05),显著降低超氧化物歧化酶(SOD)活性(P<0.05),0.1mmol·L^-1 NaHS显著提高MDA含量(P<0.05),1mmol·L^-1 NaHS则显著降低SOD活性(P<0.05)。[结论]低浓度H 2S能增强细胞活力,提高线粒体硫化物代谢能力和氧化应激水平。高浓度H 2S能促进炎症反应,提高氧化应激水平,影响线粒体硫化物代谢能力,可能对肠道健康产生不利影响。

[Objectives]The paper aimed to investigate the potential mechanism of hydrogen sulfide(H 2S)modulation on gut health.Caco-2 cells were treated with sodium hydrosulfide(NaHS,H 2S donor)and the effects of H 2S on inflammation,oxidative stress and mitochondrial sulfide metabolism in intestinal epithelial cells were studied.[Methods]The experiments included four groups:control group,low NaHS(0.1 mmol·L^-1)group,medium NaHS(1 mmol·L^-1)group,and high NaHS(2 mmol·L^-1)group.After 24 h treatment,cell viability,the gene expression of inflammatory cytokine and sulfide metabolism enzymes,and oxidative stress were measured.[Results]The results showed that NaHS enhanced cell viability.2 mmol·L^-1 NaHS significantly up-regulated the mRNA expression of tumor necrosis factor alpha(TNF-α)(P<0.05).0.1 mmol·L-1 NaHS up-regulated the gene expressions of the mitochondrial sulfide metabolism enzymes sulfide quinone oxidoreductase(SQR),thiosulfate sulfur transferase(TST),sulfur dioxygenase(ETHE1)and sulfite oxidase(SUOX).However,2 mmol·L-1 NaHS down-regulated gene expression of SQR.2 mmol·L-1 NaHS significantly increased malondialdehyde(MDA)content,glutathione(GSH)content,glutathione peroxidase(GPx)activity(P<0.05)and significantly reduced superoxide dismutase(SOD)activity(P<0.05).0.1 mmol·L^-1 NaHS significantly increased MDA content(P<0.05).1 mmol·L^-1 NaHS significantly reduced SOD activity(P<0.05).[Conclusions]Low concentration of H 2S enhances cell viability,mitochondrial sulfide metabolism and oxidative stress.High concentration of H 2S promotes inflammatory response and oxidative stress,and affects the mitochondrial sulfide metabolism ability,which may be detrimental to gut health.