DeLeon Eric R, Gao Yan, Huang Evelyn, Olson Kenneth R
Indiana University School of Medicine-South Bend Center, South Bend, Indiana; and Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana.
Indiana University School of Medicine-South Bend Center, South Bend, Indiana; and.
Am J Physiol Regul Integr Comp Physiol. 2016 Jun 1;310(11):R1212-25. doi: 10.1152/ajpregu.00061.2016. Epub 2016 Apr 13.
The health benefits of garlic and other organosulfur-containing foods are well recognized and have been attributed to both prooxidant and antioxidant activities. The effects of garlic are surprisingly similar to those of hydrogen sulfide (H2S), which is also known to be released from garlic under certain conditions. However, recent evidence suggests that polysulfides, not H2S, may be the actual mediator of physiological signaling. In this study, we monitored formation of H2S and polysulfides from garlic oil in buffer and in human embryonic kidney (HEK) 293 cells with fluorescent dyes, 7-azido-4-methylcoumarin and SSP4, respectively and redox activity with two redox indicators redox-sensitive green fluorescent protein (roGFP) and DCF. Our results show that H2S release from garlic oil in buffer requires other low-molecular-weight thiols, such as cysteine (Cys) or glutathione (GSH), whereas polysulfides are readily detected in garlic oil alone. Administration of garlic oil to cells rapidly increases intracellular polysulfide but has minimal effects on H2S unless Cys or GSH are also present in the extracellular medium. We also observed that garlic oil and diallyltrisulfide (DATS) potently oxidized roGFP in buffer but did not affect DCF. This appears to be a direct polysulfide-mediated oxidation that does not require a reactive oxygen species intermediate. Conversely, when applied to cells, garlic oil became a significant intracellular reductant independent of extracellular Cys or GSH. This suggests that intracellular metabolism and further processing of the sulfur moieties are necessary to confer antioxidant properties to garlic oil in vivo.
大蒜和其他含有机硫食物对健康的益处已得到广泛认可,这归因于其促氧化和抗氧化活性。大蒜的作用与硫化氢(H₂S)惊人地相似,在某些条件下,大蒜也会释放出H₂S。然而,最近的证据表明,多硫化物而非H₂S可能是生理信号的实际介质。在本研究中,我们分别使用荧光染料7-叠氮基-4-甲基香豆素和SSP4监测缓冲液和人胚肾(HEK)293细胞中大蒜油产生H₂S和多硫化物的情况,并用两种氧化还原指示剂——氧化还原敏感的绿色荧光蛋白(roGFP)和二氯荧光素(DCF)监测氧化还原活性。我们的结果表明,缓冲液中大蒜油释放H₂S需要其他低分子量硫醇,如半胱氨酸(Cys)或谷胱甘肽(GSH),而仅在大蒜油中就能轻易检测到多硫化物。向细胞中添加大蒜油会迅速增加细胞内多硫化物,但对H₂S的影响极小,除非细胞外培养基中也存在Cys或GSH。我们还观察到,大蒜油和二烯丙基三硫化物(DATS)在缓冲液中能有效氧化roGFP,但不影响DCF。这似乎是一种直接的多硫化物介导的氧化,不需要活性氧中间体。相反,当应用于细胞时,大蒜油成为一种重要的细胞内还原剂,与细胞外的Cys或GSH无关。这表明,大蒜油在体内具有抗氧化特性需要细胞内对硫部分进行代谢和进一步加工。