Glutathione Disulfide Liposomes - a Research Tool for the Study of Glutathione Disulfide Associated Functions and Dysfunctions.
Glutathione Disulfide Liposomes - a Research Tool for the Study of Glutathione Disulfide Associated Functions and Dysfunctions.
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DOI：
10.1016/j.bbrep.2016.06.017
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发表时间：
2016-09
影响因子：
2.7
通讯作者：
Guan X
中科院分区：
文献类型：
--
作者：
Sadhu SS;Xie J;Zhang H;Perumal O;Guan X
谷胱甘肽二硫化物(GSSG)是谷胱甘肽(GSH)的氧化形式。GSH是一种以毫米波浓度存在于生物系统中的三肽，是体内主要的抗氧化剂。GSSG的增加反映了细胞内氧化应激的增加，并与疾病状态有关。这种增加还被证明导致蛋白质S-谷胱甘肽基化的增加，这可以影响蛋白质的结构和功能。蛋白质S-谷胱甘肽基化在细胞氧化应激过程中起调节作用。虽然GSSG已经商业化，但由于GSSG不是细胞膜通透性的原因，以及缺乏特异性增加细胞内GSSG的方法，GSSG在GSSG相关的各种正常/异常生理功能中的作用尚未完全阐明。我们已经开发出能够有效地将GSSG输送到细胞内的阳离子脂质体。可以方便地制备不同浓度的GSSG脂质体。在1 mg/mLGSSG脂质体作用4h后，GSSG细胞内GSSG含量增加27.1±6.9倍(n=3)，细胞内蛋白质S-谷胱甘肽基化水平显著增加，证实了GSSG的功能效应。台盼蓝实验表明，GSSG脂质体对细胞无毒性，细胞存活率大于95%。稳定性研究表明，在−80℃下，干态GSSG脂质体的稳定性至少为70d。为了将谷胱甘肽二硫化物(GSSG)输送到细胞内，研制了GSSG脂质体。GSSG脂质体可使细胞内GSSG增加27.1±6.9倍。通过蛋白质S-谷胱甘肽酰化反应证实了所传递的GSSG具有功能有效性。
Glutathione disulfide (GSSG) is the oxidized form of glutathione (GSH). GSH is a tripeptide present in the biological system in mM concentration and is the major antioxidant in the body. An increase in GSSG reflects an increase in intracellular oxidative stress and is associated with disease sates. The increase has also been demonstrated to lead to an increase in protein S-glutathionylation that can affect the structure and function of proteins. Protein S-glutathionylation serves as a regulatory mechanism during cellular oxidative stress. Though GSSG is commercially available, its roles in various GSSG-associated normal/abnormal physiological functions have not been fully delineated due to the reason that GSSG is not cell membrane permeable and a lack of method to specifically increase GSSG in cells. We have developed cationic liposomes that can effectively deliver GSSG into cells. Various concentrations of GSSG liposomes can be conveniently prepared. At 1 mg/mL, the GSSG liposomes effectively increased intracellular GSSG by 27.1±6.9 folds (n=3) in 4 h and led to a significant increase in protein S-glutathionylation confirming that the increased GSSG is functionally effective. The Trypan blue assay demonstrated that GSSG liposomes were not cytotoxic; the cell viability was greater than 95% after cells were treated with the GSSG liposomes for 4 h. A stability study showed that the dry form of the GSSG liposomes were stable for at least 70 days when stored at −80 °C. Our data demonstrate that the GSSG liposomes can be a valuable tool in studying GSSG-associated physiological/pathological functions. Glutathione disulfide (GSSG) liposomes were developed to deliver GSSG into cells. The GSSG liposomes effectively increased intracellular GSSG by 27.1±6.9 folds. The delivered GSSG is confirmed to be functionally effective through protein S-glutathionylation.