• The Korean Journal of Physiology and Pharmacology
• /
• v.7 no.3
• /
• pp.163-168
• /
• 2003

The reactive oxygen species (ROS) are considered to be an important mediator in pancreatic ${\beta}$ cell destruction, thereby triggering the development of insulin-dependent diabetes mellitus. In the present study, HIV-1 Tat-mediated transduction of Cu,Zn-superoxide dismutase (SOD) was investigated to evaluate its protective potential against streptozotocin (STZ)-induced cytotoxicity in insulin-producing MIN6N cells. Tat-SOD fusion protein was successfully delivered into MIN6N cells in a dose-dependent manner and the transduced fusion protein was enzymatically active for 48 h. The STZ induced-cell destruction, superoxide anion radical production, and DNA fragmentation of MIN6N cells were significantly decreased in the cells pretreated with Tat-SOD for 1 h. Furthermore, the transduction of Tat-SOD increased Bcl-2 and heat shock protein 70 (hsp70) expressions in cells exposed to STZ, which might be partly responsible for the effect of Tat-SOD. These results suggest that an increased of free radical scavenging activity by transduction of Tat-SOD enhanced the tolerance of the cell against oxidative stress in STZ-treated MIN6N cells. Therefore, this Tat-SOD transduction technique may provide a new strategy to protect the pancreatic ${\beta}$ cell destruction in ROS-mediated diabetes.

• Molecules and Cells
• /
• v.19 no.2
• /
• pp.191-197
• /
• 2005

The human immunodeficiency virus type 1 (HIV-1) Tat protein transduction domain (PTD) is responsible for highly efficient protein transduction across plasma membranes. In a previous study, we showed that Tat-Cu,Zn-superoxide dismutase (Tat-SOD) can be directly transduced into mammalian cells across the lipid membrane barrier. In this study, we fused the human SOD gene with a Tat PTD transduction vector at its N- and/or C-terminus. The fusion proteins (Tat-SOD, SOD-Tat, Tat-SOD-Tat) were purified from Escherichia coli and their ability to enter cells in vitro and in vivo compared by Western blotting and immunohistochemistry. The transduction efficiencies and biological activities of the SOD fusion protein with the Tat PTD at either terminus were equivalent and lower than the fusion protein with the Tat PTD at both termini. The availability of a more efficient SOD fusion protein provides a powerful vehicle for therapy in human diseases related to this anti-oxidant enzyme and to reactive oxygen species.

• BMB Reports
• /
• v.44 no.7
• /
• pp.462-467
• /
• 2011

Up-regulation of selected matrix metalloproteinases (MMPs) such as MMP-9 contributes to inflammatory processes during the development of various skin diseases, such as atopic dermatitis. In this study, we examined the effect of a cell-permeable superoxide dismutase (Tat-SOD) on TNF-${\alpha}$-induced MMP-9 expression in human keratinocyte cells (HaCaT). When Tat-SOD was added to the culture medium of HaCaT cells, it rapidly entered the cells in dose- and time-dependent manners. Tat-SOD decreased TNF-${\alpha}$-induced reactive oxygen species (ROS) generation. Tat-SOD also inhibited TNF-${\alpha}$-induced NF-${\kappa}B$ DNA binding activity. Treatment of HaCaT cells with Tat-SOD significantly inhibited TNF-${\alpha}$-induced mRNA and protein expression of MMP-9, as measured by RT-PCR and Western blot analysis. In addition, Tat-SOD suppressed TNF-${\alpha}$-induced gelatinolytic activity of MMP-9. Taken together, our results indicate that Tat-SOD can suppress TNF-${\alpha}$-induced MMP-9 expression via ROS-NF-${\kappa}B$-dependent mechanisms in keratinocytes, and therefore can be used as an immunomodulatory agent against inflammatory skin diseases related to oxidative stress.

• Proceedings of the SCSK Conference
• /
• 2003.09a
• /
• pp.630-644
• /
• 2003

A human Cu, Zn-superoxide dismutase (Cu, Zn-SOD) was fused with a Tat PTD of HIV-1 to produce a novel anti-aging ingredient, Tat-SOD for cosmeceuticals. Test of stability and evaluation of transduction efficacy and enzymatic activity suggest Tat-SOD is an effective active ingredient for anti-aging treatment.

• BMB Reports
• /
• v.44 no.12
• /
• pp.787-792
• /
• 2011

We investigated whether silk fibroin peptide derived from the silkworm, Bombyx mori, could inhibit inflammation and enhance the anti-inflammatory activity of Tat-superoxide dismutase (Tat-SOD), which was previously reported to effectively penetrate various cells and tissues and exert anti-oxidative activity in a mouse model of inflammation. Inflammation was induced by topical treatment of mouse ears with 12-O-tetradecanoylphorbol-13-acetate (TPA). Histological, Western blot, and reverse transcription-polymerase chain reaction data demonstrated that silk fibroin peptide or Tat-SOD alone could suppress elevated levels of cyclooxygenase-2, interleukin-6, interleukin-1beta, and tumor necrosis factor-alpha induced by TPA. Moreover, silk fibroin peptide significantly enhanced the anti-inflammatory activity of Tat-SOD, although it had no influence on in vitro and in vivo transduction of Tat-SOD. Silk fibroin peptide exhibited anti-inflammatory activity in a mice model of inflammation. Therefore, silk fibroin peptide alone or in combination with Tat-SOD might be used as a therapeutic agent for various inflammatory diseases.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.9
• /
• pp.1613-1619
• /
• 2008

It has been reported that Tat-SOD can be directly transduced into mammalian cells and skin and acts as a potential therapeutic protein in various diseases. To isolate the compound that can enhance the transduction efficiency of Tat-SOD, we screened a number of natural products. 3-O-[$\beta$-D-Glucopyranosyl(1$\rightarrow$4)-$\alpha$-L-arabinopyranosyll-hederagenin (OGAH) was identified as an active component of Fatsia japonica and is known as triterpenoid glycosides (hederagenin saponins). OGAH enhanced the transduction efficiencies of Tat-SOD into HeLa cells and mice skin. The enzymatic activities in the presence of OGAH were markedly increased in vitro and in vivo when compared with the controls. Although the mechanism is not fully understood, we suggest that OGAH, the active component of Fatsia japonica, might change the conformation of the membrane structure and it may be useful as an ingredient in anti-aging cosmetics or as a stimulator of therapeutic proteins that can be used in various disorders related to reactive oxygen species (ROS).

• BMB Reports
• /
• v.43 no.8
• /
• pp.561-566
• /
• 2010

Though protein transduction domains (PTDs) are well known for the delivery of exogenous therapeutic proteins into living cells, the overall low efficiency of transduction is a serious obstacle. We investigated the effect of bog blueberry anthocyanins (BBA) on protein transduction efficiency and found that BBA enhanced the transduction efficiencies of Tat-SOD fusion protein into HeLa cells and mice skin. The enzymatic activities in the cells and skin tissue in the presence of BBA were markedly increased compared to controls. Further, BBA did not demonstrate any cell toxicity at various concentrations. Although the mechanism is not fully understood, we suggest that BBA might alter the conformation of the membrane, which would indicate that BBA can be used as a protein transduction enhancer for the efficient delivery of therapeutic proteins for a variety of disorders.

• The Korean Journal of Food And Nutrition
• /
• v.20 no.4
• /
• pp.356-362
• /
• 2007

This study was carried to develop the new material of functional food which antioxidant of natural substances of sprouts. We compared antioxdiative activity and antioxidant substances exist in sprouts. Antioxidaive activities of sprouts were measured by total polyphenolic acid contents, electron donating activity(EDA), SOD-liked activity and hydroxyl radical scavenging ability. Total polyphenolic acid content was very higher in Rape sprouts extract(Rap) than other sprout extracts. Also Rape extract was showed the most excellent antioxdiative activity in SOD liked activity at 86.94%. The EDA was ordered Rape sprouts extract(Rap)>Tatsoi sprouts extract(Tat)>Broccoli sprouts extract(Bro)>Alfalfa sprout extract(Alf). Hydroxy radical scavenging ability was the most effective in Rape sprouts extract(Rap). Therefore we could be certain that Rape sprouts extract(Rap) was the most effective in antioxidative activity from sprouts.

• The Journal of Korean Medicine
• /
• v.26 no.3 s.63
• /
• pp.204-214
• /
• 2005

Objectives : The verity extract of the Gagam-Hyungbang-Gihwang-tang (GHG) was assessed to determine the mechanisms of its antioxidant activity. Methods : The fellowing effects were measured : GHG exhibited a concentration-treatment; scavenging ${\alpha},\;{\alpha}-diphenyl-\beta-picrylhydrazyl$ (DPPH) radical, linoleic acid oxidation in a thiocyanate assay system, and superoxide anion, hydroxyl radical-induced DNA nicking. We investigated mRNA levels such as superoxide-dismutase. Results : The GHG extract showed dose-dependent free radical scavenging activity, including DPPH radicals, hydroxyl radicals, and superoxide anion, using different systems. The GHG was also found to be effective in protecting plasmid DNA against the strand breakage induced by hydroxyl radicals in Fenton's reaction mixture. Furthermore, SOD-1 mRNA expression levels increased in tat hepatoma H4IIE cells Conclusions : We expect that GHG will to helpful to the development of antioxidant activity treatments.