Antiviral Decoction of Isatidis Radix (板藍根 bǎn lán gēn) Inhibited Influenza Virus Adsorption on MDCK Cells by Cytoprotective Activity

AbstractThe aim of this study is to elucidate how the Isatidis Radix (板藍根 bǎn lán gēn) tonic, as an aqueous mixture of hundreds of compositions, interrupts the infection of influenza viruses to their host cells. The efficacy of the tonic was evaluated and expressed as cell proliferation rate and plaque reduction rate in Madin-Darby Canine Kidney (MDCK) cells, against 3 strains of influenza A and B viruses. This boiling water (at 100°C) extract of Isatidis Radix (RIE) showed antiviral activity against influenza virus A and B. The concentration for 50% inhibition of influenza virus A replication (IC50) in MDCK cell was 12.6mg/mL with a therapeutic index >8. When cells were incubated with RIE prior to virus adsorption, the numbers of viable cell were at least doubled compared to the numbers of virus control, RIE incubation after virus adsorption and RIE incubation with virus prior to adsorption, in both influenza virus A and B. Moreover, much less virus particles were spotted by scanning electron microscope (SEM) in the RIE pre-treated cells than the cells without RIE treatment. These results indicate the antiviral activity of RIE is mainly attributed to its host cell protection effect but not actions on virus or post-virus-adsorption interruption. Cell, but not virus, is more likely to be the action target of RIE

Hypothesis Paper Effects of Intracellular Superoxide Removal at Acupoints with TAT-SOD on Obesity

TAT-SOD is a recombinant protein of superoxide dismutase fused with TAT peptide. By pure accident, we discovered that topical application of TAT-SOD to acupoints could result in acupuncture-like action. This study aimed to validate the accidental discovery by investigating the effect on simple obesity of the topical application of TAT-SOD to acupoints in comparison with acupuncture. 90 subjects were divided into 3 groups for 12-week treatments. Regular hospital acupuncture treatment was given to Acupuncture Group 3 times a week. TAT-SOD Group were instructed first to locate acupoints and apply 0.1 ml of 5000u SOD/ml TAT-SOD cream in an area of 1 cm 2 to each of the same set of acupoints, which they then conducted at home three times daily. Placebo Group applied the vehicle cream the same manner as TAT-SOD Group. Both TAT-SOD and acupuncture treatments decreased adiposity with overall clinical effective rates of 60.0% and 76.7%, respectively. The placebo group showed no improvement. The results validate that the enzymatic removal of the intracellular superoxide at acupoints could generate acupuncture-like effects, and indicate a possibility of the new method as a simple substitute to acupuncture and an insight of superoxide modulation along meridians for acupuncture mechanism. © 2011 Elsevier Inc. All rights reserved. Introduction Superoxide is one of the main reactive oxygen species (ROS) in the cell, which act as second messengers in the intracellular signaling pathways involved in activation of proinflammatory responses and mediate degradation of aggregan and collagen Although a variety of exogenous anti-oxidant enzymes are available to protect against oxidative stresses, the major problem in using these enzymes is that they can not be delivered into cells, thus resulting in their inability to detoxify intracellular RO

In Vivo Radioprotective Activity of Cell-Permeable Bifunctional Antioxidant Enzyme GST-TAT-SOD against Whole-Body Ionizing Irradiation in Mice

GST-TAT-SOD was the fusion of superoxide dismutase (SOD), cell-permeable peptide TAT, and glutathione-S-transferase (GST). It was proved to be a potential selective radioprotector in vitro in our previous work. This study evaluated the in vivo radioprotective activity of GST-TAT-SOD against whole-body irradiation. We demonstrated that intraperitoneal injection of 0.5 ml GST-TAT-SOD (2 kU/ml) 2 h before the 6 Gy whole-body irradiation in mice almost completely prevented the splenic damage. It could significantly enhance the splenic antioxidant activity which kept the number of splenic white pulp and consequently resisted the shrinkage of the spleen. Moreover, the thymus index, hepatic antioxidant activity, and white blood cell (WBC) count of peripheral blood in irradiated mice pretreated with GST-TAT-SOD also remarkably increased. Although the treated and untreated irradiated mice showed no significant difference in the growth rate of animal body weight at 7 days postirradiation, the highest growth rate of body weight was observed in the GST-TAT-SOD-pretreated group. Furthermore, GST-TAT-SOD pretreatment increased resistance against 8 Gy whole-body irradiation and enhanced 30 d survival. The overall effect of GST-TAT-SOD seemed to be a bit more powerful than that of amifostine. In conclusion, GST-TAT-SOD would be a safe and potentially promising radioprotector

Efficient internalization of TAT peptide in zwitterionic DOPC phospholipid membrane revealed by neutron diffraction

The aim of this study is to investigate the interactions between TAT peptides and a neutral DOPC bilayer by using neutron lamellar diffraction. The distribution of TAT peptides and the perturbation of water distribution across the DOPC bilayer were revealed. When compared to our previous study on an anionic DOPC/DOPS bilayer (X. Chen et al., Biochim Biophys Acta. 2013. 1828 (8), 1982\u20131988), a much deeper insertion of TAT peptides was found in the hydrophobic core of DOPC bilayer at a depth of 6.0 \uc5 from the center of the bilayer, a position close to the double bond of fatty acyl chain. We conclude that the electrostatic attractions between the positively charged TAT peptides and the negatively charged headgroups of phospholipid are not essential for the direct translocation. Furthermore, the interactions of TAT peptides with the DOPC bilayer were found to vary in a concentration-dependent manner. A limited number of peptides first associate with the phosphate moieties on the lipid headgroups by using the guanidinium ions pairing. Then the energetically favorable water defect structures are adopted to maintain the arginine residues hydrated by drawing water molecules and lipid headgroups into the bilayer core. Such bilayer deformations consequently lead to the deep intercalation of TAT peptides into the bilayer core. Once a threshold concentration of TAT peptide in the bilayer is reached, a significant rearrangement of bilayer will happen and steady-state water pores will form.Peer reviewed: YesNRC publication: Ye

Development of a Novel Restrictive Medium for Monascus Enrichment From Hongqu Based on the Synergistic Stress of Lactic Acid and Ethanol

Hongqu is a famous fermented food produced by Monascus and has been used as food coloring, wine starters and food additives for thousands of years in China. Excellent Monascus strain is an important prerequisite for producing high-quality Hongqu. However, the isolation of Monascus pure culture from Hongqu samples is time-consuming and laborious because it is easily interfered by other microorganisms (especially filamentous fungi). Therefore, the development of restrictive medium for Monascus enrichment from Hongqu is of great significance for the preparation and screening of excellent Monascus strains. Results of this study showed that Monascus has good tolerance to lactic acid and ethanol. Under the conditions of tolerance limits [7.5% lactic acid (v/v) and 12.0% ethanol (v/v)], Monascus could not grow but it still retained the vitality of spore germination, and the spore activity gradually decreased with the increasing concentrations of lactic acid and ethanol. More interestingly, the addition of lactic acid and ethanol significantly changed the microbial community structure in rice milk inoculated with Hongqu. After response surface optimization, Monascus could be successfully enriched without the interference of other microorganisms when 3.98% (v/v) lactic acid and 6.24% (v/v) ethanol were added to rice milk simultaneously. The optimal enrichment duration of Monascus by the restrictive medium based on the synergistic stress of lactic acid and ethanol is 8∼24 h. The synergistic stress of lactic acid and ethanol had no obvious effects on the accumulation of major metabolites in the progeny of Monascus, and was suitable for the enrichment of Monascus from different types of Hongqu. Finally, the possible mechanisms on the tolerance of Monascus to the synergistic stress of lactic acid and ethanol were preliminarily studied. Under the synergistic stress of lactic acid and ethanol, the cell membrane of Monascus defends against lactic acid and ethanol into cells to some extent, and the superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) activities of Monascus were higher than those of other fungi, which significantly reduced the degree of lipid peroxidation of cell membrane, while secreting more amylase to make reducing sugars to provide the cells with enough energy to resist environmental stress. This work has great application value for the construction of Monascus strain library and the better development of its germplasm resources