55 results about "Intracellular glutathione" patented technology

The invention discloses a fluorescence probe for detecting glutathione and a preparation method and a use method of the fluorescence probe. A classical ICT system is constructed by utilizing 1,8-naphthalimides, a benzene sulfoxide part is introduced at 4-position, and the ICT effect of probe molecules is adjusted and controlled by utilizing a classical ICT system constructed by 1,8-naphthalimide and introducing benzene sulfoxide part at the fourth position. Under the condition that no glutathione exists, the probe molecules do not emit fluorescence light because of strong electron-withdrawing effect of 4-position benzene sulfoxide; under the condition that glutathione exists, the benzene sulfoxide part can be replaced by the glutathione, and thus intramolecular electron transfers from a 4-position sulphur atom (from the glutathione) to the 1,8-naphthalimides, and the probe molecules emit hyperfluorescence because of the ICT effect. According to the invention, the detection of the intracellular glutathione can be realized, and the fluorescence probe has the advantages of convenient operation, low cost, sensitive response, easy promotion and application and the like.

The invention discloses application of a withanolide compound in preparation of a drug or a healthcare product for treating or preventing tumors and neurodegenerative diseases. The withanolide compound has structural formulas as shown in formula I and optical isomers thereof. The research indicates that the withanolide compounds can be used for activating an Nrf2 signal channel, up regulating expression of NrF2 and downstream antioxidant and II-phase detoxifying enzymes thereof, strengthening the level of glutathione in cells, increasing reducing capacity in cells, promoting elimination of active carbon, restraining lipid peroxidation and lowering cytotoxicity caused by cancerogen arsenic and nerve cell toxicity induced by active oxygen H2O2 to show good chemical prevention effect. And therefore, the withanolide compound and nightshade extract including the withanolide compound have the potential of becoming a chemical prophylactic drug and the healthcare product, and can be used for preparing the corresponding oral preparation and injection by being used as the main ingredient, and also can be used for preventing and treating tumors and neurodegenerative diseases.

Food containing γ-glutamylcysteine or cysteine is produced by culturing Candida utilis containing 1% by weight or more of γ-glutamylcysteine per dry cells in logarithmic growth phase when cultured in a minimal medium, for example, Candida utilis in which a gene encoding glutathione synthetase is modified so that intracellular glutathione synthetase activity is reduced, under a suitable condition and mixing the obtained culture or a fraction thereof or the culture or a fraction thereof subjected to heat treatment with a raw material of food or drink to process food or drink.

The invention discloses a photodynamic induced CO releasing method, a CO controlled delivery system and a building method thereof. The CO releasing method is characterized by carrying out controlled release of CO under irradiation of near-infrared light by mixing a photosensitizer responding to the near-infrared light and applied to photodynamic therapy with carbon monoxide releasing molecules CORM-401. The CO controlled delivery system built on the basis of the photodynamic induced CO releasing method comprises a photosensitizer responding to near-infrared light and applied to photodynamic therapy, carbon monoxide releasing molecules CORM-401 and a carrier loaded or integrated with the photosensitizer and the carbon monoxide releasing molecules CORM-401. According to the CO controlled delivery system, the photochemical effect generated by the photosensitizer under induction of the near-infrared light is capable of accelerating the CO controlled delivery system to enter cells for decomposing the nanogel under the action of glutathione in the cells and releasing CORM-401; the near-infrared light activates and simultaneously generates singlet oxygen and CO; the singlet oxygen and COare respectively applied to photodynamic therapy and CO gas therapy; the combined therapy is achieved; the anti-tumor effect is obviously improved.

Dithiolane compounds having the structural formula (I):are useful for reinforcing and / or preserving the natural antioxidant protection of the skin against oxidative stress caused, especially, by UV radiation, e.g., by increasing the level of intracellular glutathione.

There is provided an oral composition capable of strimulating intracellular biosynthesis of glutathione in order to maintain in-vivo (i.e., in tissues and cells) glutathione level at a high level, by adding an active ingredient other than an amino acid precursor from a viewpoint of increasing intracellular glutathione synthesis, as a method which is an alternative or can be combined with a method of supplying an amino acid precursor to be used in glutathione biosynthesis. The oral composition is used for maintaining or increasing intracellular glutathione level and contains at least one rubrofusarin glycoside in an amount that is effective for increasing the intracellular glutathione.

According to the present invention, the following process for producing glutathione or gamma-glutamylcysteine is provided. A process for producing glutathione or gamma-glutamylcysteine by culturing in a medium a microorganism with a higher activity of one of the following proteins having an activity to transport intracellular glutathione to the outside of cells, and a higher activity of a protein involved in glutathione or -glutamylcysteine biosynthesis, compared with that of the parent strain, forming and accumulating glutathione or -glutamylcysteine in the medium, and recovering the glutathione or -glutamylcysteine from the culture. [1] A protein having the amino acid sequence shown by any of SEQ ID NOs: 1 to 26 [2] A protein consisting of an amino acid sequence wherein one or more amino acids have been deleted, substituted or added in the amino acid sequence shown by any of SEQ ID NOs: 1 to 26, and having glutathione transporting activity [3] A protein having 80% or more homology to the amino acid sequence shown by any of SEQ ID NOs: 1 to 26, and having glutathione transporting activity.

The present invention generally relates to the field of neurological, physiological and psychotic dysfunctions associated with a mental disorder such as schizophrenia, or a predisposition therefor. The invention further relates to genes and proteins, which, when varied in their normal expression, their nucleic acid sequence or in their activity, are associated with the mental disorder. Accordingly, the present invention relates to methods for diagnosis and to methods for prevention and / or treatment of a mental disorder. The present invention additionally relates to compositions for use in diagnosis and to kits for diagnosis of a mental disorder. The invention also relates to the use of a protein or polynucleotide for the manufacture of a medicament for use in the treatment and / or prevention and to a pharmaceutical composition for use in prevention and / or treatment of a mental disorder. Further, the invention relates to methods for screening for a modulator of a mental disorder. More particularly, the present invention relates to methods, compositions and kits, a microarray and reagents for determining the presence of at least one polymorphism and / or at least one combination of polymorphisms of at least one copy of a gene involved in regulating the intracellular glutathione (GSH) level and / or GSH-oxidative stress-related gene expression in a human being. Further, the invention relates to a pharmaceutical composition for use in the treatment and / or prevention of a mental disorder, to the use of an active ingredient such as a protein or polynucleotide for the manufacture of a medicament for use in the treatment and / or prevention of a mental disorder in patients with specific polymorphisms in genes involved in regulating the intracellular GSH level and / or GSH-oxidative stress-related gene expression. The invention also relates to methods of preventing and / or treating a mental disorder comprising administering a medicament to patients having said polymorphisms and to methods of screening for a modulator of a mental disorder.

The present invention provides neuroprotective polyphenol compounds, which can be synthetic analogs of fisetin, baicalein or chlorogenic acid, that maintain neuroprotective, anti-inflammatory, glutathione promoting, and / or antioxidant properties. The neuroprotective polyphenol compounds are useful for promoting, enhancing and / or increasing neuron protection, growth and / or regeneration. The polyphenol compounds further find use for increasing and or maintaining intracellular glutathione (GSH) levels. The polyphenol compounds are also useful for treating, preventing, mitigating and / or delaying neurodegenerative conditions, including diabetes, Parkinson's disease, Huntington's disease, Alzheimer's disease, non-Alzherimer's dementias, multiple sclerosis, traumatic brain injury, spinal cord injury or ALS.

The invention discloses a recombination strain secreting glutathionem, wherein the recombination strain has an inactivated gene of a transportation protein transporting the glutathionem from outside the cell to inside the cell, and expresses a gene of a transportation protein transporting exogenous glutathione from inside the cell to outside the cell. The recombination strain provided by the invention changes transportation pathways of the glutathione, and enables the synthesized glutathione to be secreted to outside of the cells, thereby greatly improving glutathione output, with a concentration of the extracellular glutathione being two times of that of the intracellular glutathione. The recombination strain provided by the invention has a total GSH production of more than 10g / L, and the GSH transported to outside of the cells has a production of more than 7g / L.

The invention discloses a microfluidic chip based laser induced fluorescence system for detecting the content of glutathione (GSH) in each cell and a method for rapidly and continuously detecting the content of GSH in each cell by adopting the microfluidic chip based laser induced fluorescence system. After derivatization from 2,3-naphthalenedicarboxaldehyde (NDA) is carried out in a cell, the derivative generated from GSH and NDA in the cell can emit fluorescent light after being irradiated with laser. The sample injection, separation and detection are carried out in a microfluidic chip. Four power supply output ends of an electric driver are respectively connected a buffer tank B, a sample tank S, a solution waste tank SW and a buffer waste tank BW of the microfluidic chip. The microfluidic chip is arranged on the detection platform of the microfluidic chip based laser induced fluorescence system. The fluorescent light emitted by the cell is acquired by a data acquisition device, and then the fluorescent light is displayed by the software of a computer. The method disclosed by the invention can be used for detecting the content of GSH in each cell without the necessity of breaking the cell. The microfluidic chip based laser induced fluorescence system has high sensitivity and high resolution. By adopting the method and the microfluidic chip based laser induced fluorescence system, the operation difficulty can be decreased, and the operation time can be shortened.

The invention provides a fluorescence imaging method for intracellular glutathione. In the method, a fluorescence sensor based on a controlled release technology is applied to fluorescence imaging of the intracellular glutathione. The adopted fluorescence sensor comprises a gold nanometer cage, a fluorescent dye and a glutathione recognition probe, and has the advantages of simple structure, skillful design, stable performance, high cell membrane permeability, high intracellular release controllability, high glutathione selectivity, high intracellular dispersity and the like. By adopting the fluorescence sensor, high-sensitivity and high-definition fluorescence imaging of the intracellular glutathione can be realized conveniently and rapidly, and the advantages of short response time, easiness in direct observation, convenience in real-time monitoring and the like are achieved. Based on the controlled release technology, a novel technology and a novel method are provided for targeted therapy, drug delivery, cell imaging and real-time monitoring of tumor cells.

The invention discloses a fullerene-based fluorescent probe with rapid and highly-efficient response to glutathione, and a preparation method and an application thereof. The fluorescent probe is a complex based on a fullerene quantum dot and a manganese dioxide nanosheet; the fluorescent probe with response to the glutathione is formed through the actions of electrostatic adsorption and coordination, and is expressed as FQD-MnO2; and the fullerene quantum dot has a structural formula of C60(OH)x(NH2)yOz. The fluorescent probe provided by the invention is simple in synthesis, has stable fluorescence, is insusceptible to quenching, can rapidly respond to the glutathione (GSH), and can be used for quantitative detection of the content of GSH in a solution and a cell. Under the condition of the presence of the GSH, the manganese dioxide nanosheet is rapidly reduced to manganese ions, and fluorescence of FQD is recovered at the same time. Through construction of the linear relationship between the concentration of the GSH and the fluorescence recovery intensity of the FQD, the concentration of the glutathione in the solution and the cell can be quantitatively detected. The fullerene-based fluorescent probe provided by the invention can efficiently and rapidly detect the content of the GSH and has high sensitivity and wide measurement concentration range.

The invention discloses novel application of a cinnamon extract, which relates to application of cinnamon (including cinnamon, chartophyllum, cinnamomum mollifolium, howood, cinnamomum burmannii and sassafras) extract to preparation of medicines or healthcare products for treating or preventing tumour. The invention also provides a preparation method of the cinnamon extract. Researches show that the cinnamon extract is capable of activating an Nrf2 signal path, promoting anti-oxidation and expression of II phase detoxifying enzymes, increasing the intracellular glutathione level, enhancing the intracellular reduction capability and inhibiting the cytotoxicity caused by cancerogen arsenic and reactive oxygen H2O2. Consequently, the cinnamon extract can be prepared into corresponding oral preparations and injections and used for preventing the tumour.

The invention discloses a method for preparing nanocluster gel of a hydroxyl-like phosphorite component. The method comprises the following steps: mixing a chloroauric acid solution and a adenine nucleotide series, reacting for 1 to 60 minutes, adding a citric acid buffering solution, standing for 0.5 to 24 hours in a light-shielding manner, adding absolute ethyl alcohol, uniformly mixing, and centrifuging to obtain precipitates, preparing the obtained precipitates into an adenine nucleotide shell nanocluster with the concentration of 0.1 to 10000 microgram / mL, adding metal ions to prepare sediments, dispersing the sediments in deionized water, thus obtaining the nanocluster gel. The fluorescent gold nanocluster adopting the adenine nucleotide series as ligand reacts with the metal ions toobtain aggregation-induced fluorescent nanocluster gel, the aggregation of a metal-phosphate structure achieves the fluorescent reinforced effect, the combination force of gold and adenine in the nanocluster gel is slightly weak, can be dissociated under the existence of molecules such as glutathione in the cell, the degradability is excellent, and the preparation method is simple in operation, high in yield, and low in cost.

The invention provides a polymer capable of releasing SO2, a preparation method and an application and nano-micell. The polymer has a structure shown in a formula I, the polymer material has amphipathy, and can be existed under cysteine or glutathione environment, a dinitrobenzene sulfonamide group in a formula I responses to a mercapto group, and is subjected to a rearrangement reaction to release SO2, and the material is subjected to self assembly in an aqueous solution to form the nano-micell. The polymer has good water dispersion performance, and can be endocytosed by cells, is capable ofreleasing SO2 under effect of glutathione in the cells, increases the active oxygen level in the cancer cells, and presents obvious killing effect for cancer cells. The polymer nanoparticles can reduce the cancer cell survival rate to 11+ / -1%, and the hydrodynamic radius of the nanoparticles is 29+ / -0.1 nm.

It is intended to provide a drug, a food or a feed which has an effect of enhancing the activity of a second-phase detoxifying enzyme and an effect of increasing intracellular glutathione content.

It is intended to provide a drug, a food or a feed which has an effect of enhancing the activity of a second-phase detoxifying enzyme and an effect of increasing intracellular glutathione content.

The invention discloses a tumor-specific cleavable PEG nanoparticles and a preparation method and application thereof. The invention modifies the surface of mesoporous silicon nanoparticles with cationic polymer layer containing amino group and disulfide bond, adds PEG containing benzaldehyde group at the end group, introduces PEG shell layer on the surface of the nanoparticles, and obtains tumor-specific cleavable PEG nanoparticles, wherein the surface of the mesoporous silicon nanoparticles is modified with cationic polymer layer containing amino group and disulfide bond, and then adds PEG containing benzaldehyde group at the end group. The carrier has the characteristics of PEG molecule shedding off in tumor microenvironment, responding to the redox of intracellular glutathione, strongdrug loading capacity, long blood circulation time, targeted drug delivery and other advantages, and is suitable for the preparation of anti-tumor drug carrier.

The invention discloses a high-yield glutathione pichia pastoris strain G3-SF and an application thereof. Pichia pastoris GS115 is taken as a host, Scgsh1 and Scgsh2 genes from saccharomyces cerevisiae are subjected to heterologous expression, excessive production of GSH is obtained, and an engineering bacterium is named as G3. On the basis, heterologous expression is derived from a saccharomyces cerevisiae adenosine kinase Scadk1 gene and a streptococcus sanguis derived from a codon optimized StgshF gene, and aims to enhance energy supply in an engineering bacterium fermentation process and reconstruct a synthesis path of GSH. The maximum yield of glutathione at a shake flask level can reach (527.14+ / -17.92) mg / L, and the intracellular glutathione yield is (19.96+ / -0.05) mg / L / OD. The constructed engineering bacterium G3-SA is fermented in a 5L fermentation tank, the highest yield of glutathione in 56h is 5950mg / L, the intracellular glutathione yield is 25.54 mg / L / OD, and a new thought is provided for industrial production of GSH.

The invention discloses an in-situ synthesis method of DNA silver nano-clusters Cells are subjected to DNA transfection by raising the level of glutathione (GSH) in cells, and therefore the cells are induced to synthesize the DNA silver nano-clusters in situ. The DNA silver nano-clusters synthesized in situ by the method have high biocompatibility, and the retention time of probes in the organisms is prolonged, so that the DNA silver nano-clusters can used for long-term monitoring over the activity of telomerase during the proliferation and differentiation processes of cells. In addition, by replacing sliver nano-clusers modified by oligonucleotide with different sequences as multifunctional fluorescent probes, the method can be used for detecting other functional molecules such as enzymes, glucose, protein, DNA and the like in a high-sensitivity mode and widely applied to the fields of bio-marking, disease monitoring and the like.

The invention discloses a photodynamic induced CO releasing method, a CO controlled delivery system and a building method thereof. The CO releasing method is characterized by carrying out controlled release of CO under irradiation of near-infrared light by mixing a photosensitizer responding to the near-infrared light and applied to photodynamic therapy with carbon monoxide releasing molecules CORM-401. The CO controlled delivery system built on the basis of the photodynamic induced CO releasing method comprises a photosensitizer responding to near-infrared light and applied to photodynamic therapy, carbon monoxide releasing molecules CORM-401 and a carrier loaded or integrated with the photosensitizer and the carbon monoxide releasing molecules CORM-401. According to the CO controlled delivery system, the photochemical effect generated by the photosensitizer under induction of the near-infrared light is capable of accelerating the CO controlled delivery system to enter cells for decomposing the nanogel under the action of glutathione in the cells and releasing CORM-401; the near-infrared light activates and simultaneously generates singlet oxygen and CO; the singlet oxygen and COare respectively applied to photodynamic therapy and CO gas therapy; the combined therapy is achieved; the anti-tumor effect is obviously improved.

The invention relates to the application of a morinin N or a salt thereof in the preparation of products for preventing and treating diseases related to ferroptosis. The present invention finds for the first time that Morinin N can effectively antagonize the reduction of glutathione level caused by ferroptosis or glutamic acid, effectively increase the content level of glutathione in cells, and can well improve and prevent ferroptosis or glutathione. Quercetin is significantly more active than the neuroprotective modulator quercetin in the related diseases induced by aminoacid, especially neurological injury diseases.

The invention relates to a method for improving the production of glutathione produced by fermentation of Candida utilis, which relates to a method for preparing glutathione by fermentation. The present invention adopts Candida utilis as the fermentation strain, after slant culture and seed culture, it is inoculated in the fermentation medium for shaking flask culture or fermentation tank culture, and L-cysteine ​​is added in the fermentation medium to increase the fermentation rate. The supply of L-cysteine ​​in the liquid can improve the synthesis speed and yield of the product glutathione. The advantage of the invention is that under the condition of other culture conditions being the same, the production and intracellular content of glutathione can be nearly doubled after adding L-cysteine ​​to the fermentation broth. This will greatly benefit the industrialized production of glutathione prepared by fermentation.

The invention discloses a method for determining the content of glutathione in bacterial cells. The method comprises the following steps: drafting a standard curve of different concentrations of glutathione to the fluorescence intensity; adopting a plate colony-counting method to carry out colony counting; setting a bacterial liquid experiment group for determination and a background fluorescence control group; processing the experiment group and the control group, and detecting by adopting a fluorescence intensity detection method; calculating according to the standard curve to obtain the glutathione content, and calculating according to flat counting to obtain the quantity of sample bacteria; and calculating a ratio of the glutathione content to the quantity of the corresponding bacteria to obtain the content of glutathione in a certain quantity of bacterial cells. An mBBr reagent is adopted in the invention, can go through the cell walls of Gram-positive bacteria, enters the cells, and combines with glutathione to generate a stable fluorescence derivative, and the bacteria are cracked by acetonitrile, so a problem of the wall breaking of the Gram-positive bacteria of the measurement method is solved.

The invention provides a method for releasing medicines by virtue of ester bond rupture. The method comprises the following steps: coupling small medicine molecules containing unsaturated carbanyl groups with tumor targeting polypeptide by virtue of ester bonds to obtain a polypeptide-small molecule compound, and then performing addition reaction on glutathione in cells and the polypeptide-small molecule compound to ensure that the ester bonds are broken, and the purpose of releasing the medicines can be achieved. According to the method for releasing the medicines by virtue of ester bond rupture, the small molecules containing the unsaturated carbanyl groups are coupled with fluorescence-labelled polypeptide molecules by virtue of the ester bonds to obtain a short peptide-small molecule compound. Inventors discover that the addition reaction can be performed between glutathione and the unsaturated carbanyl groups in the short peptide-small molecule compound to cause ester bond rupture so as to ensure that the medicines are released; and moreover, the ester bond rupture rate has a linear relationship with the concentration of glutathione, and when the polypeptide-small molecule compound enters the cells, the small molecules can be rapidly released, and the high sensitivity of the reaction on the concentration of glutathione is very suitable for selective release in the cells.

The invention discloses a construction method of a Grx-roGFP / Gpx3-roGFP gene overexpression 16HBE monoclonal cell line model. The construction method comprises the following steps: step 1, firstly, cloning target genes Grx-roGFP and Gpx-roGFP to a lentiviral vector pLV-puro; and 2, further packaging the lentivirus by using a lentiviral vector pLV-W, infecting 16HBE cells with the lentivirus, screening Grx-roGFP and Gpx-roGFP gene overexpression stable cell strains through puromycin, thereby constructing a cell line capable of dynamically observing the redox dynamic state in bronchial epithelial cell mitochondria in real time, and specifically detecting glutaredoxin (Glutaredoxin, Gpx-roGFP and Gpx-roGFP) of GSH. Grx is fused with glutathione peroxidase Gpx3 and roGFP for specifically detecting H2O2, and glutathione and H2O2 in cells can be dynamically detected in real time by observing the Grx-roGFP and the GPX3-roGFP, so that the change of the oxidation-reduction state in the cells can be speculated, the pathogenesis of active oxygen is disclosed, and a new foundation is laid for research on bacterial and virus infection and tobacco exposure. And lung lesions caused by dust and smoke, weather and air quality in some workplaces are provided with an in-situ model.