WO2022176435A1 - アルコール性肝障害抑制用の水素水 - Google Patents
アルコール性肝障害抑制用の水素水 Download PDFInfo
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- WO2022176435A1 WO2022176435A1 PCT/JP2022/000524 JP2022000524W WO2022176435A1 WO 2022176435 A1 WO2022176435 A1 WO 2022176435A1 JP 2022000524 W JP2022000524 W JP 2022000524W WO 2022176435 A1 WO2022176435 A1 WO 2022176435A1
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- water
- hydrogen
- ethanol
- hydrogen water
- electrolyzed
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 221
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 title claims abstract description 185
- 239000001257 hydrogen Substances 0.000 title claims abstract description 175
- 229910052739 hydrogen Inorganic materials 0.000 title claims abstract description 175
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- 206010067125 Liver injury Diseases 0.000 claims description 21
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/46—Treatment of water, waste water, or sewage by electrochemical methods
- C02F1/461—Treatment of water, waste water, or sewage by electrochemical methods by electrolysis
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F1/00—Treatment of water, waste water, or sewage
- C02F1/68—Treatment of water, waste water, or sewage by addition of specified substances, e.g. trace elements, for ameliorating potable water
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/02—Non-contaminated water, e.g. for industrial water supply
- C02F2103/026—Treating water for medical or cosmetic purposes
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2201/00—Apparatus for treatment of water, waste water or sewage
- C02F2201/46—Apparatus for electrochemical processes
- C02F2201/461—Electrolysis apparatus
- C02F2201/46105—Details relating to the electrolytic devices
- C02F2201/46115—Electrolytic cell with membranes or diaphragms
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E60/00—Enabling technologies; Technologies with a potential or indirect contribution to GHG emissions mitigation
- Y02E60/30—Hydrogen technology
- Y02E60/36—Hydrogen production from non-carbon containing sources, e.g. by water electrolysis
Definitions
- the present invention relates to hydrogen water for suppressing alcoholic liver injury.
- alcoholic liver injury has been reported to cause various disorders (for example, from mild disorders such as hangovers to severe disorders such as coma and stem cell necrosis), and from the viewpoint of suppressing these liver disorders, Alcoholic liver injury inhibitors containing various active ingredients are used.
- this alcoholic liver injury inhibitor examples include, for example, inhibitors containing lysine and citric acid as active ingredients (see, for example, Patent Document 1), preventive compositions containing proline or lysine (see, for example, Patent Document 2). ), and an inhibitor containing gelatin or soluble collagen as an active ingredient (see, for example, Patent Document 3).
- JP 2007-161642 A JP-A-6-116144 JP-A-6-247876
- the present invention has been made in view of the above points, and is capable of effectively suppressing alcoholic liver injury, and is highly safe, easy to prepare, and inexpensive. of hydrogen water.
- the hydrogen water for suppressing alcoholic liver injury of the present invention is hydrogen water for suppressing alcoholic liver injury that is mixed with ethanol, and has a dissolved hydrogen concentration of 550 to 5600 ppb, The concentration of ethanol in hydrogen water when mixed with ethanol is 1 to 4%.
- the present invention it is possible to provide hydrogen water for suppressing alcoholic liver injury, which is highly safe, easy to prepare and inexpensive, and can effectively suppress alcoholic liver injury.
- FIG. 1 is a configuration diagram schematically showing the configuration of an electrolyzed water generating device for generating electrolyzed hydrogen water according to an embodiment of the present invention
- the hydrogen water of the present invention is hydrogen water for suppressing alcoholic liver injury that is mixed with ethanol, has a dissolved hydrogen concentration of 550 to 5600 ppb, and has an ethanol concentration of 1 to 1 in the hydrogen water when mixed with ethanol. It is 4% hydrogen water.
- HepG2 human liver cancer-derived model cells
- the dissolved hydrogen in the hydrogen water of the present invention reduces the activity of ADH (alcohol dehydrogenase), which is an enzyme that produces aldehyde from ethanol, so that the production of aldehyde by decomposition of ethanol in the liver is suppressed. It is thought that alcoholic liver injury is suppressed because cell damage (mitochondrial damage and DNA damage) caused by aldehyde cytotoxicity is reduced.
- ADH alcohol dehydrogenase
- the dissolved hydrogen in the hydrogen water of the present invention improves the activity of ALDH (aldehyde dehydrogenase)
- ALDH aldehyde dehydrogenase
- the metabolism of aldehyde decomposition into acetic acid by oxidation of aldehyde
- cytotoxicity of aldehyde It is thought that alcoholic liver injury is suppressed because the cell damage (mitochondrial damage and DNA damage) caused by the drug is reduced.
- the concentration of ethanol in the hydrogen water is less than 1%, the concentration of ethanol is too low, and the effect of suppressing alcoholic liver injury described above may not be obtained sufficiently.
- the above-mentioned effect of suppressing alcoholic liver injury may decrease.
- the dissolved hydrogen concentration in the hydrogen water is preferably 1100 to 1300 ppb.
- electrolyzed hydrogen water of the present invention for example, electrolyzed hydrogen water produced by subjecting water to electrolysis treatment can be used.
- FIG. 1 is a configuration diagram schematically showing the configuration of an electrolyzed water generator for generating electrolyzed hydrogen water according to this embodiment.
- the electrolyzed water generator 1 includes a plurality of electrolytic cells 3 and 4, and FIG. 1 shows the electrolyzed water generator 1 including a pair of electrolytic cells 3 and 4. In addition, the electrolyzed water generator 1 may include three or more electrolytic cells 3 and 4 .
- the electrolytic baths 3 and 4 are connected in series, and the electrolytic bath 3 is provided upstream with respect to the electrolytic bath 4 .
- the electrolytic cell 3 includes an electrolytic chamber 30 for electrolyzing water; It has a diaphragm 33 that separates the first electrode chamber 30A on the 31 side and the second electrode chamber 30B on the second feeder 32 side.
- One of the first power feeder 31 and the second power feeder 32 is applied as an anode power feeder, and the other is applied as a cathode power feeder. Water is supplied to both the first electrode chamber 30A and the second electrode chamber 30B of the electrolysis chamber 30, and a DC voltage is applied to the first power feeder 31 and the second power feeder 32, so that water is supplied to the electrolysis chamber 30. electrolysis occurs.
- a solid polymer membrane made of a fluorine-based resin having a sulfonic acid group is used for the diaphragm 33 of the electrolytic chamber 30 on the upstream side.
- a plating layer made of platinum is formed on both surfaces of the diaphragm 33 .
- a net-like metal such as expanded metal made of titanium or the like and having a platinum plating layer formed on the surface thereof is applied for the first power feeder 31 and the second power feeder 32.
- Such mesh-like first power feeder 31 and second power feeder 32 sandwich the diaphragm 33 and allow water to spread over the surface of the diaphragm 33 , thereby promoting electrolysis in the electrolysis chamber 30 .
- the plated layer of the diaphragm 33 and the first power feeder 31 and the second power feeder 32 are in contact and electrically connected.
- the diaphragm 33 allows ions generated by the electrolysis to pass therethrough, and the first power feeder 31 and the second power feeder 32 are electrically connected via the diaphragm 33 .
- electrolysis proceeds without increasing the pH value of the electrolyzed hydrogen water (that is, while the water in the electrolysis chamber 30 is maintained neutral). do.
- hydrogen gas and oxygen gas are generated.
- oxygen gas is generated in the first electrode chamber 30A, and electrolyzed oxygen water in which the oxygen gas is dissolved is produced.
- hydrogen gas is generated in the second electrode chamber 30B, and electrolyzed hydrogen water in which the hydrogen gas is dissolved is produced.
- oxygen gas is generated in the second electrode chamber 30B, and electrolyzed oxygen water in which the oxygen gas is dissolved is produced.
- the electrolytic cell 4 includes a first power feeder 41 and a second power feeder 42 arranged facing each other in an electrolysis chamber 40 for electrolyzing water, and the electrolysis chamber 40 is placed on the first power feeder 41 side. It has a diaphragm 43 that separates the first electrode chamber 40A and the second electrode chamber 40B on the second feeder 42 side.
- One of the first power feeder 41 and the second power feeder 42 is applied as an anode power feeder, and the other is applied as a cathode power feeder. Water is supplied to both the first electrode chamber 40A and the second electrode chamber 40B of the electrolysis chamber 40, and a DC voltage is applied to the first power feeder 41 and the second power feeder 42, whereby water is generated in the electrolysis chamber 40. electrolysis occurs.
- the diaphragm 43 is composed of, for example, a polytetrafluoroethylene (PTFE) hydrophilic film.
- a metal plate such as titanium, for example, is applied to the first power feeder 41 and the second power feeder 42 that are arranged to face each other with the diaphragm 43 interposed therebetween.
- the first power feeder 41 and the second power feeder 42 are arranged at positions separated from the diaphragm 43 .
- electrolysis proceeds while the pH value of the electrolyzed hydrogen water increases, that is, as the alkalinity of the water in the cathode chamber increases.
- first power feeder 41 When the first power feeder 41 is applied as an anode power feeder, oxygen gas is generated in the first electrode chamber 40A, and electrolyzed oxygen water in which the oxygen gas is dissolved is produced. On the other hand, hydrogen gas is generated in the second electrode chamber 40B, and electrolyzed hydrogen water in which the hydrogen gas is dissolved is produced.
- first power feeder 41 When the first power feeder 41 is applied as a cathode power feeder, hydrogen gas is generated in the first electrode chamber 40A, and electrolyzed hydrogen water in which the hydrogen gas is dissolved is produced.
- oxygen gas is generated in the second electrode chamber 40B, and electrolyzed oxygen water in which the oxygen gas is dissolved is produced.
- the electrolyzed water generator 1 has a water supply channel 20 for supplying water to be electrolyzed to the electrolysis chambers 30 and 40, and water discharge channels 61 and 62 for discharging the electrolyzed water from the electrolysis chambers 30 and 40. ing.
- Raw water is supplied to the electrolyzed water generator 1 from the water supply channel 20 .
- raw water tap water is generally used, but other sources such as well water and ground water can also be used.
- a water purification cartridge or the like for purifying raw water is appropriately provided in the water supply path 20 .
- the water supply channel 20 is branched into a water supply channel 21 and a water supply channel 22 .
- the water supply path 21 is connected to the lower part of the first electrode chamber 30A.
- the water supply path 22 is connected to the lower end of the second pole chamber 30B. The water flowing into the water supply passage 20 passes through the water supply passages 21 and 22 and flows into the first electrode chamber 30A and the second electrode chamber 30B.
- a flow rate adjustment valve 23 is provided in the water supply paths 21 and 22, and this flow rate adjustment valve 23 adjusts the amount of water flowing through the water supply paths 21 and 22. Then, the amount of water flowing into the first electrode chamber 30A and the second electrode chamber 30B is adjusted by the flow control valve 23 .
- the water discharge passage 61 is connected to the upper end of the first pole chamber 40A, whereby the water flowing out of the first pole chamber 40A flows into the water discharge passage 61. Also, the water discharge passage 62 is connected to the upper end portion of the second pole chamber 40B, whereby the water flowing out of the second pole chamber 40B flows into the water discharge passage 62. As shown in FIG.
- a flow passage switching valve 63 is provided between the first pole chamber 40A and the second pole chamber 40B and the water discharge passages 61 and 62.
- the connection between the second pole chamber 40B and the water discharge passages 61, 62 is selectively switched.
- the electrolysis currents supplied to the feeders 31, 32 and the feeders 41, 42 are controlled by a controller (not shown).
- the control unit controls each unit such as the power feeders 31 and 32 and the power feeders 41 and 42 .
- the control unit has, for example, a CPU (Central Processing Unit) that executes various kinds of arithmetic processing, information processing, etc., a memory that stores programs that control the operation of the CPU, and various kinds of information.
- the controller controls the polarities of the first power feeders 31, 41 and the second power feeders 32, 42, for example.
- first power feeders 31, 41 and the second power feeders 32, 42 by mutually changing the polarities of the first power feeders 31, 41 and the second power feeders 32, 42, desired electrolyzed water out of electrolyzed hydrogen water or electrolyzed oxygen water is discharged from the discharge channel 61, and unnecessary Electrolyzed water can be discharged from the water discharge channel 62 .
- the time during which the first power feeders 31 and 41 and the second power feeders 32 and 42 function as anode power feeders or cathode power feeders can be made uniform, and scale adhesion in the electrolytic chambers 30 and 40 can be suppressed.
- the first electrode chamber 30A of the electrolysis chamber 30 and the first electrode chamber 40A of the electrolysis chamber 40 are communicated in series by the first water channel 51.
- the second electrode chamber 30B of the electrolysis chamber 30 and the second electrode chamber 40B of the electrolysis chamber 40 are communicated in series by the second water passage 52 .
- hydrogen water for suppressing alcoholic liver injury that is mixed with ethanol is used.
- a solution in which hydrogen is dissolved may also be used.
- a membrane module that includes a sleeve to which hydrogen gas is supplied and a hollow fiber that is arranged inside the sleeve and has a plurality of holes formed therein, the holes formed in the hollow fibers Through bubbling, hydrogen gas is brought into direct contact with an ethanol solution to produce an ethanol solution in which hydrogen is dissolved.
- electrolyzed hydrogen water of levels 1 to 4 with different dissolved hydrogen concentrations was produced under the conditions of a temperature of 22 ° C. and a flow rate of 1.5 L / min. and obtained purified water filtered by micro carbon.
- the pH and dissolved hydrogen concentration of the electrolyzed hydrogen water at each level above were measured.
- the pH was measured using a pH meter (manufactured by HORIBA, trade name: LAQUA act D-71), and the dissolved hydrogen concentration was measured using a dissolved hydrogen meter DH-35A (manufactured by Toa DKK). rice field.
- Level 1 800 ppb, pH 7 Level 2: 860 ppb, pH 8 Level 3: 1120 ppb, pH 9 Level 4: 1320 ppb, pH 10
- DMEM Dulbecco's Modified Eagle Medium
- the dissolved hydrogen concentration in each level of electrolytic hydrogen water medium is as follows.
- HepG2 cell line which is a human liver cancer-derived cell, was obtained from RIKEN BioResource Research Center (RIKEN BRC), 10% Fetal Bovine Serum (FBS) (Sigma-Aldrich, F7524), and 1% penicillin-streptomycin (Fujifilm Co., Ltd.).
- the cells were cultured in DMEM containing Kojunyaku, trade name: 168-23191) at 37° C. and 5% CO 2 .
- HepG2 cells were seeded at 5.0 ⁇ 10 4 cells/24-well plate, pre-cultured for 24 hours, and treated with 0-8% ethanol for 24 hours. Then, the cells are detached and suspended using trypsin, and the viable cell count is measured using a staining method (trypan blue method) using trypan blue (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd., trade name: 207-17081). (1) was used to calculate the cell viability of HepG2 cells in ethanol treatment. Table 1 shows the above results.
- HepG2 cells were seeded at 5.0 ⁇ 10 4 cells/24well plate, and after pre-culture for 24 hours, purified water, electrolyzed hydrogen water medium at levels 1 to 4, purified water containing 4% ethanol, and 4% ethanol were added. It was treated for 24 hours with an electrolyzed hydrogen water medium containing levels 1 to 4. Then, the cells are detached and suspended using trypsin, and the viable cell count is measured using a staining method (trypan blue method) using trypan blue (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd., trade name: 207-17081). Equation (2) was used to calculate the cell viability of HepG2 cells in ethanol treatment. Tables 2 and 3 show the above results.
- HepG2 cells were seeded at 5.0 ⁇ 10 4 cells/24well plate, pre-cultured for 24 hours, purified water containing 0-8% ethanol, and level 4 electrolyzed hydrogen water containing 0-8% ethanol. The medium was treated for 24 hours. Then, the cells are detached and suspended using trypsin, and the viable cell count is measured using a staining method (trypan blue method) using trypan blue (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd., trade name: 207-17081). (3) was used to calculate the cell viability of HepG2 cells in ethanol treatment. Tables 4 and 5 show the above results.
- HepG2 cells were seeded at 5.0 ⁇ 10 4 cells/24well plate, pre-cultured for 24 hours, purified water, level 4 electrolyzed hydrogen water culture medium, and level 4 electrolyzed hydrogen water that was autoclaved (AC) twice.
- the cell viability when treated with a hydrogen water medium having a dissolved hydrogen concentration of 1040 ppb, the cell viability was about the same as when treated with an electrolytic hydrogen water medium containing 4% ethanol at level 4, and the cell viability was generated by electrolytic treatment. It can be seen that even if it is not hydrogen water, it has the effect of reducing the toxicity of ethanol to liver cells, like electrolyzed hydrogen water.
- HepG2 cells were seeded at 2.0 ⁇ 10 5 cells/6well plate, precultured for 24 hours, purified water containing 4% ethanol, level 4 electrolytic hydrogen water medium containing 4% ethanol, 4% A level 4 electrolytic hydrogen water medium (AC) containing ethanol and subjected to two autoclave (AC) treatments, and a hydrogen water medium containing 4% ethanol and having a dissolved hydrogen concentration of 1040 ppb (5 x Dulbecco's Modified Eagle Medium (DMEM) diluted 5-fold with hydrogen water having a dissolved hydrogen concentration of 1300 ppb) was treated for 24 hours.
- DMEM Dulbecco's Modified Eagle Medium
- HepG2 cells were seeded at 1.0 ⁇ 10 6 cells/10-cm dish and precultured for 24 hours.
- DMEM 5x Dulbecco's Modified Eagle Medium
- HepG2 cells were seeded at 2.0 ⁇ 10 5 cells/6well plate, pre-cultured for 24 hours, purified water, level 4 electrolyzed hydrogen water medium, and level 4 electrolyzed hydrogen water that was autoclaved (AC) twice.
- a medium (AC) a hydrogen water medium with a dissolved hydrogen concentration of 1040 ppb (the above 5x Dulbecco's Modified Eagle Medium (DMEM) diluted 5 times with hydrogen water with a dissolved hydrogen concentration of 1300 ppb), and these The cells were treated with water or medium containing 4% ethanol for 24 hours.
- HepG2 cells were seeded at 2.0 ⁇ 10 5 cells/6well plate, pre-cultured for 24 hours, purified water, level 4 electrolyzed hydrogen water medium, and level 4 electrolyzed hydrogen water that was autoclaved (AC) twice.
- a medium (AC) a hydrogen water medium with a dissolved hydrogen concentration of 1040 ppb (the above 5x Dulbecco's Modified Eagle Medium (DMEM) diluted 5 times with hydrogen water with a dissolved hydrogen concentration of 1300 ppb), and these The cells were treated with water or medium containing 4% ethanol for 24 hours.
- HepG2 cells were seeded at 5.0 ⁇ 10 4 cells/24well plate, pre-cultured for 24 hours, purified water, level 4 electrolyzed hydrogen water medium, and level 4 electrolyzed hydrogen water that was autoclaved (AC) twice.
- a medium (AC) a hydrogen water medium with a dissolved hydrogen concentration of 1040 ppb (the above 5x Dulbecco's Modified Eagle Medium (DMEM) diluted 5 times with hydrogen water with a dissolved hydrogen concentration of 1300 ppb), and these The cells were treated with water or medium containing 1 mM acetaldehyde for 24 hours.
- Cell viability (number of viable cells in aldehyde-treated group/number of viable cells in aldehyde-untreated group) x 100 (4)
- level 4 electrolytic hydrogen water medium containing 1 mM aldehyde As shown in Table 13, when treated with a level 4 electrolytic hydrogen water medium containing 1 mM aldehyde, the cell survival rate is improved compared to when treated with purified water containing 1 mM aldehyde. It can be seen that level 4 electrolyzed hydrogen water has the effect of reducing the toxicity of aldehyde to liver cells.
- the cell survival rate is improved compared to the case of treatment with purified water containing 1 mM aldehyde. It can be seen that it has the effect of reducing the toxicity of aldehydes to
- Hydrogen water of 7000 ppb was generated using a hydrogen water generation kit (trade name: TRIM SEVEN WATER manufactured by Nihon Trim Co., Ltd.).
- the hydrogen generating agent attached to the kit that had been immersed in purified water for 5 seconds was placed in a dedicated capsule attached to the kit, and then placed in a PET bottle attached to the kit filled with purified water, sealed, and left to stand for 5 minutes.
- the PET bottle was shaken for 30 seconds and allowed to stand at 4° C. for 24 hours.
- the PET bottle was shaken for 30 seconds to obtain 7000 ppb hydrogen water.
- 1300 ppb hydrogen water was obtained by diluting 7000 ppb hydrogen water with purified water.
- HepG2 cells were seeded in 5.0 ⁇ 10 4 cells/24well plate, and after preculture for 24 hours, purified water, a hydrogen water medium with a dissolved hydrogen concentration of 1040 ppb, a hydrogen water medium with a dissolved hydrogen concentration of 5600 ppb (The above-mentioned 5x Dulbecco's Modified Eagle Medium (DMEM) was diluted 5-fold with the above-mentioned hydrogen water having a dissolved hydrogen concentration of 1300 ppb or 7000 ppb), and each of these waters or media contained 4% ethanol. and treated for 24 hours.
- DMEM Dulbecco's Modified Eagle Medium
- Equation (2) was used to calculate the cell viability of HepG2 cells in ethanol treatment. Tables 14 and 15 show the above results.
- electrolytic cell 1 electrolyzed water generator 3 electrolytic cell 4 electrolytic cell 20-22 water supply path 30 electrolytic chamber 30A first electrode chamber 30B second electrode chamber 31 first feeder 32 second feeder 33 diaphragm 40 electrolytic chamber 40A first electrode chamber 40B Second electrode chamber 41 First power feeder 42 Second power feeder 43 Diaphragm 61 Water discharge channel 62 Water discharge channel
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Abstract
Description
電解水生成装置(日本トリム社製、商品名:TRIMION GRACE)を用いて、温度が22℃、流速が1.5L/minの条件で、溶存水素濃度の異なるレベル1~4の電解水素水を得るとともに、マイクロカーボンによってろ過された浄水を得た。
レベル2:860ppb、pH8
レベル3:1120ppb、pH9
レベル4:1320ppb、pH10
粉体より調整した5×ダルベッコ変法イーグル培地(DMEM)を、上述の浄水または各レベルの電解水素水で5倍に希釈し、電解水素水の処理用培地(すなわち、20%の5×DMEMと、80%の電解水素水または浄水を混和した水素水混合物である処理用培地)として用いた。
レベル2:860ppb×4/5=688ppb
レベル3:1120ppb×4/5=896ppb
レベル4:1320ppb×4/5=1056ppb
ヒト肝癌由来細胞であるHepG2細胞株を理研バイオリソース研究センター(RIKEN BRC)より入手し、10%のFetal Bovine Serum(FBS)(Sigma-Aldrich、F7524)、および1%のペニシリン-ストレプトマイシン(富士フイルム和光純薬社製、商品名:168-23191)を含むDMEMにて37℃、5%CO2の条件下で培養した。
HepG2細胞を5.0×104cells/24well plateで播種し、24時間前培養後、0~8%のエタノールで24時間処理した。その後、トリプシンを用いて細胞を剥離・懸濁し、トリパンブルー(富士フイルム和光純薬社製、商品名:207-17081)による染色法(トリパンブルー法)を用いて生細胞数を計測し、以下の式(1)を使用して、エタノール処理におけるHepG2細胞の細胞生存率を算出した。以上の結果を表1に示す。
細胞生存率=(各エタノール濃度の生細胞数/エタノール濃度が0%の場合の生細胞数)×100 (1)
HepG2細胞を5.0×104cells/24well plateで播種し、24時間前培養後、浄水、レベル1~4の電解水素水培地、4%のエタノールを含有する浄水、及び4%のエタノールを含有するレベル1~4の電解水素水培地で24時間処理した。その後、トリプシンを用いて細胞を剥離・懸濁し、トリパンブルー(富士フイルム和光純薬社製、商品名:207-17081)による染色法(トリパンブルー法)を用いて生細胞数を計測し、以下の式(2)を使用して、エタノール処理におけるHepG2細胞の細胞生存率を算出した。以上の結果を表2、表3に示す。
細胞生存率=(エタノール処理群の生細胞数/エタノール未処理群の生細胞数)×100 (2)
HepG2細胞を5.0×104cells/24well plateで播種し、24時間前培養後、0~8%のエタノールを含有する浄水、及び0~8%のエタノールを含有するレベル4の電解水素水培地で24時間処理した。その後、トリプシンを用いて細胞を剥離・懸濁し、トリパンブルー(富士フイルム和光純薬社製、商品名:207-17081)による染色法(トリパンブルー法)を用いて生細胞数を計測し、以下の式(3)を使用して、エタノール処理におけるHepG2細胞の細胞生存率を算出した。以上の結果を表4、表5に示す。
細胞生存率=(各エタノール濃度の生細胞数/エタノール濃度が0%の場合の生細胞数)×100 (3)
HepG2細胞を5.0×104cells/24well plateで播種し、24時間前培養後、浄水、レベル4の電解水素水培地、2回のオートクレーブ(AC)処理を行ったレベル4の電解水素水培地(AC)、緩衝液(HEPES)によりpH調整(中和)したレベル4の電解水素水培地(pH)、溶存水素濃度が1040ppbである水素水培地(上述の5×ダルベッコ変法イーグル培地(DMEM)を、溶存水素濃度が1300ppbの水素水で5倍に希釈したもの)、及びこれらの水または培地の各々に4%のエタノールを含有させたもので24時間処理した。その後、トリプシンを用いて細胞を剥離・懸濁し、トリパンブルー(富士フイルム和光純薬社製、商品名:207-17081)による染色法(トリパンブルー法)を用いて生細胞数を計測し、上記の式(2)を使用して、エタノール処理におけるHepG2細胞の細胞生存率を算出した。以上の結果を表6、表7に示す。
HepG2細胞を2.0×105cells/6well plateで播種し、24時間前培養後、4%のエタノールを含有する浄水、4%のエタノールを含有するレベル4の電解水素水培地、4%のエタノールを含有し、2回のオートクレーブ(AC)処理を行ったレベル4の電解水素水培地(AC)、及び4%のエタノールを含有し、溶存水素濃度が1040ppbである水素水培地(上述の5×ダルベッコ変法イーグル培地(DMEM)を、溶存水素濃度が1300ppbの水素水で5倍に希釈したもの)で24時間処理した。
HepG2細胞を1.0×106cells/10-cm dishで播種し、24時間前培養後、4%のエタノールを含有する浄水、4%のエタノールを含有するレベル4の電解水素水培地、4%のエタノールを含有し、2回のオートクレーブ(AC)処理を行ったレベル4の電解水素水培地(AC)、及び4%のエタノールを含有し溶存水素濃度が1040ppbである水素水培地(上述の5×ダルベッコ変法イーグル培地(DMEM)を、溶存水素濃度が1300ppbの水素水で5倍に希釈したもの)で24時間処理した。
HepG2細胞を2.0×105cells/6well plateで播種し、24時間前培養後、浄水、レベル4の電解水素水培地、2回のオートクレーブ(AC)処理を行ったレベル4の電解水素水培地(AC)、溶存水素濃度が1040ppbである水素水培地(上述の5×ダルベッコ変法イーグル培地(DMEM)を、溶存水素濃度が1300ppbの水素水で5倍に希釈したもの)、及びこれらの水または培地の各々に4%のエタノールを含有させたもので24時間処理した。
HepG2細胞を2.0×105cells/6well plateで播種し、24時間前培養後、浄水、レベル4の電解水素水培地、2回のオートクレーブ(AC)処理を行ったレベル4の電解水素水培地(AC)、溶存水素濃度が1040ppbである水素水培地(上述の5×ダルベッコ変法イーグル培地(DMEM)を、溶存水素濃度が1300ppbの水素水で5倍に希釈したもの)、及びこれらの水または培地の各々に4%のエタノールを含有させたもので24時間処理した。
HepG2細胞を5.0×104cells/24well plateで播種し、24時間前培養後、浄水、レベル4の電解水素水培地、2回のオートクレーブ(AC)処理を行ったレベル4の電解水素水培地(AC)、溶存水素濃度が1040ppbである水素水培地(上述の5×ダルベッコ変法イーグル培地(DMEM)を、溶存水素濃度が1300ppbの水素水で5倍に希釈したもの)、及びこれらの水または培地の各々に1mMのアセトアルデヒドを含有させたもので24時間処理した。その後、トリプシンを用いて細胞を剥離・懸濁し、トリパンブルー(富士フイルム和光純薬社製、商品名:207-17081)による染色法(トリパンブルー法)を用いて生細胞数を計測し、以下の式(4)を使用して、アルデヒド処理におけるHepG2細胞の細胞生存率を算出した。以上の結果を表12、表13に示す。
細胞生存率=(アルデヒド処理群の生細胞数/アルデヒド未処理群の生細胞数)×100 (4)
水素水生成キット(日本トリム社製、商品名:TRIM SEVEN WATER)を用いて7000ppbの水素水を生成した。次に、浄水に5秒浸したキット付属の水素発生剤をキット付属の専用カプセルに入れた後、浄水で満たしたキット付属のペットボトルに入れて密栓し、5分静置した。その後、そのペットボトルを30秒間振盪させた後、4℃にて24時間、静置した。そして、そのペットボトルを30秒間振盪し、7000ppbの水素水を得た。なお、1300ppbの水素水は、7000ppbの水素水を浄水で希釈することにより得た。
3 電解槽
4 電解槽
20~22 給水路
30 電解室
30A 第1極室
30B 第2極室
31 第1給電体
32 第2給電体
33 隔膜
40 電解室
40A 第1極室
40B 第2極室
41 第1給電体
42 第2給電体
43 隔膜
61 吐水路
62 吐水路
Claims (4)
- エタノールと混和されるアルコール性肝障害抑制用の水素水であって、
溶存水素濃度が550~5600ppbであり、
前記エタノールと混和した場合の前記水素水におけるエタノールの濃度が1~4%であることを特徴とするアルコール性肝障害抑制用の水素水。 - 前記溶存水素濃度が1100~1300ppbであることを特徴とする請求項1に記載のアルコール性肝障害抑制用の水素水。
- 電解水素水であることを特徴とする請求項1または請求項2に記載のアルコール性肝障害抑制用の水素水。
- 溶存水素濃度が550~5600ppbであるアルコール性肝障害抑制用の水素水を、該水素水におけるエタノールの濃度が1~4%となるように、エタノールと混和することを特徴とするエタノールの処理方法。
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JPH06116144A (ja) | 1992-10-07 | 1994-04-26 | Shuichi Kimura | アルコール性肝障害予防用組成物 |
JPH06247876A (ja) | 1993-02-22 | 1994-09-06 | Snow Brand Milk Prod Co Ltd | アルコール性肝障害軽減剤 |
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US20240050465A1 (en) | 2024-02-15 |
JP2022126128A (ja) | 2022-08-30 |
CN116847856A (zh) | 2023-10-03 |
EP4282422A1 (en) | 2023-11-29 |
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