JPH1087505A - Evaluation of in vivo antioxidizing power - Google Patents
Evaluation of in vivo antioxidizing powerInfo
- Publication number
- JPH1087505A JPH1087505A JP8238750A JP23875096A JPH1087505A JP H1087505 A JPH1087505 A JP H1087505A JP 8238750 A JP8238750 A JP 8238750A JP 23875096 A JP23875096 A JP 23875096A JP H1087505 A JPH1087505 A JP H1087505A
- Authority
- JP
- Japan
- Prior art keywords
- radical
- skin
- test
- pref
- vivo
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Links
- 238000001727 in vivo Methods 0.000 title claims abstract description 11
- 238000011156 evaluation Methods 0.000 title claims abstract description 9
- 230000003064 anti-oxidating effect Effects 0.000 title abstract 4
- 238000012360 testing method Methods 0.000 claims abstract description 33
- 239000000126 substance Substances 0.000 claims abstract description 26
- 238000000034 method Methods 0.000 claims abstract description 23
- 229960002311 dithranol Drugs 0.000 claims abstract description 10
- 241001465754 Metazoa Species 0.000 claims abstract description 9
- NUZWLKWWNNJHPT-UHFFFAOYSA-N anthralin Chemical compound C1C2=CC=CC(O)=C2C(=O)C2=C1C=CC=C2O NUZWLKWWNNJHPT-UHFFFAOYSA-N 0.000 claims abstract description 8
- 230000003078 antioxidant effect Effects 0.000 claims description 24
- 239000003963 antioxidant agent Substances 0.000 claims description 18
- SJQBHPJLLIJASD-UHFFFAOYSA-N 3,3',4',5-tetrachlorosalicylanilide Chemical compound OC1=C(Cl)C=C(Cl)C=C1C(=O)NC1=CC=C(Cl)C(Cl)=C1 SJQBHPJLLIJASD-UHFFFAOYSA-N 0.000 claims description 2
- JQYCSXQTVLUHSQ-UHFFFAOYSA-N 8-ethyl-4-methylpyrano[3,2-f]indol-2-one Chemical compound CC1=CC(=O)OC2=C1C=C1C=CN(CC)C1=C2 JQYCSXQTVLUHSQ-UHFFFAOYSA-N 0.000 claims description 2
- 229960005260 amiodarone Drugs 0.000 claims description 2
- IYIKLHRQXLHMJQ-UHFFFAOYSA-N amiodarone Chemical compound CCCCC=1OC2=CC=CC=C2C=1C(=O)C1=CC(I)=C(OCCN(CC)CC)C(I)=C1 IYIKLHRQXLHMJQ-UHFFFAOYSA-N 0.000 claims description 2
- JFIOVJDNOJYLKP-UHFFFAOYSA-N bithionol Chemical compound OC1=C(Cl)C=C(Cl)C=C1SC1=CC(Cl)=CC(Cl)=C1O JFIOVJDNOJYLKP-UHFFFAOYSA-N 0.000 claims description 2
- 229960002326 bithionol Drugs 0.000 claims description 2
- FDDDEECHVMSUSB-UHFFFAOYSA-N sulfanilamide Chemical compound NC1=CC=C(S(N)(=O)=O)C=C1 FDDDEECHVMSUSB-UHFFFAOYSA-N 0.000 claims description 2
- 229940124530 sulfonamide Drugs 0.000 claims description 2
- ALYNCZNDIQEVRV-UHFFFAOYSA-N 4-aminobenzoic acid Chemical compound NC1=CC=C(C(O)=O)C=C1 ALYNCZNDIQEVRV-UHFFFAOYSA-N 0.000 claims 1
- HICFFJZGXWEIHN-UHFFFAOYSA-N 8-chloro-10-[3-(dimethylamino)propyl]-3-phenothiazinol Chemical compound C1=C(Cl)C=C2N(CCCN(C)C)C3=CC=C(O)C=C3SC2=C1 HICFFJZGXWEIHN-UHFFFAOYSA-N 0.000 claims 1
- 229960004050 aminobenzoic acid Drugs 0.000 claims 1
- 150000004032 porphyrins Chemical class 0.000 claims 1
- 238000005259 measurement Methods 0.000 abstract description 7
- 238000004435 EPR spectroscopy Methods 0.000 abstract description 2
- 230000005855 radiation Effects 0.000 abstract 1
- 239000003814 drug Substances 0.000 description 8
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 229940079593 drug Drugs 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- -1 lipid peroxide Chemical class 0.000 description 5
- 239000013558 reference substance Substances 0.000 description 5
- 229940088594 vitamin Drugs 0.000 description 4
- 229930003231 vitamin Natural products 0.000 description 4
- 239000011782 vitamin Substances 0.000 description 4
- 235000013343 vitamin Nutrition 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000019197 Superoxide Dismutase Human genes 0.000 description 3
- 108010012715 Superoxide dismutase Proteins 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 102000018832 Cytochromes Human genes 0.000 description 2
- 108010052832 Cytochromes Proteins 0.000 description 2
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 229940118019 malondialdehyde Drugs 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 description 2
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 2
- RVBUGGBMJDPOST-UHFFFAOYSA-N 2-thiobarbituric acid Chemical compound O=C1CC(=O)NC(=S)N1 RVBUGGBMJDPOST-UHFFFAOYSA-N 0.000 description 1
- 206010003210 Arteriosclerosis Diseases 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- AUNGANRZJHBGPY-UHFFFAOYSA-N D-Lyxoflavin Natural products OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-UHFFFAOYSA-N 0.000 description 1
- 244000000626 Daucus carota Species 0.000 description 1
- 235000002767 Daucus carota Nutrition 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000208688 Eucommia Species 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 201000004681 Psoriasis Diseases 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 1
- 229930003270 Vitamin B Natural products 0.000 description 1
- 229930003451 Vitamin B1 Natural products 0.000 description 1
- 229930003471 Vitamin B2 Natural products 0.000 description 1
- 108010093894 Xanthine oxidase Proteins 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- 229940026310 caffeine 50 mg Drugs 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 239000012676 herbal extract Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 208000028867 ischemia Diseases 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 239000003504 photosensitizing agent Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000000419 plant extract Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 229960002477 riboflavin Drugs 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 229960003080 taurine Drugs 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
- 235000019156 vitamin B Nutrition 0.000 description 1
- 239000011720 vitamin B Substances 0.000 description 1
- 235000010374 vitamin B1 Nutrition 0.000 description 1
- 239000011691 vitamin B1 Substances 0.000 description 1
- 235000019164 vitamin B2 Nutrition 0.000 description 1
- 239000011716 vitamin B2 Substances 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Landscapes
- Medicines Containing Plant Substances (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は、生薬、抗酸化ビタミ
ン、食物素材などの生体内での抗酸化力評価方法に関す
る。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for evaluating in vivo antioxidant power of crude drugs, antioxidant vitamins, food materials, and the like.
【０００２】[0002]
【従来技術】薬物、金属、虚血−再環流、ストレスなど
の種々の原因によって生成した活性酸素やフリーラジカ
ルは生体内の脂質、蛋白質、糖、ＤＮＡなどを攻撃し、
動脈硬化の発症や発癌、老化などを発生させることが知
られている。そのため、抗酸化物質といわれる、活性酸
素やフリーラジカルを消去する物質は生体を守る働きを
する。それら抗酸化物質の抗酸化力を測定することは、
医薬品などの処方を決定する場合などに重要となってい
る。2. Description of the Related Art Active oxygen and free radicals generated by various causes such as drugs, metals, ischemia-reperfusion, and stress attack lipids, proteins, sugars, DNAs, and the like in a living body.
It is known to cause onset of arteriosclerosis, carcinogenesis, aging, and the like. Therefore, a substance called an antioxidant that scavenges active oxygen and free radicals functions to protect the living body. Measuring the antioxidant power of those antioxidants,
This is important when deciding on prescriptions for pharmaceuticals.
【０００３】物質のIn vitroでの抗酸化力評価方法とし
て、抗酸化物質のスーパーオキサイドディスムターゼ
（SuperOxideDismutase,ＳＯＤ）様活性を測定するため
に、ヒポキサンチン−キサンチンオキシダーゼ系をラジ
カル生成系とし、チトクロムＣ法、ＮＢＴ（ニトロブル
ーテトロゾリウム）法、ＥＳＲ法（電子スピン共鳴法）
などを用いて解析する方法が知られている（特開平４−
５２３７）。しかし、抗酸化物質とされる天然植物中に
は還元剤が含まれているものが多いため、Ｏ2 -（スーパ
ーオキサイドアニオンラジカル）が存在しなくてもチト
クロムＣや、ＮＢＴを直接還元してしまう。そのため、
それらの物質の抗酸化力を正確に測定することは困難で
ある。また、これらの方法は酵素反応を利用した測定系
であるため、被験物質によっては、その成分が酵素を阻
害する可能性もある。実際、数種類の植物抽出物でＮＢ
Ｔ法を用いてＳＯＤ様活性を測定したところ、ＳＯＤ様
活性と考えられていたものが実は単に酵素を阻害してい
たに過ぎなかったという報告もある（山路ら、食品と開
発、３１巻、１号、４５頁（１９９６））。また、Invi
troでは活性が高くても、生体内での抗酸化作用には直
接結びつかない可能性もある。As a method for evaluating the antioxidant activity of a substance in vitro, a hypoxanthine-xanthine oxidase system is used as a radical-producing system to measure the superoxide dismutase (SOD) -like activity of the antioxidant, and cytochrome C Method, NBT (nitro blue tetrozolium) method, ESR method (electron spin resonance method)
A method of performing analysis using such methods is known (Japanese Unexamined Patent Publication No.
5237). However, because many of them contain a reducing agent naturally in plants that are antioxidants, O 2 - and cytochrome C without (superoxide anion radicals) is present, by direct reduction of NBT I will. for that reason,
It is difficult to accurately measure the antioxidant power of those substances. In addition, since these methods are measurement systems using an enzyme reaction, depending on the test substance, the components may inhibit the enzyme. In fact, several plant extracts have NB
When the SOD-like activity was measured using the T method, there was a report that what was thought to be the SOD-like activity was actually merely inhibiting the enzyme (Yamaji et al., Food and Development, Vol. 31, 1, p. 45 (1996)). Also, Invi
Although tro has high activity, it may not be directly linked to in vivo antioxidant activity.
【０００４】上述したような理由から、物質の生体内で
の抗酸化力の評価法が必要となる。生体内での抗酸化力
の評価法としては、四塩化炭素投与により肝臓で生成さ
れる過酸化脂質の生成をチオバルビツール酸反応生成物
（ThioBarbi-turic Acid Reaction Substance, ＴＢＡ
ＲＳ）としてマロンジアルデヒド（ＭＡＤ）に換算して
評価する方法がある。しかし、この方法は操作が煩雑な
うえ、感度もよくなく、また生体内で生成するラジカル
や活性酸素の抑制を直接評価する方法ではないため、臓
器などの組織の酸化がかなり進んだ状態でないと評価で
きないという欠点がある。また、生体内にはＳＯＤやカ
タラーゼなどの抗酸化酵素があり、これらの活性を高め
ることも経口投与後の抗酸化力の評価に含める必要があ
るが従来この点をも含めた評価法は知られていなかっ
た。For the reasons described above, a method for evaluating the antioxidant power of a substance in a living body is required. As a method for evaluating the antioxidant power in vivo, the production of lipid peroxide produced in the liver by administration of carbon tetrachloride is determined by using the thiobarbituric acid reaction product (TBA).
There is a method in which the evaluation is made by converting to malondialdehyde (MAD) as RS. However, this method is complicated in operation, has poor sensitivity, and is not a method for directly evaluating the suppression of radicals and active oxygen generated in a living body. There is a disadvantage that it cannot be evaluated. In addition, there are antioxidant enzymes such as SOD and catalase in the living body, and enhancing their activities also needs to be included in the evaluation of the antioxidant power after oral administration. Had not been.
【０００５】[0005]
【発明が解決しようとする課題】本発明の目的は、被験
物質の経口投与後の生体内での抗酸化力を簡易に評価す
ることにある。SUMMARY OF THE INVENTION An object of the present invention is to simply evaluate the in vivo antioxidant activity of a test substance after oral administration.
【０００６】[0006]
【課題を解決するための手段】本発明者らは被験物質を
経口投与した後の生体内での抗酸化力を組織や臓器中の
過酸化脂質の生成量ではなく、ラジカル生成抑制効果と
して評価するため、生体内でのラジカル生成量を直接測
定することを検討した。ラジカルを直接測定する方法と
してはＥＳＲ法があるが、生体内で生成するラジカルを
直接測定するのは感度的に不可能である。そこで、被験
物質を経口投与したヘアレスマウスの背部皮膚に光誘起
ラジカル生成物を塗布し、紫外線照射後、光誘起ラジカ
ル生成物塗布部分の皮膚を剥離し、ＸバンドＥＳＲで測
定すると被験物質の抗酸化力の大きさによりラジカルの
生成量が抑制されることを見いだし、その知見から本発
明を完成した。Means for Solving the Problems The present inventors evaluate the antioxidant power in vivo after oral administration of a test substance as an inhibitory effect on radical production, not on the amount of lipid peroxide produced in tissues or organs. Therefore, direct measurement of the amount of radicals generated in a living body was studied. Although there is an ESR method as a method for directly measuring radicals, it is impossible to directly measure a radical generated in a living body in terms of sensitivity. Therefore, a photo-induced radical product was applied to the back skin of a hairless mouse to which the test substance was orally administered, and after irradiation with ultraviolet light, the skin where the photo-induced radical product was applied was peeled off and measured by X-band ESR. The inventors have found that the amount of radical generation is suppressed by the magnitude of the oxidizing power, and have completed the present invention based on the findings.
【０００７】すなわち、本発明は被験物質を経口投与し
た試験動物の皮膚に光誘起ラジカル生成物を塗布し、そ
の後、光照射することにより生じるラジカル生成量を測
定することを特徴とする、被験物質の生体内抗酸化力の
評価方法である。That is, the present invention is characterized in that a photo-induced radical product is applied to the skin of a test animal to which a test substance has been orally administered, and then the amount of the radical generated by irradiation with light is measured. This is a method for evaluating the in vivo antioxidant power of a.
【０００８】[0008]
【発明の実施の形態】本発明において光誘起ラジカル生
成物とは光照射によりラジカルを生成するものであり、
具体的にはアンスラリン、３,３',４',５−テトラクロ
ロサリチルアニリド、４−メチル−Ｎ−エチルピロロ
［３,２−ｇ］クマリン、７−ヒドロキシクロルプロマ
ジン、ビチオノール、フェンチクロール、アミオダロ
ン、スルファニルアミド、４−アミノ安息香酸、ポルフ
ィリンなどがあげられる。これらのうちで最も好ましい
ものとしてアンスラリンをあげることができる。アンス
ラリンとは1,8-Dihydroxy anthroneのことであり、従来
は尋常性乾癬治療剤または光増感剤として知られている
ものである。BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, a photo-induced radical product is one that generates a radical by light irradiation,
Specifically, anthralin, 3,3 ′, 4 ′, 5-tetrachlorosalicylanilide, 4-methyl-N-ethylpyrrolo [3,2-g] coumarin, 7-hydroxychloropromazine, bitionol, fenticrol, amiodarone, Sulfanilamide, 4-aminobenzoic acid, porphyrin and the like. Of these, the most preferred is anthralin. Anthralin refers to 1,8-dihydroxy anthrone, which is conventionally known as a therapeutic agent or a photosensitizer for psoriasis vulgaris.
【０００９】本発明の方法では被験物質として、生薬、
ビタミンなどの天然物、化学合成した医薬品など経口投
与可能な物質であれば評価可能であり、さらに、それら
を組み合わせた組成物についても評価することができ
る。In the method of the present invention, crude drugs,
Any substance that can be administered orally, such as a natural product such as a vitamin or a chemically synthesized drug, can be evaluated, and a composition obtained by combining them can also be evaluated.
【００１０】本発明における試験動物としては剃毛した
ラット、マウス、ウサギなどが考えられるが、特に好ま
しいものとしてヘアレスマウスがあげられる。評価に用
いる皮膚の部位としては作業の容易さの点から背部の皮
膚が好ましい。As test animals in the present invention, shaved rats, mice, rabbits and the like can be considered, and a hairless mouse is particularly preferable. The skin on the back is preferable as the skin site used for evaluation from the viewpoint of ease of operation.
【００１１】本発明で光照射とは、光誘起ラジカル生成
物がラジカル化する光線を照射することであり、具体的
には紫外線の照射が好ましい。紫外線とは３９０ｎｍ〜
２００ｎｍの波長の光である。また、ラジカル生成量の
測定は、光照射後の試験動物の皮膚を剥離し、その皮膚
を組織用ＥＳＲ測定試料管に入れ、ＸバンドＥＳＲで測
定する方法で測定することができる。また、ＬバンドＥ
ＳＲでも測定可能であるが感度的にＸバンドＥＳＲの方
が好ましい。ラジカル強度は、Ｍｎなどの内部標準物質
との比で算出することができる。In the present invention, light irradiation means irradiation with a light beam that radicalizes a photo-induced radical product, and specifically, irradiation with ultraviolet rays is preferred. UV light is 390nm ~
This is light having a wavelength of 200 nm. The amount of radical generation can be measured by exfoliating the skin of a test animal after light irradiation, placing the skin in an ESR measurement sample tube for tissue, and measuring by X-band ESR. Also, L band E
Although it can be measured by SR, X-band ESR is more preferable for sensitivity. The radical strength can be calculated from the ratio to an internal standard such as Mn.
【００１２】被験物質の抗酸化力の評価は、基準物質と
して抗酸化作用がないことが知られている物質のラジカ
ル強度を基準に、被験物質のラジカル強度の低下の度合
により評価することができる。The antioxidant activity of the test substance can be evaluated based on the radical strength of the test substance, which is known to have no antioxidant action, based on the radical strength of the test substance. .
【００１３】[0013]
【発明の効果】本発明により、生薬、抗酸化ビタミン、
食品素材などの被験物質を経口投与したときの生体内抗
酸化力を評価することが可能になったので医薬品の処方
決定などに有用である。According to the present invention, crude drugs, antioxidant vitamins,
Since it becomes possible to evaluate the antioxidant power in vivo when a test substance such as a food material is orally administered, it is useful for determining the prescription of a drug or the like.
【００１４】[0014]
【実施例】以下、実施例に基づいて本発明をさらに詳細
に説明する。Hereinafter, the present invention will be described in more detail with reference to examples.
【００１５】製造例１ 被験物質として抗酸化作用を有するとされる生薬エキス
およびビタミン類などを組み合わせた処方、すなわち、
ニンジン ６００ｍｇ、オウセイ ３００ｍｇ、クコシ
３００ｍｇ、ジャショウシ ４００ｍｇ、トシシ ２
００ｍｇ、トチュウ ２００ｍｇ、ロクジョウ ４００
ｍｇ、インヨウカク １０００ｍｇ、カイクジン １０
０ｍｇ、ブクリョウ ４００ｍｇ、ゴミシ ３００ｍ
ｇ、ハンピ２５０ｍｇ、ビタミンＢ１ ５ｍｇ、ビタミ
ンＢ２ ５ｍｇ、ビタミンＢ６５ｍｇ、タウリン ５０
０ｍｇ、カフェイン ５０ｍｇの処方を精製水で溶解、
５０ｍｌにして被験用内服液剤を得た。Production Example 1 A prescription combining a herbal extract and vitamins which are considered to have antioxidant activity as test substances,
Carrot 600mg, Dwarf 300mg, Kukoshi 300mg, Jashoushi 400mg, Toshi 2
00mg, Eucommia 200mg, Rokujo 400
mg, Inyokaku 1000mg, Kaikujin 10
0mg, Bukuryo 400mg, Garbage 300m
g, Hampi 250mg, Vitamin B1 5mg, Vitamin B2 5mg, Vitamin B 65mg, Taurine 50
0mg, Caffeine 50mg formula dissolved in purified water,
It was made up to 50 ml to obtain a test liquid.
【００１６】実施例１ 被験物質として製造例１の内服液剤を用い、基準物質と
して抗酸化作用がないとされる生理食塩水を用いた。ま
た、光誘起ラジカル生成物としてアンスラリンを用い
た。Example 1 A test substance used was the oral liquid preparation of Preparation Example 1, and a reference substance used was physiological saline, which is considered to have no antioxidant action. Anthralin was used as a photo-induced radical product.
【００１７】試験動物はヘアレスマウス（体重２０ｇ前
後）を、１群６匹ずつ用いた。As test animals, hairless mice (weighing about 20 g) were used in groups of 6 each.
【００１８】被験物質および基準物質は１０ｍｌ／ｋｇ
を１日１回経口投与し、被験物質投与群は３日間投与群
および７日間投与群の２群、基準物質投与群は３日間投
与した群で試験を行った。各群とも、最終日の投与が終
了した５分後、マウス背部皮膚に５０ｍＭのアンスラリ
ン５００μｌを塗布し、４０ｃｍの高さの紫外線殺菌灯
（１５Ｗ，ＧＬ−１５）下に３時間放置した。その後マ
ウスを頸椎脱臼にて屠殺し、マウス背部皮膚を剥離し皮
膚切片（２×０．３ｃｍ）を作成した。作成試料を組織
用ＥＳＲ測定試料管に入れ、ＸバンドＥＳＲ（ＪＥＯ
Ｌ，ＪＥＳ−ＲＥ１Ｘ）を用いて測定した。ピーク高さ
は内部標準に用いているＭｎのピーク高さとの比をとり
ラジカル強度とした。Test substance and reference substance are 10 ml / kg
Was orally administered once a day. The test substance administration group was tested in two groups, a 3-day administration group and a 7-day administration group, and the reference substance administration group was administered in a 3-day administration group. In each group, 5 minutes after the end of the administration on the last day, 500 μl of 50 mM anthralin was applied to the skin on the back of the mouse, and the mouse was allowed to stand under an ultraviolet sterilizing lamp (15 W, GL-15) having a height of 40 cm for 3 hours. Thereafter, the mouse was sacrificed by cervical dislocation, and the skin on the back of the mouse was peeled off to prepare a skin section (2 × 0.3 cm). The prepared sample is placed in a sample tube for tissue ESR measurement, and X-band ESR (JEO
L, JES-RE1X). The peak height was defined as the radical intensity by taking the ratio with the peak height of Mn used for the internal standard.
【００１９】ＥＳＲ測定条件はマイクロ波出力：５ｍ
Ｗ、マイクロ波周波数：９４００ＭＨｚ、中心磁場：３
３３．５±１０ｍＴ、掃引時間：４ｍｉｎ、変調周波
数：１００ｋＨｚ、変調幅：０．１ｍＴ、増幅率：５×
１００、時定数：０．１ｓｅｃであった。The ESR measurement conditions are as follows: microwave output: 5 m
W, microwave frequency: 9400 MHz, central magnetic field: 3
33.5 ± 10 mT, sweep time: 4 min, modulation frequency: 100 kHz, modulation width: 0.1 mT, amplification factor: 5 ×
100, time constant: 0.1 sec.
【００２０】結果 各被験物質投与群のアンスラリンラジカルの強度を基準
物質投与群に対する割合として算出した。その結果、３
日間投与群は７１．５１％、７日間投与群は６２．７４
％（各群ともｐ＜０．００５）であった。Results The anthralin radical intensity of each test substance administration group was calculated as a ratio to the reference substance administration group. As a result, 3
71.51% for the 7-day group and 62.74 for the 7-day group
% (P <0.005 for each group).
【００２１】被験物質投与群はいずれの群も基準物質投
与群に比べ有意に皮膚中アンスラリンラジカル量が減少
し、内服液剤による生体内での抗酸化効果（ラジカル生
成抑制効果）があったことを示している。In each of the test substance administration groups, the amount of anthralin radical in the skin was significantly reduced in each group as compared with the reference substance administration group, and there was an antioxidant effect (radical generation inhibitory effect) in the living body by the internal liquid preparation. Is shown.
【００２２】このことから、被験物質経口投与後の皮膚
光照射によるアンスラリンラジカルをＸバンドＥＳＲで
測定する方法が、抗酸化物質に起因する生体内での抗酸
化効果を評価する簡便で優れた方法であることを示して
いる。From this, the method of measuring anthralin radical by X-band ESR by skin light irradiation after oral administration of a test substance is simple and excellent for evaluating the antioxidant effect in vivo due to antioxidants. Method.
Claims (4)
に光誘起ラジカル生成物を塗布し、その後、光照射する
ことにより生じるラジカル生成量を測定することを特徴
とする、被験物質の生体内抗酸化力の評価方法。1. An in vivo test substance, comprising applying a photo-induced radical product to the skin of a test animal to which the test substance has been orally administered, and then measuring the amount of radical generated by irradiation with light. Evaluation method of antioxidant power.
３,３',４',５−テトラクロロサリチルアニリド、４−
メチル−Ｎ−エチルピロロ［３,２−ｇ］クマリン、７
−ヒドロキシクロルプロマジン、ビチオノール、フェン
チクロール、アミオダロン、スルファニルアミド、４−
アミノ安息香酸、ポルフィリンからなる群から選ばれる
１種または２種以上である請求項１に記載の評価方法。2. The photoinduced radical product is an anthralin,
3,3 ', 4', 5-tetrachlorosalicylanilide, 4-
Methyl-N-ethylpyrrolo [3,2-g] coumarin, 7
-Hydroxychlorpromazine, bithionol, fenticrol, amiodarone, sulfanilamide, 4-
The evaluation method according to claim 1, wherein the evaluation method is one or more selected from the group consisting of aminobenzoic acid and porphyrin.
する請求項１または２に記載の評価方法。3. The evaluation method according to claim 1, wherein the amount of generated radicals is measured by X-band ESR.
１〜３のいずれかに記載の評価方法。4. The method according to claim 1, wherein the test animal is a hairless mouse.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP8238750A JPH1087505A (en) | 1996-09-10 | 1996-09-10 | Evaluation of in vivo antioxidizing power |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP8238750A JPH1087505A (en) | 1996-09-10 | 1996-09-10 | Evaluation of in vivo antioxidizing power |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH1087505A true JPH1087505A (en) | 1998-04-07 |
Family
ID=17034710
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP8238750A Withdrawn JPH1087505A (en) | 1996-09-10 | 1996-09-10 | Evaluation of in vivo antioxidizing power |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH1087505A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106883766A (en) * | 2015-10-02 | 2017-06-23 | 优备材料有限公司 | The method of tungsten polishing slurries and polishing substrate |
| KR20220164352A (en) | 2021-06-04 | 2022-12-13 | (주)아모레퍼시픽 | Method for evaluating an efficacy of antioxidant |
-
1996
- 1996-09-10 JP JP8238750A patent/JPH1087505A/en not_active Withdrawn
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106883766A (en) * | 2015-10-02 | 2017-06-23 | 优备材料有限公司 | The method of tungsten polishing slurries and polishing substrate |
| CN106883766B (en) * | 2015-10-02 | 2021-08-27 | 优备材料有限公司 | Tungsten polishing slurry and method of polishing substrate |
| KR20220164352A (en) | 2021-06-04 | 2022-12-13 | (주)아모레퍼시픽 | Method for evaluating an efficacy of antioxidant |
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