JPH07244037A - Measurement of hair growing effect - Google Patents
Measurement of hair growing effectInfo
- Publication number
- JPH07244037A JPH07244037A JP3248594A JP3248594A JPH07244037A JP H07244037 A JPH07244037 A JP H07244037A JP 3248594 A JP3248594 A JP 3248594A JP 3248594 A JP3248594 A JP 3248594A JP H07244037 A JPH07244037 A JP H07244037A
- Authority
- JP
- Japan
- Prior art keywords
- hair
- mrna
- g6pdh
- amt
- developing
- 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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Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は、育毛成分が有する育毛
効果を迅速かつ高精度に測定する方法に関する。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for quickly and highly accurately measuring the hair-growth effect of hair-growth ingredients.
【０００２】[0002]
【従来の技術】育毛成分として、生薬、有機化合物、無
機化合物等の種々の有効成分が知られているが、これら
の育毛効果を評価する方法としては、一般にはマウスや
人を使用した評価方法が知られている。2. Description of the Related Art Various active ingredients such as crude drugs, organic compounds and inorganic compounds are known as hair-growth ingredients. As a method for evaluating the hair-growth effect, generally, an evaluation method using a mouse or human is used. It has been known.
【０００３】具体的には、マウス等の動物の体毛を除毛
し、その部位に育毛成分あるいはこれらを含む育毛剤を
一定量塗布し、体毛の伸びを目視或いは色差計により評
価する方法や、脱毛症、薄毛症のパネラーに育毛剤を連
続使用して貰い、アンケート結果や目視ないしは写真判
定により評価する方法が一般に採用されている。Specifically, a method of removing hair from an animal such as a mouse, applying a certain amount of a hair-growth component or a hair-growth agent containing these to the area, and evaluating the growth of the hair visually or with a color difference meter, A method of continuously using a hair-growth agent for alopecia and thinning hair panelists, and evaluating by a questionnaire result or visual or photographic judgment is generally adopted.
【０００４】ところで、グルコース−６−リン酸脱水素
酵素（以下、「Ｇ６ＰＤＨ」と略す）は、グルコース−
６−リン酸を基質とし、ＮＡＤＰ+の存在下で、６−ホ
スホノ−γ−ラクトンとＮＡＤＰＨを生じる反応を触媒
する酵素である。この酵素による反応は、動植物の代謝
上、解糖系とペントース−リン酸経路の分岐点に位置
し、生体の代謝調節上重要な意義を有すると共に、各種
の合成反応におけるＮＡＤＰＨの供給にあずかることが
知られている。さらに、毛球部におけるＧ６ＰＤＨ活性
と毛幹の太さには相関関係が認められることが本発明者
らによって知見されている（粧技誌第２５巻第２号１３
４〜１３９頁）が、この知見を育毛成分の効果の評価に
応用する可能性を示唆するものではなかった。By the way, glucose-6-phosphate dehydrogenase (hereinafter abbreviated as "G6PDH") is glucose-
It is an enzyme that catalyzes a reaction that produces 6-phosphono-γ-lactone and NADPH in the presence of NADP + using 6-phosphate as a substrate. The reaction by this enzyme is located at the branch point of glycolysis and pentose-phosphate pathway in the metabolism of animals and plants, has important significance in the regulation of metabolism in the living body, and participates in the supply of NADPH in various synthetic reactions. It has been known. Furthermore, it has been found by the present inventors that there is a correlation between the G6PDH activity in the hair bulb portion and the thickness of the hair shaft (Cosmetic Technical Journal Vol. 25, No. 2, 13).
4 to 139) did not suggest the possibility of applying this finding to the evaluation of the effect of the hair-growth component.
【０００５】[0005]
【発明が解決しようとする課題】発毛・育毛効果の評価
方法に於いては、効果を客観的且つ精度良く評価できる
ことが重要である。しかしながら、上記の評価方法では
評価に要する期間が動物を使用した場合には通常３週間
以上、パネラーによる場合は通常４ヶ月以上かかる上、
評価基準が主観的なので効果の判定精度が曖昧なものに
なりやすかった。In the method of evaluating the hair growth / hair growth effect, it is important to be able to evaluate the effect objectively and accurately. However, in the above evaluation method, the period required for the evaluation is usually 3 weeks or longer when animals are used and 4 months or longer when the panel is used.
Since the evaluation criteria are subjective, the accuracy of determining the effect tends to be ambiguous.
【０００６】本発明は、斯かる実情に鑑みてなされたも
のであって、育毛成分の効果を迅速且つ客観的に精度良
く評価できる方法を提供する事を課題とする。The present invention has been made in view of such circumstances, and an object thereof is to provide a method capable of quickly and objectively and accurately evaluating the effect of a hair-growth component.
【０００７】[0007]
【課題を解決するための手段】本発明者は、上記課題を
解決するため鋭意研究を重ねた結果、毛包のグルコース
−６−リン酸脱水素酵素の酵素活性が毛幹の太さと相関
する事を知見した。更にこの酵素に着目し研究を進めた
結果、Ｇ６ＰＤＨをコードするｍＲＮＡ（以下、「Ｇ６
ＰＤＨ−ｍＲＮＡ」という）の発現量が育毛効果と強く
相関する事を見出し、本発明を完成させたものである。Means for Solving the Problems As a result of intensive studies to solve the above problems, the present inventor has found that the enzyme activity of glucose-6-phosphate dehydrogenase in hair follicles correlates with the thickness of the hair shaft. I found things. As a result of further research focusing on this enzyme, the mRNA encoding G6PDH (hereinafter referred to as "G6PDH"
The inventors have found that the expression level of "PDH-mRNA" strongly correlates with the hair-growth effect and completed the present invention.
【０００８】すなわち本発明は、実験動物の皮膚又は毛
包由来の培養細胞に試料を投与し、前記皮膚中の毛包又
は培養細胞中のグルコース−６−リン酸脱水素酵素をコ
ードするｍＲＮＡの発現量を測定することにより、前記
試料の育毛効果を測定する方法である。That is, the present invention is to administer a sample to cultured cells derived from the skin or hair follicles of an experimental animal, and to analyze the mRNA encoding glucose-6-phosphate dehydrogenase in the hair follicles or cultured cells in the skin. It is a method of measuring the hair growth effect of the sample by measuring the expression level.
【０００９】尚、本明細書において、「育毛」とは、毛
髪の発生促進及び成長促進を含み、いわゆる発毛、養毛
を含む概念である。以下、本発明を詳細に説明する。In the present specification, "hair growth" includes the promotion of hair development and growth, and is a concept including so-called hair growth and hair nourishment. Hereinafter, the present invention will be described in detail.
【００１０】本発明の方法により育毛成分の効果を評価
するに当たっては、まず評価対照である育毛成分をマウ
ス等の実験動物に投与し、所定時間経過後に毛包中のＧ
６ＰＤＨ−ｍＲＮＡの発現量を測定し、育毛成分を投与
しない場合のＧ６ＰＤＨ−ｍＲＮＡの発現量と比較す
る。Ｇ６ＰＤＨ−ｍＲＮＡの発現量は、Ｇ６ＰＤＨ−ｍ
ＲＮＡを検出することにより、毛包から調製したＲＮＡ
中におけるＧ６ＰＤＨ−ｍＲＮＡの相対量として、ある
いは一定量の毛包中のＧ６ＰＤＨ−ｍＲＮＡの絶対量と
して定量することによって、測定することができる。In evaluating the effect of the hair-growth component by the method of the present invention, first, the hair-growth component, which is an evaluation control, is administered to an experimental animal such as a mouse, and after a lapse of a predetermined time, G in the hair follicle is lapsed.
The expression level of 6PDH-mRNA is measured and compared with the expression level of G6PDH-mRNA when the hair growth component is not administered. The expression level of G6PDH-mRNA was G6PDH-m.
RNA prepared from hair follicles by detecting RNA
It can be measured by quantifying it as the relative amount of G6PDH-mRNA in the sample, or as the absolute amount of G6PDH-mRNA in a fixed amount of hair follicles.
【００１１】この際、ＲＮＡは毛包全体から調製する必
要はなく、毛母細胞等の毛球部、あるいは毛乳頭等、毛
包の一部でもよい。また、毛包以外の細胞を含んでいて
もよく、皮膚組織全体等からＲＮＡを調製してもよい。
さらに、実験動物を用いる代わりに毛乳頭細胞等の毛包
由来の培養細胞を用い、育毛成分存在下で培養した細胞
中のＧ６ＰＤＨ−ｍＲＮＡ含量を測定してもよい。At this time, RNA does not have to be prepared from the entire hair follicle, and may be a part of the hair follicle such as a hair bulb part such as a hair mother cell or a hair papilla. Further, cells other than hair follicles may be contained, and RNA may be prepared from the whole skin tissue or the like.
Furthermore, instead of using experimental animals, cultured cells derived from hair follicles such as dermal papilla cells may be used to measure the G6PDH-mRNA content in the cells cultured in the presence of the hair-growth component.
【００１２】毛包もしくはその一部を含む組織又は培養
細胞からＲＮＡを調製するには、フェノール法やチオシ
アン酸グアニジン法など、通常ＲＮＡの調製に用いられ
ている方法（例えば、Molecular Cloning second editi
on（Cold Spring Harbor Press (1989)Maniatis et.a
l.）参照）で行えばよい。To prepare RNA from tissues or cultured cells containing hair follicles or a part thereof, methods usually used for RNA preparation such as phenol method and guanidine thiocyanate method (for example, Molecular Cloning second editi
on (Cold Spring Harbor Press (1989) Maniatis et.a
l.))).
【００１３】ＲＮＡ中のＧ６ＰＤＨ−ｍＲＮＡの検出
は、Ｇ６ＰＤＨ−ｍＲＮＡに対して水素結合によってハ
イブリダイズするオリゴヌクレオチドプローブを用いた
ハイブリダイゼーション実験、例えばドットブロットハ
イブリダイゼーションあるいはノーザンハイブリダイゼ
ーション等によって検出することができる。具体的に
は、所定量のＲＮＡ溶液をナイロンメンブラン等に滴下
するなどしてメンブラン上に固定し、あるいはアガロー
スゲル電気泳動によってＲＮＡを分画した後、ゲルから
ナイロンメンブレン等にＲＮＡを転写してメンブレン上
にＲＮＡを固定し、メンブレンをプローブ溶液に浸漬し
てインキュベートすることにより、メンブレン上のＧ６
ＰＤＨ−ｍＲＮＡにハイブリダイズするプローブを検出
することにより、Ｇ６ＰＤＨ−ｍＲＮＡを検出すること
ができる。The detection of G6PDH-mRNA in RNA can be carried out by a hybridization experiment using an oligonucleotide probe which hybridizes to G6PDH-mRNA by hydrogen bonding, such as dot blot hybridization or Northern hybridization. it can. Specifically, a predetermined amount of RNA solution is dropped onto a nylon membrane or the like to fix it on the membrane, or after fractionating RNA by agarose gel electrophoresis, RNA is transferred from the gel to a nylon membrane or the like. G6 on the membrane is fixed by immobilizing RNA on the membrane, immersing the membrane in the probe solution and incubating.
G6PDH-mRNA can be detected by detecting a probe that hybridizes to PDH-mRNA.
【００１４】プローブは、放射性標識したプローブを用
いたオートラジオグラフィーによって検出、定量するこ
とができる。また、プローブをビオチン標識し、ビオチ
ン結合能を有するアビジンと酵素とのコンジュゲート、
この酵素により発色する色素と順次反応させ、発色量か
らＧ６ＰＤＨ−ｍＲＮＡにハイブリダイズしたプローブ
の量を定量することもできる。The probe can be detected and quantified by autoradiography using a radiolabeled probe. In addition, the probe is labeled with biotin, and a conjugate of avidin having a biotin-binding ability and an enzyme,
The amount of the probe hybridized to G6PDH-mRNA can be quantified from the amount of color development by sequentially reacting with a dye that develops color with this enzyme.
【００１５】プローブは、Ｇ６ＰＤＨの遺伝子のヌクレ
オチド配列が知られている（Ho, Y., Howard, A. J. et
al., Nucleic Acids Reserch 16, 7746 (1988)）の
で、その一部の配列に相補的な配列を有するオリゴヌク
レオチドを合成することにより得られる。プローブの具
体的な配列としては、配列表配列番号１に示す配列が例
示できる。As the probe, the nucleotide sequence of the G6PDH gene is known (Ho, Y., Howard, AJ et.
al., Nucleic Acids Reserch 16, 7746 (1988)), and thus can be obtained by synthesizing an oligonucleotide having a sequence complementary to a partial sequence thereof. As a specific sequence of the probe, the sequence shown in SEQ ID NO: 1 in Sequence Listing can be exemplified.
【００１６】また、毛包中のＧ６ＰＤＨ−ｍＲＮＡの発
現量は、ＰＣＲ（Polymerase chainreaction）法によっ
ても迅速に測定することができる。ＰＣＲ法は、二本鎖
ＤＮＡの一本鎖ＤＮＡへの熱変性、増幅を目的とする部
位の両端の配列に相当する２種類のオリゴヌクレオチド
と前記熱変性ＤＮＡとのアニーリング、前記オリゴヌク
レオチドをプライマーとしたポリメラーゼ反応、からな
る増幅サイクルを繰り返すことにより、前記ＤＮＡ配列
を指数関数的に増幅する方法である。このプライマーを
用いてＲＮＡを鋳型としてｃＤＮＡを合成し、引き続き
ＰＣＲ反応を行って増幅産物を定量することによりＧ６
ＰＤＨ−ｍＲＮＡの相対量を知ることができる。The expression level of G6PDH-mRNA in hair follicles can also be rapidly measured by the PCR (Polymerase chain reaction) method. The PCR method comprises heat denaturation of double-stranded DNA into single-stranded DNA, annealing of two types of oligonucleotides corresponding to sequences at both ends of a site intended for amplification, and the heat-denatured DNA, and using the oligonucleotide as a primer. The above-mentioned polymerase reaction is repeated to exponentially amplify the DNA sequence. G6 was synthesized by using this primer to synthesize cDNA using RNA as a template and then performing a PCR reaction to quantify the amplified product.
The relative amount of PDH-mRNA can be known.
【００１７】[0017]
【実施例】以下に、本発明を実施例によりさらに具体的
に説明する。EXAMPLES The present invention will be described in more detail below with reference to examples.
【００１８】[0018]
【実施例１】Ｍｅｓｓｅｎｇｅｒらの方法に従い、ラッ
トのヒゲより単離した毛乳頭細胞を１０％血清を含むイ
ーグルＭＥＭ培地に播種し、５％二酸化炭素、３７℃の
条件下で培養し、増殖した細胞を、１０％血清を含むイ
ーグルＭＥＭ平板培地（３５ｍｍ径）に４×１０4個づ
つ播種し、上記と同様の条件で２日間培養した後、育毛
作用を有するテスト試料として、ニコチン酸エチルエス
テルを１０μＭになるように、あるいはミノキシジルを
１０μＭとなるように培地に添加し、更に２日間培養し
た。培養終了後、培地から細胞を回収し、この細胞から
チオシアン酸グアニジン法により全ＲＮＡを抽出した。
得られたＲＮＡの一部について、分光光度計を用いて２
６０ｎｍの吸光度を測定し、ＲＮＡ量を定量した。Example 1 According to the method of Messenger et al., Hair papilla cells isolated from rat whiskers were inoculated in an Eagle MEM medium containing 10% serum, cultured under the conditions of 5% carbon dioxide and 37 ° C., and grown. 4 × 10 4 cells were inoculated on an Eagle MEM plate medium (35 mm diameter) containing 10% serum, and cultured for 2 days under the same conditions as above, and then ethyl nicotinic acid ester was used as a test sample having a hair growth effect. To 10 μM or minoxidil to 10 μM was added to the medium, and the cells were further cultured for 2 days. After completion of the culture, cells were recovered from the medium and total RNA was extracted from the cells by the guanidine thiocyanate method.
Using a spectrophotometer, 2
The amount of RNA was quantified by measuring the absorbance at 60 nm.
【００１９】上記ＲＮＡ中のＧ６ＰＤＨ−ｍＲＮＡの発
現量を調べるために、残りのＲＮＡをドットブロット装
置を用いてナイロンフィルターに吸着させ、ドットブロ
ットハイブリダイゼーションによる解析を行った。ハイ
ブリダイゼーション用のプロープとしては、既知のラッ
トＧ６ＰＤＨ遺伝子（Ho, Y., Howard, A. J. et al.,
Nucleic Acids Reserch 16, 7746 (1988)）の一部を参
考に、配列番号１に示したヌクレオチド配列を有するオ
リゴヌクレオチドを合成し、これを予めビオチン標識し
たものを使用した。In order to examine the expression level of G6PDH-mRNA in the above RNA, the remaining RNA was adsorbed on a nylon filter using a dot blot apparatus and analyzed by dot blot hybridization. A known rat G6PDH gene (Ho, Y., Howard, AJ et al.,
Nucleic Acids Research 16, 7746 (1988)) was partially referred to, and an oligonucleotide having a nucleotide sequence shown in SEQ ID NO: 1 was synthesized, and this was labeled with biotin in advance.
【００２０】ＲＮＡを固定したナイロンメンブランを、
プローブ溶液、アビジン−アルカリフォスファターゼ・
コンジュゲート溶液、発光基質Ｌｕｍｉｇｅｎ（ＳＴＲ
ＡＴＡＧＥＮＥ社）溶液に順次浸漬してインキュベート
し、洗浄した後Ｘ線フィルムに露光させた。得られた像
を、カラーイメージスキャナーを用いて画像データとし
て取り込み、コンピュータ解析（使用ソフトウェア：Im
age 1.31）により数値化してＧ６ＰＤＨのｍＲＮＡ発現
量を定量した。A nylon membrane on which RNA is fixed is
Probe solution, avidin-alkaline phosphatase
Conjugate solution, luminescent substrate Lumigen (STR
AGAGEN Co., Ltd.) solution was sequentially dipped and incubated, washed, and then exposed to an X-ray film. The obtained image is captured as image data using a color image scanner, and computer analysis (software used: Im
Age 1.31) to quantify the expression level of G6PDH mRNA.
【００２１】コントロールとして、ニコチン酸エステル
あるいはミノキシジルの替わりに生理食塩水５０μＬを
添加して培養した細胞から同様に全ＲＮＡを抽出してノ
ーザンハイブリダイゼーションを行い、画像解析により
得られた数値を１００として、上記で得られた値を相対
値に換算した。結果を表１に示す。As a control, total RNA was similarly extracted from cells cultured by adding 50 μL of physiological saline instead of nicotinic acid ester or minoxidil, and Northern hybridization was carried out. The value obtained by image analysis was set to 100. The values obtained above were converted into relative values. The results are shown in Table 1.
【００２２】[0022]
【表１】 [Table 1]
【００２３】[0023]
【実施例２】９週令のＣ３Ｈマウスを用いて１群を５匹
として３群を作り、各々の群のマウスの背部を除毛して
作成した部位に、除毛後の翌日、テスト試料として７０
％エタノールにニコチン酸エステルを１重量％濃度とな
るように添加した試料０．０５ｇ、７０％エタノールに
ミノキシジルを１重量％濃度となるように添加した試料
０．０５ｇ、あるいはコントロールとして、ニコチン酸
エステルを含まない７０％エタノール液０．０５ｇを塗
布し、７２時間後に当該部位を切り取った皮膚を、チオ
シアン酸グアニジン−フェノール混液（４Ｍチオシアン
酸グアニジン、２５ｍＭクエン酸（ｐＨ７）、０．５％
サルコシル、０．１Ｍ ２−メルカプトエタノールの混
合液、０．２Ｍ酢酸ナトリウム（ｐＨ４）、フェノール
及びクロロホルムを、１：０．１：１：０２の比率で混
合したもの）中でホモジネートした。得られた皮膚ホモ
ジネートからチオシアン酸グアニジン法により全ＲＮＡ
を抽出した。得られたＲＮＡの一部について、分光光度
計を用いて２６０ｎｍの吸光度を測定し、ＲＮＡ量を定
量した。Example 2 Using 9 week-old C3H mice, one group consisting of 5 mice was made into 3 groups, and the backs of the mice in each group were dehaired, and the test sample was prepared on the next day after depilation. As 70
% G of nicotinic acid ester added to 1% by weight ethanol, 0.05 g of minoxidil added to 70% ethanol to 1% by weight concentration, or nicotinic acid ester as a control 0.05 g of 70% ethanol solution not containing the above was applied, and after 72 hours, the skin cut off from the site was treated with a guanidine thiocyanate-phenol mixture solution (4 M guanidine thiocyanate, 25 mM citric acid (pH 7), 0.5%).
The mixture was homogenized in a mixture of sarcosyl, 0.1 M 2-mercaptoethanol, 0.2 M sodium acetate (pH 4), phenol and chloroform in a ratio of 1: 0.1: 1: 02). Total RNA from the obtained skin homogenate by the guanidine thiocyanate method
Was extracted. About a part of obtained RNA, the light absorbency of 260 nm was measured using the spectrophotometer, and RNA amount was quantified.
【００２４】上記ＲＮＡ中のＧ６ＰＤＨ−ｍＲＮＡの発
現量を調べるために、残りのＲＮＡをドットブロット装
置を用いてナイロンフィルターに吸着させ、実施例１と
同様にしてドットブロットハイブリダイゼーションを行
い、画像解析によりＧ６ＰＤＨのｍＲＮＡ発現量を定量
した。In order to examine the expression level of G6PDH-mRNA in the above RNA, the remaining RNA was adsorbed on a nylon filter using a dot blot apparatus, and dot blot hybridization was carried out in the same manner as in Example 1 for image analysis. The amount of G6PDH mRNA expression was quantified by.
【００２５】画像解析により得られたコントロールでの
数値を１００として、テスト試料で得られた値を相対値
に換算した。結果を表２に示す。The value obtained for the test sample was converted into a relative value, with the value of the control obtained by image analysis as 100. The results are shown in Table 2.
【００２６】[0026]
【表２】 [Table 2]
【００２７】[0027]
【比較例】９週令のＣ３Ｈマウスを用いて１群を５匹と
して３群を作り、これらのマウスの背部を２．０×２．
０ｃｍの広さに除毛して作成した部位に、除毛後の翌
日、テスト試料として７０％エタノールにニコチン酸エ
ステルを１重量％濃度となるように添加した試料０．０
５ｇ、７０％エタノールにミノキシジルを１重量％濃度
となるように添加した試料０．０５ｇ、あるいはコント
ロールとして、ニコチン酸エステルを含まない７０％エ
タノール液を０．０５ｇを塗布し、２０日後に当該部位
のＬ値を色差計（ミノルタ製）を用いて測定した。Ｌ値
の値が小さいほど黒色が強く毛密度が大きいことを示す
が、毛成長がよいものほど数値が小さくなり比較しにく
いので、Ｌ値の逆数を黒色増加度とし、テスト試料での
黒色増加度を、コントロールでの値を１００とした時の
相対値に換算して評価した。結果を表３に示す。[Comparative Example] C3H mice aged 9 weeks were used to make 3 groups, each group consisting of 5 mice, and the backs of these mice were 2.0 × 2.
A sample prepared by adding nicotinic acid ester to a concentration of 1% by weight in 70% ethanol as a test sample on the site prepared by removing hair to a width of 0 cm on the next day after hair removal.
5 g, 0.05 g of a sample prepared by adding minoxidil to 1% by weight of 70% ethanol, or as a control, 0.05 g of 70% ethanol solution containing no nicotinic acid ester was applied, and after 20 days, the site Was measured using a color difference meter (manufactured by Minolta). The smaller the L value is, the stronger the black color is and the higher the hair density is. However, the better the hair growth is, the smaller the value is and it is difficult to compare. Therefore, the reciprocal of the L value is defined as the black increase degree, and the black increase in the test sample is The degree was converted into a relative value when the value in the control was set to 100 and evaluated. The results are shown in Table 3.
【００２８】[0028]
【表３】 以上各実施例及び比較例の結果から明かなように、グル
コース−６−リン酸脱水素酵素のｍＲＮＡの発現量を検
出することにより、育毛効果を測定する方法は従来法に
比べ、きわめて短期間で効果の測定ができる上、写真や
目視による効果測定より客観性が高く、毛成長密度を色
差計で測定する方法と比較しても精度が高いものである
ことが証明された。[Table 3] As is clear from the results of the above Examples and Comparative Examples, the method for measuring the hair-growing effect by detecting the expression level of glucose-6-phosphate dehydrogenase mRNA is much shorter than the conventional method. It has been proved that the effect can be measured by the method, the objectivity is higher than the effect measurement by the photograph or the visual observation, and the accuracy is higher than the method of measuring the hair growth density by a color difference meter.
【００２９】[0029]
【発明の効果】グルコース−６−リン酸脱水素酵素のｍ
ＲＮＡの発現量を検出することにより、同酵素の発現量
を短期間で測定することができ、発毛・育毛・養毛効果
を従来の効果測定法に比べて短期間かつ高精度で評価す
ることができる。Effect of the Invention m of glucose-6-phosphate dehydrogenase
By detecting the expression level of RNA, the expression level of the enzyme can be measured in a short period of time, and hair growth, hair growth, and hair nourishing effects can be evaluated in a short period of time and with high accuracy as compared with conventional effect measurement methods. be able to.
【００３０】[0030]
配列番号：1 配列の長さ：22 配列の型：核酸 トポロジー：直鎖状 配列の種類：他の核酸 合成ＤＮＡ 配列 TGTAGTGGCA GGCTTTTCCA TG 22 SEQ ID NO: 1 Sequence length: 22 Sequence type: Nucleic acid Topology: Linear Sequence type: Other nucleic acid Synthetic DNA Sequence TGTAGTGGCA GGCTTTTCCA TG 22
Claims (2)
に試料を投与し、前記皮膚中の毛包又は培養細胞中のグ
ルコース−６−リン酸脱水素酵素をコードするｍＲＮＡ
の発現量を測定することにより、前記試料の育毛効果を
測定する方法。1. An mRNA encoding a glucose-6-phosphate dehydrogenase in a hair follicle or a cultured cell in the skin, which is obtained by administering a sample to a cultured cell derived from the skin or a hair follicle of an experimental animal.
A method for measuring the hair-growth effect of the sample by measuring the expression level of.
ードするｍＲＮＡの発現量を、このｍＲＮＡと、その一
部の配列に相補的な配列を有するオリゴヌクレオチドプ
ローブとのハイブリダイゼーションにより測定すること
を特徴とする請求項１記載の方法。2. The expression level of an mRNA encoding glucose-6-phosphate dehydrogenase is measured by hybridization of this mRNA with an oligonucleotide probe having a sequence complementary to a part of the sequence. The method according to claim 1, characterized in that
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP3248594A JPH07244037A (en) | 1994-03-02 | 1994-03-02 | Measurement of hair growing effect |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP3248594A JPH07244037A (en) | 1994-03-02 | 1994-03-02 | Measurement of hair growing effect |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH07244037A true JPH07244037A (en) | 1995-09-19 |
Family
ID=12360294
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP3248594A Pending JPH07244037A (en) | 1994-03-02 | 1994-03-02 | Measurement of hair growing effect |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH07244037A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH10265341A (en) * | 1997-03-26 | 1998-10-06 | Shiseido Co Ltd | Assay of hair-glowing agent |
| WO2005121374A2 (en) * | 2004-06-07 | 2005-12-22 | Oklahoma Medical Research Foundation | Molecular analysis of hair follicles for disease |
| JP2010070496A (en) * | 2008-09-18 | 2010-04-02 | Tsumura Lifescience Co Ltd | Agent for promoting production of hair and/or hair follicle reinforcing factor |
-
1994
- 1994-03-02 JP JP3248594A patent/JPH07244037A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH10265341A (en) * | 1997-03-26 | 1998-10-06 | Shiseido Co Ltd | Assay of hair-glowing agent |
| WO2005121374A2 (en) * | 2004-06-07 | 2005-12-22 | Oklahoma Medical Research Foundation | Molecular analysis of hair follicles for disease |
| WO2005121374A3 (en) * | 2004-06-07 | 2006-04-13 | Oklahoma Med Res Found | Molecular analysis of hair follicles for disease |
| JP2010070496A (en) * | 2008-09-18 | 2010-04-02 | Tsumura Lifescience Co Ltd | Agent for promoting production of hair and/or hair follicle reinforcing factor |
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