I I 研究活動と業績
資 源 開 発 部 門
教 授 難 波 恒 雄 助教授 門 田 重 利 助教授 小 松 か つ 子
助 手 山 路 誠 一
（薬学博士）
（薬学博士） ( 4月30日まで）
（薬学博士） (
5
月1 1
町通ら，附属薬効解析センターと併任）（薬学博士）
( 1 0
月1 6
日から）本部門では中国医学をはじめとする世界各地の伝統医学や民間療法で使用される薬物の基源の究明，真 偽鑑別，活性成分の解明，およびそれらの総合的判断による品質評価法の確立，ならびに天然薬物からの 新薬の開発などを主要研究目的としている。これらに関連して，各種生薬の組織形態学的研究，本草学的 考察，成分化学的研究，薬効発現機構の解明，薬理学的研究，さらに新薬物資源開発の目的で，世界各地 の伝統医学の比較研究，民族薬物の調査などを行っている。本年度における主たる研究内容および、成果は 次の如くである。
1
）和漢薬の生薬学的研究：キク科S e r r a t u l a
属及びキツネノマゴ科S t r o b i l a n t h e s
属に由来する漢薬「升 麻」，キク科S a u s s u r e a
属Amphilaena
亜属に由来する「雪蓮花」の基源を生薬学的に解明した。また，リンドウ科
S w e r t i a
属植物の生薬学的研究の一環として，O p h e l i a
節植物に由来する中国民間薬「樟 牙菜」及び「青葉胆」の基源を明らかにした。2
）和漢薬の遺伝子解析：富山大学工学部化学生物工学科細胞工学講座との共同研究でPan
似 属 植 物 及 びその関連生薬の1 8 Sr i b o s o m a l RN A
遺伝子の塩基配列を決定し，「人参J
類生薬同定への一手法 を提示した。また，日本及び中国産C a p t
ゐ属植物についても当該遺伝子領域の塩基配列の違いを検討 した。3
）和漢薬の品質評価に関する研究：柑橘類果実に由来する生薬の品質評価を目的として，C i t r ：
叫For‑
t u n e l l a
及びP o n c i r u s
属 29種の果皮中のポリメトキシフラボン類及ぴクマリン類計 34成分の含有パ ターンを比較検討し，それらが1 4
タイプに分けられることを明らかにした。この結果から，現在日本 及び中国市場に流通している「陳皮」，「青皮」，「根実」，「籾殻J
，「桂皮」などの原植物を検討した。4）民族医薬学の調査研究：平成 4
〜
6年度に中国中医研究院中薬研究所と共同で、行った国際学術調査「仏 教医学の調査研究」のまとめとして，大乗仏教の伝播と北方系東洋医学（チベット医学，モンゴル医 学，中国医学）との関連性について考察した。5
）和漢薬の薬効発現に関する研究：薬学部衛生化学教室及び細胞資源工学部門と共同で研究している。腸内細菌による和漢薬成分の代謝研究の一環として
B a r b a l o i n
の腸内細菌による代謝について検討し た。また腸内細菌による代謝産物を定量する目的で，酵素免疫法を用いたP a e o n i m e t a b o l i t eI
の定量 法を開発した。6
）和漢薬の薬理作用に関する研究：北海道大学薬学部薬効学講座，細胞資源工学部門との共同研究で，老化促進モデルマウス（
SAM
）の空間認知機能に対する丹参の作用について検討したほか，医学部ウ イルス学講座との共同研究でB
型肝炎ウイルス表面抗原の産生抑制作用を有する伝統薬物を検索し た。また胃内細菌H e r i c o b a c t e rp y l o r i
に対する抗菌活性物質の検索の一環として，ホップのメタノー ルエキスとl u p u l o n e
に強い活性を見出した。7
）生薬の修治に関する研究：細胞資源工学部門との共同研究で，ホミカの種々の修治法の違いによる抗 侵害受容作用について検討し，さらに医学部ウイルス学講座とも共同して単純へルペスウイルス，ポリオウイルス感染症などに対する有効性を検討した。
8 ) HIV
プロテアーゼ阻害活性を有する生薬の探索：細胞資源工学部門との共同研究で，エジプトの民間 薬についてHIV
プロテアーゼ阻害活性を指標としたスクリーニンクゃを行った。9
）和漢薬の生理活性成分の研究：青積子（C e l o s i a a r g e n t e a L
.の種子）に含有きれる酸性多糖体，及ぴ 三七人参に含まれるg i n s e n o s i d eR e , Rgl
が免疫学的肝障害に対する保護作用を有することを見出し た。またプロポリス，肉荏蓉のラジカル消去作用，升麻に含まれるトリテルペノイド類の骨粗鷺活性，雲南草果 （
A l p h i n i ab l e p h a l o c a l y x K. SCHUM.
の果実）のジアリルへブタノイドによるマクロファー ジのNO
産生阻害能などについて検討した。1 0
）伝統医薬からウイルス病治療薬の検索：医学部ウイルス学講座との共同研究で，ウイルス病治療に有 効な伝統医薬を開発することを目的として単純へルペスやサイトメグロウイルス感染症に有効な抗ウ イルス活性を示す治療薬を検索したほか，これらのへルベスウイルス自然回帰発症予防効果について も検討した。1 1
）糖尿病とその合併症治療薬の開発：天然由来の抗糖尿病薬の開発の一環としてS t r e p t o z o t o c i n
の毒 性に対するプロポリスの醇臓ベータ細胞保護作用，および合併症に関与するアルドース還元酵素に対 する丹参の阻害作用について検討した。以下，本部門で今年度中に発表し原報並ぴに口頭発表の要旨を収録し，学会活動報告とする。また社会 的活動として著書ならびに一般雑誌などへの発表および一般講演の目録を付す。
原 報
A）和漢薬の生薬学的研究
( I ) Adiantum属植物の生薬学的研究（第 5
報），胞子の形態及び小葉におけるケイ素及 びカルシウムの分布様式に基づく分類 薬学雑誌，1 1 6( 2 ) ' 1 2 5 〜 1 3 7 ( 1 9 9 6 )
小松かつ子，飯田浩一，禁 少青，御影雅幸，吉沢 健史，難波恒雄
Crude d r u g s d e r i v e d from Adiantum s p e c i e s a r e u s e d a s f e b r i f u g e , a n t i d o t e , d i u r e t i c , t o n i c , e t c . Some c o m m e r c i a l s a m p l e s o f t h e s e d r u g s a r e composed o f f i n e l y c u t u l t i m a t e p i n n u l e s o n l y , w h i c h h a v e f a l s e i n d u s i a and s p o r e s . I n t h i s p a p e r , i n o r d e r t o e s t a b ‑ l i s h a c l a s s i f i c a t i o n method b a s e d o n t h e c h a r a c t e r ‑ i s t i c s o f t h e u l t i m a t e p i n n u l e s and t h e i r a t t a c h ‑ m e n t s , t h e m o r p h o l o g i c a l s t u d y u s i n g s t e r e o s c o p e and s c a n n i n g e l e c t r o n m i c r o s c o p e , and X r a y m i ‑ c r o a n a l y s i s u s i n g an e l e c t r o n p r o b e m i c r o n a n a l y z e r were c a r r i e d o u t on t h e f a l s e i n d u s i a and s p o r e s , and t h e u l t i m a t e p i n n u l e s , r e s p e c t i v e l y , o f 1 9 Adiantum s p e c i e s . The r e s u l t s showed t h a t examined a l l s p e c i e s c o u l d b e d i s t i n g u i s h e d from e a c h o t h e r by t h e f o l l o w i n g c h a r a c t e r i s t i c s : i n t h e f a l s e i n d u s i u m ,
t h e s h a p e and t h e p r e s e n c e o r a b s e n c e o f h a i r s ; i n t h e s p o r e , t h e s h a p e , t h e o r n a m e n t a t i o n , t h e r a t i o o f t h e l a e s u r a , and t h e e q u a t o r i a l d i a m e t e r ; i n t h e
X‑r a y images o f t h e u l t i m a t e p i n n u l e , t h e d i s ‑ t r i b u t i o n a l p a t t e r n s o f s i l i c o n and c a l c i u m . The d i s t r i b u t i o n a l p a t t e r n s o f s i l i c o n were d u e t o t h e p r e s e n c e o f s p i c u l a r c e l l s , h a i r s and p a p i l l a e , and c a l c i u m was p r e s e n t a s c r y s t a l s o f c a l c i u m o x a l a t e . The a v e r a g e c o n t e n t o f s i l i c o n i n t h e u l t i m a t e p i n ‑ n u l e s o f Ad
必： n
初m s p e c i e s was 1 . 9 9 % .
( I I
）漢薬「升麻」の生薬学的研究（第1
報），C i m i c i f u g a
属植物の地下部の一般的形態及 びC . s i m p l e x
の形態変異と本種に由来する「升麻」について
生薬学雑誌，
5 0( 3 ) ' 2 2 2 〜 2 3 1 ( 1 9 9 6 )
小松かつ子，李暁波，山路誠一，難波恒雄Shengma
（升麻）i sa p o p u l a r C h i n e s e c r u d e d r u g u s e d f o r t r e a t m e n t o f e x a n t h e m a , f e v e r , p i l e s and s u p p u r a t i v e i n f l a m m a t i o n . The C h i n e s e p h a r ‑ macopoeia d e s c r i b e s t h a t t h i s d r u g i s d e r i v e d from
出 er h i z o m e s o f C i m i c i f i 昭 ah e r a c l e i f o l i a ,
C.d a h u r i c a
and C . f o e t i d a o f t h e f a m i l y R a n u n c u l a c e a e . A c c o r d ‑
i n g t o o u r f i e l d r e s e a r c h , t h e c o m m e r c i a l S h e n g ‑
ma a v a i l a b l e i n C h i n e s e markets p r e s e n t e d v a r i o u s e x t e r n a l f e a t u r e s , s u g g e s t i n g d i f f e r e n t b o t a n i c a l o r i g i n s o f t h e d r u g . I n o r d e r t o e s t a b l i s h a c r i t e r i a f o r t h e i d e n t i f i c a t i o n o f t h e b o t a n i c a l o r i g i n s o f Shengma
，a c o m p a r a t i v e a n a t o m i c a l s t u d y was c a r r i e d o u t o n 9 C i m i c
併t g a s p e c i e s . I n t h i s p a p e r , r h i z o m e s o f C . s i m ρ l e x from d i f f e r e n t growing a r e a s were examined f o r t h e i r m o r p n o l o g i c a l and a n a t o m ‑ i c a l c h a r a c t e r s . I t was f o u n d t h a t f o r a c o m p a r a t i v e s t u d y , t h e b e s t p a r t s t o b e o b s e r v e d were t h e p o r ‑ t i o n s b e t w e e n two n e i g h b o r i n g stem r e s i d u e s o f r h i z o m e w i t h more t h a n 5 stem r e s i d u e s and t h e p o r t i o n o f t h e stem r e s i d u e s 2 ‑ 3 m m up from t h e i r b a s e . The d i f f e r e n c e s o b s e r v e d between t h e r h i ‑ zomes o f C . s i m ρ l e x from Japan and t h o s e from C h i n a were i n t h e f o l l o w i n g c h a r a c t e r i s t i c s : t h e s h a p e a n d d i a m e t e r o f and d i s t a n c e between s t e m r e s i d u e s , t h i c k n e s s o f c o r t e x and s h a p e o f s e c o n d a r y xylem i n r h i z o m e ; t h e d i a m e t e r and s c l e r i f i c a t i o n o f p r i m a r y r a y c e l l s i n stem r e s i d u e ; and a p p e a r ‑ a n c e o f s c l e r e i d s i n c o r t e x i n r h i z o m e and stem r e s i d u e . Moreover
，Shengma o f Shanxi P r o v . market was i d e n t i f i e d a s C . s i m ρ J
邸fromC h i n a . ( I I I ) Pharrnacognostical Studies on the
Chinese Crude Drug Xuelianhua
（雪蓮 花）andRelated Ethnornedicines (Part 1 ) , On Chinese Crude Drug Xuelianhua
and Tibetan Crude Drug Srol‑gong
Derived from S o r o s e r i s ( Cornpositae) Plants
f
J p n . B o t .
リ7 1( 5 ) ' 2 8 8 〜 2 9 9 ( 1 9 9 6 )
S E I I C H I Y A M A J I , RUI P I N G YANG, KATSUKO KOMATSU, T I A N ‑ Z H I WANG and TSUMEO NAMBA
The C h i n e s e c r u d e d r u g X u e l i a n h u a
，t h a t i s a l s o known a s Bya‑rgod s u g ‑ p a i n T i b e t a n M e d i c i n e , i s u s e d f o r r h e u m a t o i d a r t h r i t i s , m e n o x e n i a , e t c . and h a s b e e n r e p o r t e d t o b e d e r i v e d from whole p l a n t s o f g e n u s S a
制 抑r e a o f f a m i l y C o m p o s i t a e . However, o n e s a m p l e from Songpan m a r k e t , S i c h u a n P r o v . , was f o u n d t o b e d e r i v e d from S o r o s e 的 p l a n tp o s s e s ‑ s i n g l i g u l a t e f l o w e r s and m u l t i s e r i a t e p a p p i . S o r ‑ o s e r i . s p l a n t i s r e p o r t e d t o b e u s e d f o r f e v e r and r h e u m a t o i d a r t h r i t i s i n T i b e t a n M e d i c i n e u n d e r t h e name o f S r o l ‑ g o n g
.We o b t a i n e d i t i n Thimphu m a r k e t , B h u t a n . I n t h e p r e s e n t p a p e r , t o i d e n t i f y t h e c o m m e r c i a l s a m p l e s a b o v e , a c o m p a r a t i v e a n a t o m i ‑
c a l s t u d y was c a r r i e d o u t on l e a v e s , stems and u n d e r g r o u n d p a r t s o f t h r e e s p e c i e s and o n e s u b ‑ s p e c i e s o f S o r o s e r i . s growing w i l d around T i b e t . Four t a x a c o u l d be d i s t i n g u i s h e d from e a c h o t h e r by t h e f o l l o w i n g c h a r a c t e r i s t i c s : t h e number o f v a s c u ‑ l a r b u n d l e s i n m i d r i b , t h e d e g r e e o f r o u g h n e s s o f a n t i c l i n a l w a l l o f e p i d e r m a l c e l l , and t y p e o f h a i r s i n l e a f ; p r e s e n c e o r a b s e n c e o f i n t e r f a s c i c u l a r cam‑
bium and v a s c u l a r b u n d l e c a p i n s t e m , e t c . Based on t h e s e r e s u l t s , t h e b o t a n i c a l o r i g i n s o f X u e l i a n h u a
from Songpan and S r o l ‑ g o n g from Thimphu were d e t e r m i n e d t o b e t h e whole p l a n t s i n f l o w e r i n g t o f r u i t i n g o f S o r o s e 的 h o o k e 1 匂 n a and a m i x t u r e o f t h o s e o f S . g i l l i i and S . u m b r e l l a , r e s p e c t i v e l y . (N)
「花轍」及び r( . J J
楓」の生薬学的研究（第2
報） ,
Zanthoxylum
E属に由来する漢薬「花 楓」，チベット薬物rgYer‑rna
」及びアーユ ルヴェーダ薬物「Turnburu」について 生薬学雑誌，5 0( 5 ) ' 3 2 8 〜 3 4 3 ( 1 9 9 6 )
伊 藤 親 ， 小 松 か つ 子 ， 劉 玉 捧 ， 施 大 丈 ， 難 波 恒雄
C o m m e r c i a l l y o b t a i n e d C h i n e s e c r u d e drug Hua‑
j i a o s a m p l e s may be d i v i d e d i n t o two g r o u p s
a c c o r d i n g t o t h e i r morphology : t h e f i r s t g r o u p
i n c l u d e s t h o s e d e r i v e d from t h e s u b g e n u s Z a n t h o x y ‑
lum which i s t h e m a j o r i t y , a n d t h e o t h e r group
i n c l u d e s t h o s e from t h e s u b g e n u s F a g a r a . I n t h e
p r e s e n t s t u d i e s , a c o m p a r a t i v e a n a t o m i c a l s t u d y
was p e r f o r m e d on t h e p e r i c a r p s and p e d i c e l s o f 7
s p e c i e s and 3 v a r i e t i e s o f t h e s u b g e n u s Z a n t h o x y ‑
l u m . As t h e r e s u l t , t h e s e s p e c i e s were b r o a d l y
g r o u p e d i n t o 3 c a t e g o r i e s by t h e f o l l o w i n g c h a r a c ‑
t e r i s t i c s : p r e s e n c e o r a b s e n c e o f p r o j e c t i o n s on
p e r i c a r p , d e n s i t y o f l a m e l l a r formed by c u t i c l e on
p e r i c a r p and p r e s e n c e o r a b s e n c e o f o i l s a c o f
p e d i c e l . 島 ' I o r e o v e r ,e a c h s p e c i e s o f t h e 3 c a t e g o r i e s
c o u l d be d i s t i n g u i s h e d from e a c h o t h e r by d i f f e r ‑
e n c e s i n t h e t h i c k n e s s o f p e r i c a r p and e n d o c a r p ,
t h i c k n e s s o f c u t i c l e and p r e s e n c e o r a b s e n c e o f
s c l e r e i d s i n m e s o c a r p , o f c o r k c e l l s i n c o r t e x and o f
f i b e r s o u t s i d e phloem o f p e d i c e l . Z . armatum v a r .
s u b t r i f o l
必： t u m was f u r t h e r d i v i d e d i n t o two t y p e s , A
and B , a c c o r d i n g t o t h e p r e s e n c e o r a b s e n c e o f
x a n t h o x y l i n i n p e r i c a r p , and d i f f e r e n c e s i n t h e
s h a p e o f t i p margin i n t h e d e h i s c e n c e f a c e o f p e r i ‑
c a r p and i n t h e s h a p e o f l a m e l l a on t h e s u r f a c e .
Based on t h e d i f f e r e n c e s o f each s p e c i e s , commer‑
c i a l H u a j i a o s a m p l e s were shown t o be m o s t l y mature p e r i c a r p s o f Z . bungeanum o r s l i g h t l y i m ‑ mature t o immature ones o f Z . armatum v a r . s u b t r i f o l i a t u m ( t y p e A: t y p e B=2 : 1 ) and t h e s e two k i n d s were c i r c u l a t i n g e v e n l y i n t h e r e c e n t C h i n e s e m a r k e t s . The b o t a n i c a l o r i g i n o f t h e Tibetan c r u d e drug gYer ma i n t h e Nepalese market was i d e n t i f i e d a s Z . armatum and Z . bu ngean um, and t h a t o f t h e A y u r v e d i c c r u d e drug Tumburu
＇m o s t ‑ l y a s Z . armatum i n t h e N e p a l e s e markets and a s m i x t u r e s o f Z . armatum v a r . s u b t r i f o l i a t u m ( t y p e A) and Z . bungeanum i n t h e B h u t a n e s e m a r k e t . (V
）アピセラピーとしてのプロポリスと漢薬露蜂房の比較本草学的考察 薬史学雑誌，
3 1( 2 ) ' 1 8 3 〜 1 9 9 ( 1 9 9 6 )
松繁克道，門田重利，難波恒雄A comparative s t u d y o f l i t e r a t u r e regarding p r o p o l i s and Vespae N i d u s , t o g e t h e r w i t h h i s t o r i c a l e v i d e n c e and o u r i n v e s t i g a t i o n , s u p p o r t t h e b e l i e f t h a t e a s t e r n o r i e n t a l honey b e e s do n o t produce p r o p o l i s l i k e t h e w e s t e r n honey b e e . T h e r e f o r e , i t i s n o t s u r p r i s i n g t h a t p e o p l e i n t h e e a s t u s e bee h i v e s (Vespae N i d u s ) i n s t e a d o f p r o p o l i s a s a n a t u r a l m e d i c i n e w h i l e p e o p l e i n t h e west u s e p r o p o l i s . F u r t h e r c h e m i c a l and b i o l o g i c a l s t u d i e s a r e needed t o c l a r i f y and d i s t i n g u i s h p r o p o l i s and V espae N i d u s . I n a d d i t i o n , comparative b i o l o g i c a l and c h e m i c a l s t u d i e s o f w i l d bee ( w a s p ) h i v e s s h o u l d be compared w i t h Vespae N i d u s . These s t u d i e s c o u l d s u g g e s t w h e t h e r t h e c o m p o s i t i o n o f p r o p o l i s and V e s p a e N i d u s d i f f e r due t o t h e p l a n t p r o d u c t s around them o r t h a t t h e honey b e e a l t e r s t h e compo‑
s i t i o n a f t e r c o l l e c t i o n . R e g a r d i n g t h e c o m p a r a t i v e h e r b o l o g y o f p r o p o l i s and Vespae N i d u s a s d i f f e r e n t crude drugs f o r t h e same p u r p o s e , however, r e s e a r c h on p r o p o l i s and Vespae N i d u s up t o now s u g g e s t s t h a t t h e y a r e two d i f f e r e n t d r u g s .
(VI）細辛司生薬学的研究
韓国資源植物事舎誌，
9( 2 ) ' 1 8 3 〜 1 9 9 ( 1 9 9 6 )
朴 鐘 喜 ， 金 振 水 ， 鄭 愛 栄 ， 難 波 恒 雄C h i n e s e c r u d e d r u g Se S i n
（細辛）h a s been u s e d t o c u r e h e a d a c h e , cough and e x p e c t o r a n t . To t h e b o t a n i c a l o r g i n o f Se S i n " , Zhong yao z h i
（中荊志）r e p o r t e d Asarum h e t e r o t r o ρ a i d e s v a r . mandshur ‑ i c u m , A. s i e b o l d i i and A . s i e b o l d i i v a r . s e o u l e n s i s ,
and Zhong‑yao‑da ‑ c i ‑ d i a n
（中荊大辞典）r e p o r t e d Asarum h e t e r o t r o ρ a i d e s v a r . mandshuricum and A . s i e b o l d i i o f A r i s t o l o c h i a c e a e . I n K o r e a , t h i s c r u d e drug has been u s e d a s a Korean f o l k remedy f o r t o o t h a c h e and a r o m a t i c . The b o t a n i c a l o r i g i n o f Se S i n i s c o n s i d e r e d t o be Asarum s p e c i e s o f A r i s t o l o ‑ c h i a c e a e . But t h e r e has no p h a r m a c o g n o s t i c a l c o n ‑ f i r m a t i o n on i t . To c l a r i f y t h e b o t a n i c a l o r i g i n o f Se S i n
ぺwes t u d i e d on t h e a n a t o m i c a l c h a r a c t e r i s ‑ t i c s o f Asarum s p e c i e s growing w i l d l y i n Korea i . e . A. m a c u l a t u m , A . s i e b o l d i i , A. s i e b o l d i i v a r . s e o u l e n ‑ s i s , and o f Se S i n " from Korea on Korean m a r k e t . Through o u r s t u d i e s , t h e b o t a n i c a l o r i g i n o f Se S i n from Korea was proved t o b e whole p l a n t o f Asarum s i e b o l d i i and A . s i e b o l d i i v a r . s e o u l e n s i s .
（四）中蔚細辛商品菊材的基源研究 中国中蔚雑誌、，
2 1( 1 2 ) ' 7 1 2 〜 7 1 7 ( 1 9 9 6 )
察 少 青 ， 王 瑛 ， 朱 妹 ， 李 君 山 ， 楼 之 容 ， 難波恒雄，小松かつ子
放中国28省市牧集到細辛商品荊材158f分，·~現其中佼 3／~来源子中国蔚典品科，其余1/4来源子非中国蔚典品
科9
1 1 1
t分来源北細辛，在2 6
省市使用，2 5 1
：分来源子単叶 細辛，在日省区使用，完n
伶来源子隼細辛，杜衡等9
神植物。B
）和漢薬の遺伝子解析（四）
18S Ribosomal R N A Gene Sequences of Three Panax Species and the Corre‑
sponding Ginseng Drugs
B i o l . Pharm. B u l l . , 1 9 ( 1 1 ) , 1 5 3 0 〜 1 5 3 2 ( 1 9 9 6 ) HIROTOSHI F U S H I M I , KATSUKO KAMATSU, M A S A ・ HARU ! S O B E and TSUNEO N AMBA
T o t a l DNA was e x t r a c t e d from t h e f r e s h u n d e r ‑ ground p a r t s o f t h r e e Panax s e p a r a t e s p e c i e s . The 18S rRNA r e g i o n s o f e x t r a c t e d DNA were am‑
p l i f i e d by t h e p o l y m e r a s e c h a i n r e a c t i o n (PCR) and
t h e i r s e q u e n c e s were d e t e r m i n e d . I n e a c h s p e c i e s ,
t h e s e q u e n c e s were f o u n d t o be o f 1 8 0 9 b a s e p a i r s
( b p s ) b u t w i t h d i f f e r e n t gene s e q u e n c e s . D i f f e r e n t
b a s e s u b s t i t u t i o n s were o b s e r v e d a t u c l e o t i d e p o s i ‑
t i o n s 4 9 7 , 4 9 9 , 5 0 1 and 7 1 2 , The same p r o c e d u r e was
performed on commercial samples o f Ginseng
R a d i x , P a n a c i s J a p o n i c i Rhizoma and American
G i n s e n g . Each s e q u e n c e c o m p l e t e l y c o r r e s p o n d e d
w i t h t h a t o f e a c h o r i g i n a l p l a n t , namely P . g i n s e n g ,
P . j a p o n i c u s and P . q u i n q u 包 f o l i u s , r e s p e c t i v e l y . T h i s
i s t h e f i r s t t i m e t h a t 1 8 S rRNA g e n e s e q u e n c i n g on
Panax s p e c i e s was c a r r i e d o u t . P r e v i o u s l y , Ginseng d r u g s have been i d e n t i f i e d m a i b l y by t h e i r e x t e r n a l and i n t e r n a l s t r u c t u r e . Thus t h i s method w i l l be u s e f u l i n i d e n t i f y i n g G i n s e n g d r u g s a t t h e gene l e v e l .
(IX)A New Approach for the I d e n t i f i c a t i o n of a Chinese Traditional Medicine, Chuanxiong by 188 Ribosomal R N A Gene Sequences
P h y t o m e d i c i n e , 3 ( 4 ) , 3 8 7 〜 3 8 9 ( 1 9 9 6 )
HIROTOSHI F U S H I M I , KA TSUKO KOMA TSU
，お1 A S A ・ HARU ! S O B E and TSUNEO NAMBA
We r e p o r t a new method f o r t h e i d e n t i f i c a t i o n o f a p o p u l a r C h i n e s e t r a d i t i o n a l m e d i c i n e
，Chuan x i o n g by gene s e q u e n c e s . Two s a m p l e s o f Chuan‑
xiong were chosen t h a t a r e t h e rhizomes o f Cnidium o f f i c i n a l e MAKINO, from J a p a n , and L i g u s ‑ t z
切m c h u a n x i o n g Hortorum, from C h i n a , b o t h o f f a m i l y A p i a c e a e . T h e i r 1 8 S rRNA g e n e s e q u e n c e s were t a r g e t t e d b a c a u s e t h e y c o u l d be o b t a i n e d from e v e r y o r g a n s o f t h e p l a n t and h u n d r e d s o f t h e i r c o p i e s were c o n t a i n e d i n a genome. T o t a l DNAs were e x t r a c t e d from t h e f r e s h r h i z o m e s . The 1 8 S rRNA r e g i o n s o f e x t r a c t e d DNA were a m p l i f i e d by t h e polymerase c h a i n r e a c t i o n (PCR) and t h e i r s e q u e n c e s were d e t e r m i n e d . The same p r o c e d u r e was performed on t h e c r u d e d r u g s s t o r e d w i t h i n two y e a r s . 1 8 S rRNA g e n e s e q u e n c e s o f Chuanxiong
from Japan and C h i n a were f o u n d t o be o f 1 8 0 8 b a s e s and t o c o r r e s p o n d c o m p l e t e l y w i t h t h o s e o f t h e o r i g i n a l p l a n t s , C . o f f i c i n a l e and L . c h u a n x i o n g , r e s p e c t i v e l y . However, no d i f f e r e n t b a s e s u b s t i t u ‑ t i o n was o b s e r v e d between two s p e c i e s b e c a u s e o f t h e i r taxonomic s i m i l a r i t y T h u s , we s u c c e e d e d t o e x t r a c t t h e t o t a l DNA from c r u d e d r u g s which were p r o c e s s e d and s t o r e d f o r a l o n g t i m e and t o d e t e r ‑ mine t h e i r 1 8 S rRNA gene s e q u e n c e s t h a t c o r r e ‑ sponded t o t h o s e o f o r i g i n a l p l a n t s . T h i s s u g g e s t s t h a t t h e p r e s e n t a n a l y t i c a l method o f gene s e q u e n c ‑ i n g i s u s e f u l f o r i d e n t i f i c a t i o n o f t r a d i t i o n a l m e d i ‑ c i n e s .
C
）和漢薬の品質評価に関する研究(X
）柑橘類生薬の基源と品質に関する研究（第1
報），C i t r u s
属5
種の果皮の成分及び組織形 態の成熟に伴う変化生薬学雑誌，
5 0( 2 ) ' 1 1 4 〜 1 2 7 ( 1 9 9 6 )
土田貴志，山本知枝，山本恵一，人見信之，小坂昇，
鹿野英士，岡田正道，小松かつ子，難波恒雄
Chemical and a n a t o m i c a l c h a r a c t e r i s t i c s o f f r e s h p e e l s o f C i t r u s s p e c i e s , i . e . C i t r u s u n s h i u , C . r e t i c u l a t a , C . s i n e n s i s , C . n a t s u d a i d a i and C . aur ‑ a n t i u m and t h e f r u i t r i p e n i n g p r o c e s s e s o f t h e s e s p e c i e s were c o m p a r e d . As r e g a r d s t h e polymethox ‑ y l a t e d f l a v o n o i d s and c o u m a r i n s , e a c h s p e c i e s had a s p e c i f i c HPLC p r o f i l e s which d i d n o t change much d u r i n g t h e m a t u r a t i o n . Each o f t h e p o l y m e t h o x y l a t ‑ e d f l a v o n o i d and coumarin c o n t e n t s reached i t s maximum d u r i n g t h e f r u i t r i p e n i n g s e a s o n , i n August O c t o b e r . The f l a v o n o i d ‑ g l y c o s i d e c o n t e n t s were t h e h i g h e s t i n young p e e l s , which d e c r e a s e d a s t h e f r u i t m a t u r e d . Anatomical v a r i a t i o n s i n t h e t h i c k n e s s o f p e e l s , t h e l e n g t h , w i d t h and s h a p e o f o i l c a v i t i e s among f r u i t s i n one t r e e , i n d i v i d u a l t r e e s i n o n e a r e a and i n d i v i d u a l a r e a s and p a r t s o f f r u i t were l e s s t h a n t h e v a r i a t i o n s i n t h e p o l y m e t h o x y l a t ‑ ed f l a v o n o i d and coumarin c o n t e n t s . E v i d e n t v a r i a ‑ t i o n s were o b s e r v e d i n t h e s e c h a r a c t e r i s t i c s and amounts o f h e s p e r i d i n c r y s t a l s a 珂 ongp e e l s from d i f f e r e n t s p e c i e s and i n d i f f e r e n t s e a s o n s . The r a t i o o f t h e s i z e o f o i l c a v i t i e s t o t h e t h i c k n e s s o f p e e l s ( l e n g t h
×w i d t h o f o i l c a v i t i e s / p e e l t h i c k n e s s ) i n ‑ c r e a s e d a s t h e f r u i t m a t u r e d . The p o l y m e t h o x y l a t e d f l a v o n o i d and coumarin c o n t e n t s gave v a l u a b l e i n f o r m a t i o n a b o u t t h e i d e n t i f i c a t i o n o f s p e c i e s and t h e a n a t o m i c a l s t r u c t u r e s a b o u t t h e maturing s t a g e .
D）和漢薬の薬効発現に関する研究
( X I ) A Purgative Action of Barbaloin i s I n ‑ duced by Eubacterium s p . Strain BAR, a Human I n t e s t i n a l Anaerobe, Capable of Transforming Barbaloin to Aloe ‑ Emodin Anthrone
B i o l . Pharm. B u l l . , 1 9 ( 1 ) , 1 3 6 〜 1 3 8 ( 1 9 9 6 ) TERUAKI AKAO, QING MING CHE, KYOICHI KOBA‑
S H I ， 恥 1ASAOHATTORI and TSUNEO N AMBA
O r a l l y a d m i n i s t e r e d b a r b a l o i n ( 1 0 0 mg/kg) d i d n o t i n d u c e any d i a r r h e a i n male Wistar r a t s , i n s p i t e o f s e v e r e d i a r r h e a w i t h s e n n o s i d e B ( 4 0 mg/kg).
A l s o , i n g n o t o b i o t e r a t s mono a s s o c i a t e d with
Pφt o s t r
ゆ， t o c o c c u s 仇 t e r m e d i u s , a human i n t e s t i n a l a n a e r o b e c a p a b l e o f r e d u c i n g s e n n i d i n s t o r h e i n a n t h r o n e , b a r b a l o i n d i d n o t i n d u c e d i a r r h e a ; t h e f a e c a l water c o n t e n t ( 7 1 . 9
%)8h a f t e r t h e a d m i n i s
司t r a t i o n o f b a r b a l o i n was n o t i n c r e a s e d , compared
w i t h t h a t ( 7 3 . 9 %) j u s t b e f o r e t h e t r e a t m e n t . How‑
e v e r , s e v e r e d i a r r h e a was i n d u c e d w i t h b a r b a l o i n i n g n o t o b i o t e r a t s mono~associated w i t h E u b a c t e r i u m s p . s t r a i n BAR, a n o t h e r human i n t e s t i n a l a n a e r o b e c a p a b l e o f t r a n s f o r m i n g b a r b a l o i n t o a l o e ‑ e m o d i n a n t h r o h e ; t h e f a e c a l w a t e r c o n t e n t was s i g n i f i c a n t l y i n c r e a s e d t o 8 5 . 5 % 8h a f t e r t h e a d m i n i s t r a t i o n , from 7 3 . 2
%b e f o r e t h e t r e a t m e n t . At t h i s t i m e , b a r b a l o i n was t r a n s f o r m e d t o a l o e ‑ e m o d i n a n t h ‑ r o n e i n t h e f e c e s from t h e g n o t o b i o t e r a t s mono a s s o c i a t e d w i t h t h e s t r a i n BAR, b u t n o t i n f e c e s from t h e c o n v e n t i o n a l r a t s o r t h e g n o t o b i o t e r a t s m o n o ‑ a s s o c i a t e d w i t h P . i n t e r m e d i u s . These f a c t s i n d i c a t e t h a t b a r b a l o i n i s i n a c t i v e a s a l a x a t i v e i t s e l f b u t i s a c t i v a t e d t o a l o e emodin a n t h r o n e , a g e n u i n e p u r g a t i v e c o m p o n e n t , by E u b a c t e r i u m s p . s t r a i n BAR.
( X I I ) Enzyme Immunoassay for Paeonimetabol ・
i n I , a Major Metabolite of Paeoniflorin by I n t e s t i n a l Bacteria
] . o f T r a d i t i o n a l M e d i c i n e s , 1 3 ( 1 ) , 7 3 〜 8 0 ( 1 9 9 6 ) MASAO HATTORI, Xru‑WEI YANG, YUE‑ZHONG SHU, OLA AHMED H E I K A L , HIROTSUGU M I Y A S H I R O , HIROMI KATO, MATAO KANAOKA, TERUAKI AKAO, KYOICHI KOBASHI and TSUNEO NAMBA
For q u a n t i t a t i v e d e t e r m i n a t i o n o f p a e o r i i m e t a ‑ b o l i n I (PM ‑
I),a m i x t u r e o f (
7R) ‑and (
75) ‑i s o m e r s t r a n s f o r m e d from p a e o n i f l o r i n ( P F ) by i n t e s t i n a l b a c t e r i a , we e s t a b l i s h e d an enzyme i m ‑ munoassay ( E I A ) method a s f o l l o w s .
As a h a p t e n , a m i x t u r e o f (
7R)‑and (
75)‑8（一2 ‑ c a r b o x y e t h y l t h i o ) p a e o n i m e t a b o l i n I (CEPM) was p r e a p a r e d by a n a e r o b i c i n c u b a t i o n o f PF with L a c t o b a c i l l u s b r e v i s i n t h e p r e s e n c e o f 3 ‑ m e r c a p t o ‑ p r o p i o n i c a c i d . Without s e p a r a t i o n o f b o t h i s o m e r s , CEPM was c o u p l e d w i t h βgalactosidase
（βGal) and b o v i n e serum a l b u m i n (BSA) l i a an N‑hydroxy‑
s u c c i n i m i d e e s t e r method t o g i v e CEPMβGal (enzyme l a b e l e d a n t i g e n ) and CEPM‑BSA ( i m ‑ munogen), r e s p e c t i v e l y . The a n t i P M I a n t i s e r u m , which had been e l i c i t e d i n r a b b i t s by i m m u n i z a t i o n w i t h CEPM‑BSA, was s p e c i f i c t o (
7R) ‑and (
75) ‑PM‑I and t h e i r 1 : 1 m i x t u r e b u t n o t t o o t h e r r e l a t e d c o m p o u n d s , when t h e a s s a y was c a r r i e d o u t by t h e d o u b l e a n t i b o d y t e c h n i q u e . S a t i s f a c t o r y s t a n d a r d c u r b e s o f ( 7 R) , (
75)PM I and b o t h i s o m e r s were
o b t a i n e d i n a r a n g e o f 0 . 1 ‑ 5 0 0 n g / t u b e b u t a p p r e ‑ c i a b l y i n t e r f e r e d by a d d i t i o n o f serum o r u r i n e t o an a s s a y m i x t u r e . By u s i n g t h e newly developed m e t h o d , q u a n t i t a t i v e d e t e r m i n a t i o n o f t h e PM
I.[ ( 7R)‑and (
75)‑ i s o m e r s ] c o n t e n t s was p o s s i b l e i n s e v e r a l f r a c t i o n s taken d u r i n g t h e i n c u b a t i o n o f PF w i t h L . b r e v i s .
E
）和漢薬の薬理作用に関する研究（日
1 )S t u d i e s on the A n t i ‑ l i p i d Peroxidative Actions of the Methanolic Extract of the Root of Aegle marmelos and I t s C o n s t i t ‑ uents In Vivo and In V i t r o
よ
o f C h i n e s e P h a r m a c e u t i c a l S c i e n c e s , 5 ( 3 ) , 1 3 2 〜 1 4 0 ( 1 9 9 6 )
Xru WEI YANG, MASAO HATTORI and TSUNEO NAMBA
The i n h i b i t o r y e f f e c t o f t h e m e t h a n o l i c e x t r a c t o f t h e r o o t o f A e g l e marmelos (MERA) and i t s c o n s t i t ‑ u e n t s on t h e l i p i d p e r o x i d a t i o n i n v i v o and i n v i t r o were s t u d i e d . The r e s u l t s s u g g e s t e d t h a t MERA i n c r e a s e d t h e a c t i v i t i e s o f s u p e r o x i d e d i s m u t a s e
(SOD) and GSH p e r o x i d a s e i n t h e l i v e r c y t o s o l o f m i c e , b u t showed no s i g n i f i c a n t e f f e c t on t h e a c t i v ‑ i t y o f c a t a l a s e , and one o f i t s major c o n s t i t u e n t s , 4
methoxy 1 m e t h y l ‑ 2 q u i n o l o n e (MMQ) i n c r e a s e d t h e a c t i v i t y o f SOD i n l i v e r t i s s u e o f mice i n t o x i c a t ed w i t h F e C l 2 a s c o r b i c a c i d (AA) ADP i n v i v o .
V a r i o u s c o n s t i t u e n t s i s o l a t e d from t h e r o o t o f t i t l e p l a n t i n h i b i t e d t h e l i p i d p e r o x i d a t i o n i n r a t l i v e r h o m o g e n a t e , which was i n v i t r o i n d u c e d by F e C l 2 ‑a s c o r b i c a c i d , C C l . ‑NADPH, o r ADP NADPH. Of t h e t e s t compounds, MMQ and i t s d e r i v a t i v e s i n t e g r i q u i n o l o n e were s i m i l a r t o αtaco‑
p h e r o l i n i n h i b i t i n g MDA p r o d u c t i o n i n r a t l i v e r microsomes induced by F e 2 + ‑a s c o r b a t e , C C 1 4 NADPH, o r ADP‑NADPH.
( XN) Suppression of H e p a t i t i s B Virus Surface Antigen Secretion by Traditional Plant Medicines
P h y t o t h e r . α ρ y R e s e a r c h , 1 0 ( 6 ) , 5 0 4 〜 5 0 7 ( 1 9 9 6 ) W AKANA GOTO, INES TOMOCO KUSUMOTO, S H I G E T O S H I KADOT A , TSUNEO N AMBA, MASAHIKO KUROKA w A and KIMIY ANU S H I R A K I
F o r t y t h r e e e x t r a c t s o f h e r b a l m e d i c i n e s were
t e s t e d f o r s u p p r e s s i n g t h e s e c r e t i o n o f h e p a t i t i s B
v i r u s (HBV) s u r f a c e a n t i g e n (HBsAg) i n v i t r o .
F o u r t e e n e x t r a c t s were f o u n d t o have a p p r e c i a b l e e f f e c t s i n t h e e x p e r i m e n t a l system u s i n g t h e PLC/
PRF / 5 c e l l l i n e . Of t h e m , t h e e x t r a c t s o f Rheum ρ
α lmatum ( r h i z o m e ) , Po
；／忽onumc u s p i d a t u m ( r o o t ) , Pan
似j a p o n i c u m ( r o o t ) and T e r m i n a l i a a r j u n a ( b a r k ) s t r o n g l y s u p p r e s s e d t h e s e c r e t i o n o f HBsAg.
The a c t i v e c o n s t i t u e n t s o f t h e s e e x t r a c t s a b s o r b e d t h r o u g h t h e g a s t r o i n t e s t i n a l t r a c t o f g u i n e a p i g s s u p p r e s s e d t h e s e c r e t i o n o f HBsAg i n t h e PLC/PRF / 5 c e l l c u l t u r e .
(XV) Ameliorating E f f e c t s of Dan‑Shen Metha‑
nol Extract on Cognitive D e f i c i e n c i e s i n Senescence‑Accelerated Mouse
K o r . ] . G e r o n t o l , 6 ( 2 ) , 1 4 〜 2 1 ( 1 9 9 6 )
TAKASHI ARIMA, lTSUKO BABA, HITOMI H O R I , YOSHIHISA KITAMURA, TSUMEO NAMBA,, MASAO HATTORI, SHIGETOSHI KADOT A and YASUYUKI NOMURA
E f f e c t o f t h e l o n g ‑ t e r m t r e a t m e n t w i t h a Dan Shen ( S a l v i a e m i l t i o r r h i z a e Radix) m
伝thanol e x t r a c t (DME) on memory and l e a r n i n g i n s c e n e s ‑ c e n c e ‑a c c e l e r a t e d mouse was i n v e s t i g a t e d by means o f Morriss water maze t a s k . DME t r e a t ‑ ment s i g n i f i c a n t l y d e c r e a s e d t h e e s c a p e l a t e n c y and f l o a t i n g t i m e i n t h e PS s t r a i n o f s e n e s c e n c e a c c e l e r ‑ a t e d mouse (SAMP8), which s t r a i n s p o n t a n e o u s l y d e v e l o p s l e a r n i n g d e f i c i t s . These r e s u l t s s u g g e s t t h a t DME t r e a t m e n t improved s p a t i a l l e a r n i n g and a c t i v a t e d e m o t i o n a l f u n c t i o n . I n neurochemical i n v e s t i g a t i o n , a l t h o u g h no e f f e c t on c h o l i n e a c e t y l t r a n s f e r a s e a c t i v i t y o r [ 3 H ] QNB ( q u i n u c l i d i n y l b e n z i l a t e ) b i n d i n g was o b s e r v e d , DME t r e a t m e n t c a u s e d i n c r e a s e s i n [ 3 H ] MK‑801 (
（十）5 ‑ m e t h y l 1 0 , 1 1 d i h y d r o 5H d i b e n z o [ a , b ] c y c l o h e p t e n ‑ 5 , 1 0 ‑ i m i n e m a l e a t e ) b i n d i n g t o t h e c o r t i c a l membrane f r a c t i o n and [ 3 H ] NNA ( N G ‑n i t r o ‑L ‑ a r g i n i n e ) b i n d i n g t o t h e c y t o s o l i c f r a c t i o n o f t h e b r a i n s t e m , and d e c r e a s e d [ 3 H ] PDBu ( p h o r b o l 1 2 , 1 3 d i b u t y r a t e ) b i n d i n g t o membranes from t h e h i p p o c a m p u s , s t r i a t u m , and c e r e b a l l u m . T h u s , o u r d a t a s u g g e s t s t h a t l o n g ‑ t e r m DME t r e a t m e n t i m p r o v e s s p a t i a l l e a r n i n g and e m o t i o n a l f u n c t i o n s i n SAMP8. As t o t h e mechanism, DME t r e a t m e n t e n h a n c e s c o r t i c a l g l u t a m a t e r g i c e x c i t a t o r y n e u r o t r a n s m i s s i o n and n i t r i c o x i d e s y n t h a s e r e l a t e d f u n c t i o n s i n b r a i n s t e m , and a f f e c t s hippocampal and s t r i a t a l f u n c ‑
t i o n s mediated by p r o t e i n k i n a s e C . F
）生薬の修治に関する研究(XVI)
Processing of Nux Vomica. V I I . Antinoci‑
c e p t i v e Effects of Crude Alkaloids from the Processed and
Unproxedded Seeds of S t r y c h n o s nux‑vomica i n Mice
B i o l . P h a r m . B u l l . , 1 9 ( 1 ) , 1 2 7 〜 1 3 1 ( 1 9 9 6 ) BAO‑CHANG C A I , TETSURO NAGASAWA, SHIGETO‑
S H I KADOT A , MASAO HATTORI, TSUNEO N AMBA and Y A S U S H I KURAISHI
We examined t h e a n t i n o c i c e p t i v e e f f e c t s o f t h e c r u d e a l k a l o i d f r a c t i o n s (CAF) o f nux vomica ( t h e d r i e d s e e d s o f S t r y c h n o s n
似v o m i c a
L.)and t h e i n f l u e n c e d o f v a r i o u s p r o c e s s i n g methods upon t h e i r a n t i n o c i c e p t i o n i n t h r e e a n a l g e s i c t e s t s i n m i c e . I n t h e t a i l p r e s s u r e t e s t , t h e CAF ( 0 . 0 1 ‑ 1 μg/kg, i . p . ) o f nux vomica t h a t was u n p r o c e s s e d o r t r e a t e d w i t h s a n d , l i c o r i c e , o i l o r v i n e g a r and sand p r o c e s s i n g showed c l e a r a n t i n o c i c e p t i o n . The CAF ( 1 μg/kg, i . p . ) o f v i n e g a r p r o c e s s e d nux vomica showed a n t i n o c i c e p t i o n , w i t h o u t e f f e c t s a t l o w e r d o s e s o f 0 . 0 1 and 0 . 1 μg/kg and t h o s e t r e a t e d w i t h u r i n e ‑o r u r i n e and sand p r o c e s s i n g were w i t h o u t e f f e c t s a t d o s e s o f 0 . 0 1 1 μg/kg. Morphine ( 2 mg/kg, s . c . ) showed s h o r t ‑l a s t i n g a n t i n o c i c e p t i o n , without e f f e c t s a t a d o s e o f 1 μ g / k g . I n t h e h o t p l a t e t e s t , t h e CAF ( 1 0 0 μ g / k g , i . p . ) o f nux vomica h a v i n g u n d e r g o n e sand p r o c e s s i n g produced a s i g n i f i c a n t a n t i n o c i c e p t i o n , w i t h o u t e f f e c t s a t l o w e r d o s e s o f 0 . 0 1 and 1 μg/kg. The CAF ( 0 . 0 1 ‑ 1 0 0 μ g / k g , i . p . ) o f nux vomica t h a t was u n p r o c e s s e d o r t r a t e d w i t h o i l o r v i n e g a r and sand p r o c e s s i n g and morphine ( 1 and 1 0 0 μg/kg, s . c . ) were w i t h o u t e f f e c t s . I n t h e a c e t i c a c i d i n d u c e d w r i t h i n g t e s t , t h e CAF ( 1 μg/
k g , i . p . ) o f nux vomica t h a t was t r e a t e d w i t h sand
p r o c e s s i n g s i g n i f i c a n t l y i n h i b i t e d t h e writhing
b e h a v i o r , w h i l e t h o s e o f nux vomica t h a t was u n ‑
p r o c e s s e d o r t r e a t e d w i t h o i l o r v i n e g a r and sand
p r o c e s s i n g and morphine were w i t h o u t e f f e c t s a t a
d o s e o f 1 μg/kg. The p r e s e n t r e s u l t s d e m o n s t r a t e
t h e a n t i n o c i c e p t i v e e f f e c t s o f t h e CAF o f nux
vomica and s u g g e s t t h a t s a n d ‑ p r o c e s s i n g i s good
f o r t h e a n a l g e s i c p o t e n c y o f nux v o m i c a . I
ti s a l s o
s u g g e s t e d t h a t t h e CAF o f nux vomica h a s d i s t i n c t
a n t i n o c i c e p t i v e p o t e n c y , e v e n a f t e r t r e a t m e n t w i t h
l i c o r i c e
，一o i l ‑ ,v i n e g a r and sand p r o c e s s i n g .
G) HIV
プロテアーゼ阻害活性を有する生薬 の探索( X V I I ) Screening of Egyptian Folk Medicinal Plant Extracts for Anti‑human Im‑
munodeficiency Virus Type‑1 (HIV ‑1) Activity
] . o f T r a d i t i o n a l M e d i c i n e s , 1 3 ( 1 ) , 5 9 〜 6 5 ( 1 9 9 6 ) TAKUYA KAWAHATA, TORU OTAKE, HARUYO M O R I , MOTOKO MORIMOTO, NOBORU UEBA, INES TOMOCO KUSUMOTO, SAHAR EL MEKKAWY, MASAO HATTORI and TSUNEO N AMBA
As a r e s u l t o f s c r e e n i n g t h e e x t r a c t s o f 4 1 E g y p t i a n f o l k m e d i c i n e s , s i x p l a n t s showed i n h i b i ‑ t o r y a c t i v i t y a g a i n s t HIV‑1 : methanol e x t r a c t s o f A b r u s
ρr e c a t o r i u s
L.,A r t e m i s i a a b s i n t h i u m
L.,C r o t o n t i g l i u m
L.and D a t u r a s t r a m o n i u m
L.,and w a t e r e x t r a c t s o f B a s s i a m u r i c a t a
(L.)MURR., C e n t a u r e a s c o ρ a r i a
L.,C r o t o n t i g l i u m
L. andD a t u r a s t r a m o n i u m
L.,T h e i r : ‑ e s p e c t i v e a n t i HIV 1 a c t i v ‑ i t y ( I C 5 0 ) f o r MT‑4 c e l l s were 2 . 0 , 9 . 8 , 0 . 0 2 5 , 4 . 1 , 1 3 5 , 4 6 . 2 , 2 . 0 and 9 0 . 0 ( μ g / m l ) . C r o t o n t i g l i u m
L.a l s o s u p p r e s s e d g i a n t c e l l f o r m a t i o n i n c o c u l t u r e s o f MOLT 4 c e l l s w i t h MOLT‑4/HTLV I l l s c e l l s . M o r e o v e r , Abrus ρ r e c a t o r i u s
L.,A r t e m i s i a a b s i n t h ‑ ium L . , C r o t o n t i g l i u m
L.had d i r e c t e f f e c t s and d e c r e a s e d t h e i n f e c t i v i t y o f HIV‑1. However, none o f them showed any i n h i b i t o r y e f f e c t on t h e r e v e r s e t r a n s c r i p t a s e and p r o t e a s e a c t i v i t y o f HIV 1 .
H）和漢薬の生理活性成分の研究
(XVIII)
Effects of Alkaloids from Corydalis decumbens on Contraction and Electro‑
physiology of Cardiac Myocytes P h y t o t h e r a ρy R e s e a r c h , 1 0 ( 1 ) , 1 8 〜 2 2 ( 1 9 9 6 ) SHIGETOSHI KADOTA, XIAO L I SUN, PURUSOTAM BASNET, TSUNEO NAMBA and YASUNORI MOMOSE
The c h l o r o f o r m e x t r a c t o f C o r y d a l i s d e c u m b e n s s i g n i f i c a n t l y i n c r e a s e d t h e b e a t i n g a m p l i t u d e o f c u l t u r e d m y o c a r d i a l c e l l s h e e t s . Chemical a n a l y s i s l e d t o t h e i s o l a t i o n o f i s o q u i n o l i n e and p r o t o p i n e a l k a l o i d s . Of t h e s e i s o l a t e d a l k a l o i d s , c o r l u m i d i n e and ( +) a d l u m i d i n e i n c r e a s e d t h e b e a t i n g a m p l i ‑ t u d e , b u t
（十）e g e n i n e d e c r e a s e d t h e b e a t i n g r a t e and b e a t i n g a m p l i t u d e w h i l e p r o t o p i n e d i d n o t show any a c t i v i t y . We a l s o s t u d i e d t h e e f f e c t s of 弘 （ ＋ ） e g e n i n e and c o r l u m i d i n e on c o n t r a c t i l e r e s p o n s e s and C a 2 + c u r r e n t s i n s i n g l e b u l l f r o g a t r i a l c e l l s u s i n g
t h e v o l t a g e clamp m e t h o d . (
+) ‑ E g e n i n e i n h i b i t e d C a 2
十c u r r e n tby 6 8
%o f t h e c o n t r o l i n s i n g l e c e l l o f b u l l f r o g a t r i u m , w h i l e c o r l u m i d i n e i n c r e a s e d C a 2 + c u r r e n t t o 6 0
%a t a c o n c e n t r a t i o n o f 0 . 0 3 mM.
( X I X 〕 ProtectiveEffect of C e l o s i a n , an Acidic Polysaccharide, on Chemically and Im‑
munologically Induced Liver I n j u r i e s B i o l . P h a r m . B u l l . , 1 9 ( 4 ) , 5 6 7 〜 5 7 2 ( 1 9 9 6 ) Kon HASE, SHIGETOSHI KADOT A , PURUSOT AM BASNET, TOORU TAKAHASHI and TSUNEO NAMBA
H e p a t o p r o t e c t i v e e f f e c t o f c e l o s i a n , an a c i d i c p o l y s a c c h a r i d e i s o l a t e d from t h e water e x t r a c t o f t h e s e e d o f C e l o s i a a r g e n t e a , was i n v e s t i g a t e d u s i n g c h e m i c a l and i m m u n o l o g i c a l l i v e r i n j u r y m o d e l s . C e l o s i a n i n h i b i t e d t h e e l e v a t i o n o f serum enzyme (GPT, GOT, LDH) and b i l i r u b i n l e v e l s on carbon t e t r a c h l o r i d e ( C C l 4
）一i n d u c e dl i v e r i n j u r i e s i n r a t .
Ina d d i t i o n , t h e h e p a t o p r o t e c t i v e e f f e c t o f c e l o s i a n was a l s o o b s e r v e d i n t h i s model o f l i v e r i n j u r y by h i s ‑ t o p a t h o l o g i c a l f i n d i n g s . M o r e o v e r , c e l o s i a n s u p p r e s ‑ s e d r i s e s i n GPT o r m o r t a l i t y on f u l m i n a n t h e p a t i t i s i n d u c e d by D g a l a c t o s a m i n e / l i p o p o l y s a c c h a r i d e ( D Ga!N
ILPS) o r P r o p i o n i b a c t e r i u m a c η e s
ILPS i n m i c e . These f i n d i n g s s u g g e s t e d t h a t c e l o s i a n i n an a c t i v e component i n p r o t e c t i o n a g a i n s t c h e m i c a l and i m m u n o l o g i c a l h e p a t i t i s and t h e a c t i v i t y was f o u n d t o be a d o s e d e p e n d e n t .
C e l o s i a n showed a c o n c e n t r a t i o n d e p e n d e n t i n h i b ‑ i t o r y e f f e c t on l i p i d p e r o x i d e (LPO) g e n e r a t i o n i n v i t r o . Though c e l o s i a n d i d n o t r e d u c e t h e r e l e a s e o f tumor n e c r o s i s f a c t o r α
（TNF
−α
） ,i t p r o t e c t e d a g a i n s t r e c o m b i n a n t human TNFα
（rhTNFα）
i n d u c e d l i v e r i n j u r y i n D g a l a c t o s a m i n e s e n s i t i z e d m i c e .
(XX) Potent Antihepatotoxic Activity of Dicaffeoyl Quinic Acids from Propolis B i o l . P h a r m . B u l l . , 1 9 ( 4 ) , 6 5 5 〜 6 5 7 ( 1 9 9 6 ) PURUSOTAM BASNET, KATSUMICHI 恥 1A T S U S H I G E , KOJI HASE, SHIGETOSHI KADOT A and TSUNEO NAMBA
H e p a t o p r o t e c t i v e a c t i v i t y g u i d e d c h e m i c a l a n a ‑
l y s e s l e d t o t h e i s o l a t i o n o f two d i c a f f e o y l q u i n i c
a c i d d e r i v a t i v e s , m e t h y l 3 , 4 d i 0 c a f f e o y l q u i n a t e
( 1 ) and 3 , 4 ‑ d i
0 ‑c a f f e o y
1q u i n i c a c i d ( 2 ) from
water e x t r a c t o f p r o p o l i s , and t h e i r s t r u c t u r e s were
d e t e r m i n e d by t h e u s e o f 2D NMR. These com‑
p o u n d s were s t r o n g e r a n t i h e p a t o t o x i c a g e n t s t h a n g l y c y r r h i z i n .
(XX!)
Hepatoprotective Effects of Pan α x no t o
・gineseng : Ginsenosides ‑Re and ‑Rg
1a s i t s Active Constituents i n D ‑g a l a c t o s ‑ arnine/Lipopolysaccharide‑induced Liver Injury
P h y t o m e d i c i n e , 2 ( 4 ) , 2 9 7 〜 3 0 3 ( 1 9 9 6 )
]EEVAN KUMAR P R A S A I N , S H I G E T O S H I KADOTA, PURUSOTAM BASNET, Kon HASE and TSUNEO NAMBA
The h e p a t o p r o t e c t i v e e f f e c t s o f t h e methanol and water e x t r a c t s o f t h e r o o t s o f Panax n o t o g i n s e n g were s t u d i e d on v a r i o u s a n i m a l m o d e l s . The meth
回a n o l and w a t e r e x t r a c t s o f P . n o t o g i n s e n g showed a s i g n i f i c a n t a c t i v i t y i n c a r b o n t e t r a c h l o r i d e ‑ i n d u c e d l i v e r i n j u r y i n r a t s . However, t h e methanol e x t r a c t was f o u n d t o b e more a c t i v e . The h e p a t o p r o t e c t i v e e f f e c t s o f t h e methanol e x t r a c t was f u r t h e r e v a l u a t ‑ e d u s i n g D g a l a c t o s a m i n e a s w e l l a s h e a t k i l l e d Pro ρ i o n i b a c t e r i u m a c n e s
Il i p o p o l y s a c c h a r i d e i n ‑ d u c e d l i v e r i n j u r y m o d e l s . Two major i s o l a t e s from t h e m e t h a n o l e x t r a c t , g i n s e n o s i d e s Re and R g , , showed a s i g n i f i c a n t h e p a t o p r o t e c t i v e e f f e c t on D ‑ g a l a c t o s a m i n e
Il i p o p o l y s a c c h a r i d e i n d u c e d l i v e r mJury m m i c e .
( X X I I ) Hepatoprotective Effects of Traditional M e d i c i n e s . I s o l a t i o n of the Active Con‑
s t i t u e n t s from Seeds of C e l o s i , αargentea P h y t o t h e r a
ρyR e s e a r c h , 1 0 ( 5 ) , 3 8 7
〜39 2 ( 1 9 9 6 ) K O J I HASE, S I J G E T O S H I KODOTA, PURUSOTAM BASMET, TOORU TAKAHASHI and TSUNEO NAMBA
The h e p a t o p r o t e c t i v e e f f e c t s o f water e x t r a c t s from t w e l v e c r u d e d r u g s were i n v e s t i g a t e d . Among t h e m , t h e water e x t r a c t s o f C a s s i a o b t u s i f o l i a , C e l o s i a a r g e n t e a , C u c u r b i t a m o s c h a t a and Curcuma a e r u g i n o s a showed a s i g n i f i c a n t p r o t e c t i v e e f f e c t on c a r b o n t e t r a c h l o r i d e i n d u c e d l i v e r i n j u r y i n r a t s and D g a l a c t o s a m i n e ( D GaIN)
Il i p o p o l y s a c c h a r i d e ( L P S ) i n d u c e d l i v e r i n j u r y i n m i c e . The water e x t r a c t o f
C. ar g e n t e a was f o u n d t o b e t h e most e f f e c t i v e . The h e p a t o p r o t e c t i v e a c t i v i t y g u i d e d f r a c t i o n a t i o n o f t h i s e x t r a c t l e d t o t h e i s o l a t i o n o f an a c t i v e c o n s t i t u e n t . T h i s component was named a s c e l o s i a n , and was f o u n d t o b e an a c i d i c h e t e r o ‑ g l y c a n ( m o l e c u l a r w e i g h t ・ 1 . 9
×1 0 5 ) c o n t a i n i n g 4
%o f p r o t e i n s .
( XXITI) Four Di‑O‑caffeoyl Quinic Acid Deriva‑
