Evaluation of glycyrrhizin variation in Gancao collected from 1986 to 2000 at drug markets in Japan and discrimination of Daitou-Gancao and Dongbei-Gancao

Quality variation of 337 lots of medicinal Gancao (Kanzo in Japanese) collected at drug markets in Japan from 1986 to 2000 was evaluated by measuring glycyrrhizin (GL)-content and roots specific gravity (RSG). The present results clearly indicated that GL-content (5.16±1.00%, 109 lots) of Dongbei-Gancao (Tohoku-Kanzo in Japanese) is significantly higher (4.36±1.45%, 228 lots) than that of Xibei-Gancao (Seihoku-Kanzo in Japanese). Furthermore, the RSG of Dongbei-Gancao (0.51±0.09 g / cm^3, 66 roots) is significantly lower than that of Xibei-Gancao (0.65±0.11 g / cm^3, 66 roots). This finding indicates that the RSG is useful indicator in distinguishing Dongbei-Gancao with flexible roots from Xibei-Gancao with hard and heavy roots. During the experimental process, it was found that there have been some odd Dongbei-Gancao with high RSG in drug markets in Japan from about 1976. Our market research in China indicated that Gancao with knot-like upper roots collected at Xibei (northwest) region in China has been under circulation designated as Daitou (or Youtou)-Gancao from early 1990\u27s in China. The result of RSG examination on Daitou-Gancao (Taito- or Yuto-Kanzo in Japanese) allows us to consider that some Dongbei-Gancao with high RSG seems to correspond to Daitou-Gancao. This is the first report to characterize Daitou-Gancao which has a similar knot-like upper root and high GL-content to Dongbei-Gancao and also has RSG as high as Xibei-Gancao. 本論文の特徴は,日本市場甘草の品質変動を1986年から2000年まで15年間にわたって集めた多数の試料(337 lots)に基づいて検討した点にある。比較のために同定可能な花期・果実期に中国内蒙古自治区の東部(翁牛特旗頭道溝:東北甘草の産地)で採集しGlycyrrhiza uralensis FISCH.の根を用いた。市場品甘草のglycyrrhizin含量は年度によって変動するが,15年間の平均値において東北甘草のglycyrrhizin含量は西北甘草より有意に高いことを確認した。見掛け比重(RSG)は根を円柱とみなして重量と体積を量り算出した。東北甘草のRSGは西北甘草より有意に低いことを確認した。今回確立したRSGは,柔軟な東北甘草と質の充実した堅い西北甘草を判別する経験的な選品法を裏付ける指標となる。なお1997年頃以降に集めた東北甘草の中にRSGの高い(mean+2S.D.を超える)個体が散見された。中国における市場調査の結果,甘粛省など中国西北地方で産し根頭部に「こぶ」のある甘草が帯頭甘草あるいは有頭甘草という名で出回り,日本に輸出されていることが明らかになった。市場品の東北甘草中に散見されたRSGの高い甘草は,帯頭甘草の可能性があることをShapiro-Wilk検定で統計学的に推定した。この帯頭甘草(甘粛省産)を入手して分析したところ,根頭部に「こぶ」のある形態とglycyrrhizin含量の高い点は東北甘草に類似し,RSGが高い点は西北甘草に類似することが明らかになった。帯頭(有頭)甘草(中国市場名)の流通状況や形質を明らかにしたのは本論文が最初である

Growth and glycyrrhizin contents in Glycyrrhiza uralensis roots cultivated for four years in eastern Nei-Meng-gu of China

In China, the collection of wild Glycyrrhiza roots came to be restricted since there is a high possibility of inducing desertification. In order to compensate for the insufficiency of wild Glycyrrhiza resources with cultivated ones, the cultivation study of Glycyrrhiza uralensis has been carried out since 1998 and is still continuing. The cultivation field is located in the eastern region of Nei-Meng-gu, inner Mongolia, which is the habitat of G. uralensis and the major source of Dongbei-Gancao, Tohoku-Kanzo in Japanese. The seeds of G. uralensis were sown on May in 1998 and seedling roots were transplanted to the field the next May. The glycyrrhizin (GL)-content of 4-year-old lateral roots, which were newly grown from the transplanted seedling taproot, exceeded the Japanese Pharmacopoeia XIV standard (2.5% or more GL). This is the first report to examine GL-content of G. uralensis cultivated in northeast China related to cultivation years. However, since GL-content (2.81 ± 0.76%) of 4-year-old lateral roots was at a low level yet, further examinations on improvement of cultivation conditions are in progress to obtain suitable length and thickness of lateral roots, high GL-contents and adequate ingredients-composition similar to those of wild Glycyrrhiza roots currently in use. 中国において野生甘草(Gancao)の採集が砂漠化を誘発するために2000年に制限する通知が出され,栽培甘草の実用化が強く望まれるようになっている。本研究は,地球環境を保全し甘草資源を確保(野生甘草の不足分を栽培甘草で補填)する観点から企画された。本論文の特徴は,Glycyrrhiza uralensis Fisch.を内蒙古自治区の東部(元宝山区古山鎮西六家村:東北甘草の産地)において栽培した点にある。なお試験植物がG.uralensisであることは,3-4年目の夏に開花した花と果実および小葉の形態から確認した。5月に播種し翌年の5月に苗を堀り上げ根を20-25cmに切りそろえた後,移植した。移植根から新たに生育した4年目の側根(lateralroots)のglycyrrhizin(GL)含量が,日本薬局方の基準値(JPXIV:GLを2.5%以上含む)を越えることが明らかになった。中国の東北甘草の生産地域で栽培されたG.uralensisのGL含量を栽培年数と関連付けて検討したのは,本論文が初めてである。今回の実験によって,栽培甘草が薬用資源として利用できる可能性が高まった。しかしながら,4年目のGL含量(2.81±0.76%)は日本市場の東北甘草(Dongbei-Gancao)の平均値(5.16±1.00%)と比べて満足できる値に達していない。優良種苗の選抜や栽培方法の改良など今後の検討が必要である。この栽培実験と併行して中国の甘草生産地において栽培されたGlycyrrhiza根を調査したところ,JPXIV基準に適合する物は得られなかった。なお,西北地域で蒐集した試料は移植せずに5-6年間栽培された主根(tap-roots)であった。今回我々が実施した栽培実験においてJPXIVの基準に適合したのは,移植後に主根から生育した側根(lateralroots)であった。移植の有無や移植時期が側根の成長やGL含量に及ぼす影響に関しては,現在も内蒙古東部の畑において検討中である

Organ selectivity of Juzen-taiho-to and Ninjin-yoei-to in the expression of anti-metastatic efficacy

本研究では,マウス結腸癌Colon26-L5細胞の門脈内移入により形成される肝転移および同細胞の尾静脈内移入により形成される肺転移に対する,十全大補湯および人参養栄湯の経口投与による抑制効果を検討した。十全大補湯は肝転移に対して有意に抑制したが,肺転移には抑制効果を示さなかった。これに対して,人参養栄湯は逆の効果,すなわち肝転移には効果を示さなかったが,肺転移に対して有意な抑制を示した。このように,同一の癌細胞と同系のマウスの転移病態モデルを用いて,二つの方剤の臓器選択的な転移抑制効果が観察された。このような効果発現の差異の解釈は,13世紀に確立されたtheory of Jingand Lun(引経報使)の考えに,部分的に通ずるものがあると思われる。 We investigated the inhibitory effect of oral administration of Juzen-taiho-to and Ninjin-yoei-to on liver metastasis caused by intraportal vein injection of colon 26-L5 cells and lung metastasis by intravenous injection of same tumor cells. Juzen-taiho-to significantly inhibited liver metastasis but not lung metastasis. In contrast, Ninjin-yoei-to was effective at inhibiting lung metastasis but not liver metastasis. In the experimental liver and lung metastases model using same tumor in syngeneic mice system, oral administration of both formulations showed a differential pattern with organ selectivity for the expression of anti-metastatic effects. These results suggest that the different expression of the anti-metastatic effects of both Kampo medicines on tumor metastasis are partly based on the medicinal guides according to the theory of Jing and Lun (Inkei-hoshi) formed in the 13th century

Accidental plant poisoning and identification of Datura seed

Recently an accidental plant poisoning occurred in Niigata. In order to identify the actual cause of poisoning, we analyzed the remaining sample, which caused the poisoning. Based on the patient\u27s symptoms such as a dilatation of the pupil, thirst, and others, the poisoning was recognized as being caused by an anticholinergic substance. Furthermore, compressed-reniform shape and the buff color of the sample suggested the origin to be derived from the seeds of Datura (Solanaceae) . The studies were carried out by comparing the external features and anatomical characteristics of the seeds with four comparative materials, namely D. innoxia, D. metel, D. stramonium, and D. stramonium f. inermis. These four materials could be distinguished from each other externally based on the differences in the color and texture, and anatomically by the structure of the sclerenchymatous epidermal cells. In conclusion, the poisoning was confirmed to be caused by an accidental intake of the seeds of D. innoxia. 新潟で中毒事故が発生した。この中毒の真の原因を明らかにするため,我々は患者が摂取した残りの食物試料を分析した。患者の諸症状,即ち,瞳孔散大,口渇等から,この中毒は抗コリン性物質により引き起こされたものと考えられた。また,患者が摂取した残りの試料は,扁平かつ腎臓形で黄土色を呈していたことから,ナス科 Datura 属の種子が原因と考えられた。そこで,比較材料4種即ち,D.inoxia ,D.metal ,D.sramonium f.inermis とともに,組織形態を含む比較形態学的研究を行った。これら4分類群(taxa)の種子は外部形態的には色と質感,内部形態的には表皮の構造の違いによって区別しえた。結論として,この中毒はD.inoxia の種子の誤摂取によって引き起こされたものと確証した

Effect of some Kampo medicines, including Tokaku-joki-to (Tao-He-Cheng-Qi-Tang), on IgE-mediated triphasic skin reaction in passively sensitized mice

Previous studies have reported that the mice passively sensitized with anti-DNP IgE antibody exhibited IgE-mediated biphasic cutaneous reaction with an immediate phase response (IPR) at 1 h and a late phase response (LPR) at 24 h after the challenge of DNFB (dinitrofluorobenzene) . We recently found that the third phase inflammatory response with intense and persisting infiltration of eosinophils, named very late phase response (vLPR) , was induced following IPR and LPR in response to DNFB in passively sensitized mice, and that the peak response of vLPR was on the 8^ day after the challenge. The inhibitory effect of Kampo medicines on the triphasic cutaneous inflammatory reaction was divided into several groups in terms of their inhibition rate of ear swelling. Among the formulations, Tokaku-joki-to (Tao-He-Cheng-Qi-Tang) was effective at inhibiting IPR, LPR and vLPR ( +/+/+ group) and scratching behavior in IPR. The Inhibitory effect of Tokaku-joki-to on triphasic cutaneous reaction primarily depends on its composed crude drugs, Glycyrrhizae Radix and Cinnamomi Cortex. These findings indicate that Tokaku-joki-to formulation is usefull for the inhibition of cutaneous inflammatory diseases. 抗DNP IgE抗体で受動感作したマウスの耳介にDNFB(ジニトロフルオロベンゼン)を塗布することにより,1時問およぴ24時間目をピークとする即時相反応(IPR)およぴ遅発相反応(LPR)からなるIgE介在二相性皮膚反応を示すことがすでに知られている。我々は最近,この受動感作マウスにおいてDNFBによる反応惹起後にIPR,LPRに続く,三相目の強い炎症性反応を見出し,超遅発相反応(vLPR)と名付けた。これは抗原塗布から8日目をピークとする,著明かつ持続的な好酸球の浸潤を伴う腫脹反応である。種々の漢方方剤を用いて,この三相性皮膚反応に対する抑制効果を検討した結果,各相の耳介腫脹の抑制率に基づき,いくつかのグループに分類された。検討した方剤中,桃核承気湯はIPR,LPR,vLPRの三相反応に対して抑制を示し(+/+/+群),さらにIPRで観察される耳介の掻き行動(痒みの指標と考えられる)を抑制した。桃核承気湯の三相性皮膚反応に対する抑制効果の発現は,主として構成生薬である甘草およぴ桂皮に基づくことが示唆された。これらの知見から,漢方方剤:桃核承気湯が炎症性皮膚疾患に有効であることが示された

Field study and pharmaceutical evaluation of Glycyrrhiza uralensis roots cultivated in China(Material study)

Medicinal licorice (underground parts of Glycyrrhiza plants) is an important crude drug prescribed in many traditional Chinese formulations. Recently in China, the collection of wild Glycyrrhiza plants was restricted in 1984 because it promotes desertification. With the shortage of licorice due to this restriction, there has been a demand for the cultivation of Glycyrrhiza plants. In the former half of this review, we summarized the results of our survey of the cultivation/production state of G. uralensis in China that was performed from 1998 to 2001. In China, the cultivation of G. uralensis was initiated in the early 1990s. However, the glycyrrhizin (GL) content of the cultivated roots (mainly 3-year-old roots) we collected in our survey in China did not fulfill the standard (≥2.5%) of the Japanese Pharmacopoeia (JP XIV). Such roots cannot be used for medicine in Japan. In the latter half of this review, the results of our experiments of G. uralensis cultivation in eastern Neimenggu autonomy in China were summarized. We performed cultivation from seeds of G. uralensis, and taproots were transplanted the next year. As a result, adventitious roots collected in autumn in the 4th year after seeding conformed to the JP XIV standard for GL content. This study was the first to clarify the appropriateness of G. uralensis roots cultivated in China as a resource of medicinal licorice. The anti-allergic action of the cultivated roots and their GL bioavailability after oral administration were similar to those of existing medicinal licorice prepared from wild G. uralensis in China. These results suggest that cultivated G. uralensis roots can compensate for the insufficient licorice resources. This study not only aims at the development of licorice resources and the assessment of their standards but is also a phyto-remediation study on the global environment using cultivated plants