Lv Chao-Geng, Wang Sheng, Yang Wan-Zhen, Liu Tan, Guo Lan-Ping
National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, State Key Laboratory Breeding Base of Dao-di Herbs, Beijing 100700, China.
School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China.
Zhongguo Zhong Yao Za Zhi. 2017 Apr;42(8):1566-1571. doi: 10.19540/j.cnki.cjcmm.20170222.015.
The accumulation of rosmarinic acid, acetylshikonin, deoxyshikonin, β, β'-dimethylacrylshikonin and isovalerylshikonin was investigated in cell suspension cultures of Arnebia euchroma (Royle) Johnst under the influence of 2-aminoindan-2-phosphonic acid (AIP), an inhibitor of phenylalanine ammonia lyase (PAL), and of the effector methyl jasmonate (MeJA). The results showed that methyl jasmonate promoted the accumulation of rosmarinic acid and shikonin derivatives. Conversely, 2-aminoindan-2-phosphonic acid suppressed the formation of rosmarinic acid, which indicated that AIP, indeed, was able to inhibit the phenylpropanoid pathway in A. euchroma. Meanwhile, the content of total shinkonins and other four kinds of shikonin derivatives, though varied in degrees, was also inhibited. And the inhibition was dose-dependent and time-dependent. Acetylshikonin responsed most rapidly to the treatment of AIP, the content reduced after 24 h of treatment and decreased to only half of those untreated control 48 h after teratment. β, β'-Dimethylacrylshikonin, difffer from acetylshikonin, responded much slowly to the treatment, inhibition could only be observed 96 h later. These suggest that phenylpropanoid pathway plays an important role in the shikoninsbiosynthesis, and this study provides a reference for the further research in metabolic regulation of producing shikonins by cell culture technology and biosynthesis pathways of shikonin derivatives. Still, shikonins biosynthesis pathways is complicated, the exact dose- and time-effect relationship of AIP and interaction between AIP and other effectors like MeJA need further research.
研究了在苯丙氨酸解氨酶(PAL)抑制剂2-氨基茚-2-膦酸(AIP)和效应物茉莉酸甲酯(MeJA)的影响下,新疆紫草(Arnebia euchroma (Royle) Johnst)细胞悬浮培养物中迷迭香酸、乙酰紫草素、脱氧紫草素、β,β'-二甲基丙烯酰紫草素和异戊酰紫草素的积累情况。结果表明,茉莉酸甲酯促进了迷迭香酸和紫草素衍生物的积累。相反,2-氨基茚-2-膦酸抑制了迷迭香酸的形成,这表明AIP确实能够抑制新疆紫草中的苯丙烷类途径。同时,总紫草素和其他四种紫草素衍生物的含量虽然有不同程度的变化,但也受到了抑制。并且这种抑制具有剂量依赖性和时间依赖性。乙酰紫草素对AIP处理的反应最为迅速,处理24小时后含量降低,处理48小时后降至未处理对照的一半。β,β'-二甲基丙烯酰紫草素与乙酰紫草素不同,对处理的反应要慢得多,96小时后才观察到抑制作用。这些表明苯丙烷类途径在紫草素生物合成中起重要作用,本研究为进一步研究细胞培养技术生产紫草素的代谢调控和紫草素衍生物的生物合成途径提供了参考。不过,紫草素生物合成途径复杂,AIP的确切剂量和时间效应关系以及AIP与MeJA等其他效应物之间的相互作用还需要进一步研究。