Co-cultured adventitious roots of Echinacea pallida and Echinacea purpurea inhibit lipopolysaccharide-induced inflammation via MAPK pathway in mouse peritoneal macrophages / 中草药·英文版

Objective: In order to elucidate the biological activity of the co-cultured adventitious roots (ARs) of Echinacea pallida and Echinacea purpurea and provide theoretical basis for its application, and the anti-inflammatory activities and potential mechanisms of co-cultured ARs were studied. Methods: The experimental materials were obtained by bioreactor co-culture technology and used in the activity research. In this study, mouse macrophages induced by lipopolysaccharide (LPS) were used as in vitro model. Different concentrations of AR extract (50–400 g/mL) were used to treat cells. The expression of pro-inflammatory cytokines was determined using enzyme linked immunosorbent assay. The inducible nitric oxide synthase and cyclooxygenase-2 expression, mitogen-activated protein kinase (MAPK) phosphorylation, and the inhibitor of nuclear factor-kappa B-α levels were determined by the Western blot analysis. Results: In the co-cultured ARs, total flavonoids and total caffeic acid were determined, and the contents of both bioactive compounds were significantly higher than those ARs from the single-species culture. Compared with the control group, the large amount of pro-inflammatory mediators was released after LPS stimulation. However, in the extract groups with different concentrations (25, 50, and 100 g/mL), the production of these pro-inflammatory mediators was inhibited in a dose-dependent manner. Furthermore, the levels of phosphorylation of MAPK proteins, including p-p38, p-c-Jun N-terminal kinase, and p-extracellular regulated protein kinases were significantly (P < 0.05) decreased in the extract groups, revealing that the AR extract probably involved in regulating the MAPK signaling pathway. Conclusion: Collectively, our findings suggested that the co-cultured ARs of E. pallida and E. purpurea can inhibit production of pro-inflammatory mediators in mouse peritoneal macrophages and possess the anti-inflammatory effect by regulating MAPK signaling pathways.

Optimization of Extraction Condition of Methyl Jasmonate-treated Wild Ginseng Adventitious Root Cultures using Response Surface Methodology

The usage of wild ginseng (Panax ginseng C.A. Meyer) has been limited due to short supply and high price. Therefore, sufficient production as well as efficient extraction of mountain ginseng are required for the development as products. In this study, wild ginseng adventitious root cultures were prepared for efficient production with advantages of fast growth and stable production. Treatment of methyl jasmonate (MJ) to wild ginseng adventitious root cultures increased the extraction yield and antioxidative activity. Further investigation on effect of extraction conditions suggested the importance of ethanol concentration on antioxidative activity and extraction yield of MJ-treated wild ginseng adventitious root cultures. Optimized extraction condition of MJ-treated wild ginseng adventitious root cultures for maximum extraction yield and antioxidative activity was determined using response surface methodology with three-level-three-factor Box-Behnken design (BBD). Extraction of 1 g MJ-treated wild ginseng adventitious root culture with 30 ml of 9% ethanol at 30 ℃ produced 310.2 mg extract with 71.0% antioxidative activity at 100 µg/ml. Taken together, MJ-treated wild ginseng adventitious root culture is valuable source for wild ginseng usage and optimized extraction condition can be used for the development of functional products or folk remedies.

Morpho-anatomical study of Stevia rebaudiana roots grown in vitro and in vivo

ABSTRACT Stevia rebaudiana (Bertoni) Bertoni, Asteraceae, is used as a food additive because its leaves are a source of steviol glycosides. There are examples of tissue culture based on micropropagation and phytochemical production of S. rebaudiana leaves but there are few studies on adventitious root culture of S. rebaudiana. More than 90% of the plants used in industry are harvested indiscriminately. In order to overcome this situation, the development of methodologies that employ biotechnology, such as root culture, provides suitable alternatives for the sustainable use of plants. The aim of this study was to compare morpho-anatomical transverse sections of S. rebaudiana roots grown in vitro and in vivo. The in vitro system used to maintain root cultures consisted of a gyratory shaker under dark and light conditions and a roller bottle system. Transverse sections of S. rebaudiana roots grown in vitro were structurally and morphologically different when compared to the control plant; roots artificially maintained in culture media can have their development affected by the degree of media aeration, sugar concentration, and light. GC–MS and TLC confirmed that S. rebaudiana roots grown in vitro have the ability to produce metabolites, which can be similar to those produced by wild plants.

Blood flow-improving activity of methyl jasmonate-treated adventitious roots of mountain ginseng / 한국실험동물학회지

Ginsenosides from Panax ginseng are well known for their diverse pharmacological effects including antithrombotic activity. Since adventitious roots of mountain ginseng (ARMG) also contain various ginsenosides, blood flow-improving effects of the dried powder and extract of ARMG were investigated. Rats were orally administered with dried powder (PARMG) or ethanol extract (EARMG) of ARMG (125, 250 or 500 mg/kg) or aspirin (30 mg/kg, a reference control) for 3 weeks. Forty min after the final administration, carotid arterial thrombosis was induced by applying a 70% FeCl₃-soaked filter paper outside the arterial wall for 5 min, and the blood flow was monitored with a laser Doppler probe. Both PARMG and EARMG delayed the FeCl₃-induced arterial occlusion in a dose-dependent manner, doubling the occlusion time at high doses. In mechanism studies, a high concentration of EARMG inhibited platelet aggregation induced by collagen in vitro. In addition, EARMG improved the blood lipid profiles, decreasing triglyceride and cholesterol levels. Although additional action mechanisms remain to be clarified, it is suggested that ARMG containing high amount of ginsenosides such as Rg₃ improves blood flow not only by inhibiting oxidative thrombosis, but also by modifying blood lipid profiles.

Preliminary Study on Adventitious Root Induction and Culturing ofAconitum Pendulum Busch / 中国中医药信息杂志

Objective To establish the adventitious root induction and culture system ofAconitum pendulum Busch.Methods The influence of different explants and plant growth substances on adventitious induction proliferation ofAconitum pendulumBusch were investigated through tissue culturing.Results Explants induction ability was: root>stem>leaf. 2.5 mg/L IBA was in favor of induction and multiplication adventitious root ofAconitum pendulumBusch.ConclusionThe culture system of induction and proliferation adventitious roots ofAconitum pendulum Busch was preliminary established, which provides a new way for resource exploitation and utilization of Aconitum pendulum Busch.

Induction and proliferation culture of Adventitious roots of Aconitum pendulum Busch / 中国药学杂志

OBJECTIVE: To establish the induction and culture system of adventitious roots of Aconitum pendulum Busch. METHODS: By tissue culture, the influences of different explants and plant growth substances on the induction and proliferation of adventitious roots of Aconitum pendulum Busch were investigated. RESULTS: Mixture of MS+2.5 mg·L-1 indole-3-butyric acid (IBA)+0.5 mg·L-1 6-benzylaminopurine (6-BA)+1.0 g·L-1 polyvinyl pyrrolidone (PVP) was the best callus induction medium. Mixture of MS+0.5 mg·L-1 IBA+1.0 g·L-1 PVP was the optimum adventitious root induction medium. Mixture of MS+0.5 mg·L-1 IBA+1.0 g·L-1 PVP was the most suitable adventitious root proliferation medium. CONCLUSION: A culture system is preliminarily established for the induction and proliferation of adventitious roots of Aconitum pendulum, which provides a new way for the resource exploitation and utilization of Aconitum pendulum Busch.

In vitro culture of petiole longitudinal thin cell layer explants of vietnamese ginseng (PANAX VIETNAMENSIS HA ET GRUSHV.) and preliminary analysis of saponin content.

The present work describes a procedure that allows for the easy and rapid induction of somatic embryos, calli, shoots and adventitious roots of Vietnamese ginseng (Panax vietnamensis Ha et Grushv.) from longitudinal thin cell layers (lTCLs). In order to investigate the morphogenesis of this medicinal plant, the effect of separately–supplemented plant growth regulators and combinatorial effect of co–supplemented auxins and cytokinins in dark or under 16-hour photoperiod was examined. After eight weeks of culture, the lTCL explants excised from petiole of three-month-old in vitro plants and cultured on a semi solid basal Murashige and Skoog (MS) media supplemented with 1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.1 mg/l thidiazuron (TDZ) in dark, 2.0 mg/l α-napththaleneacetic acid (NAA) in dark and 1.0 mg/l 2,4-D under light gave the highest rate of callogenesis (100%), embryogenesis (53.3%) and adventitious root formation (100% with a mean of 16.7 roots), and shoot formation (26.7%), respectively. The metabolite of petiole lTCL-derived calli qualitative and quantitative analyses were performed by using high-performance liquid chromatography and thin layer chromatography. The simple procedure, together with similar saponin profiles between the resulted in vitro tissues and plants grown in nature, suggest its potential use in generating Vietnamese ginseng in large amount for medicinal purpose.

Improvement of Growth and Periplocin Yield of Periploca sepium Adventitious Root Cultures by Altering Nitrogen Source Supply / 中草药·英文版

Objective To increase the ultimate yield of periplocin in Periploca sepium adventitious root cultures by a two-stage culture based on nitrogen source.Methods Firstly,the effects of nitrogen source(NH-NO-)at different ratios and different total initial nitrogen amounts on the accumulation of biomass and secondary metabolites in adventitious root cultures of P sepium were investigated,and growth and production media for the two-stage culture based on the above results were established.Results The highest biomass and periplocin content were obtained in the culture medium of 15 mmol/L total nitrogen amount with NH-NO(1:2)and 30 mmol/L total nitrogen amount with nitrate as the sole nitrogen source.By adopting a fed-batch cultivation strategy,the dry weight adventitious root,periplocin content and yield were increased by 136%,108%,and 389%,respectively when compared with those of the control,reaching up to 8.13 g/L,157.15 μg/g,and 1277.63 μg/L,respectively.Furthermore,it was found that in the process of two-stage culture,the adventitious roots grew thicker significantly after they were transferred into production medium directly.Conclusion The ultimate yield of periplocin in P.sepium adventitious root cultures could be significantly increased by a two-stage culture based on nitrogen source.