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.

Effect of methyl jasmonate on salidroside and polysaccharide accumulation in Rhodiola sachalinensis callus / 中国中药杂志

<p><b>OBJECTIVE</b>To provide a new material for producing the Rhodiolasachalinensis products, the effect of methyl jasmonate (MeJA) on callus biomass and effective compound accumulation of Rhodiolasachalinensis was studied.</p><p><b>METHOD</b>The calluses-cultured in 3 L-air lift balloon type bioreactor were treated with MeJA after 20 d of bioreactor culture and the effect of MeJA concentration and treatment days on callus biomass, salidroside or polysaccharide accumulation and superoxide dismutase (SOD) and peroxidase (POD) activities were investigated.</p><p><b>RESULT</b>The callus biomass was not significantly different after MeJA treatment (125) for 0-6 d but obviously decreased after 6 d treatment. The maximum salidroside or polysaccharide contents and SOD or POD activities were found after 4 d treatment of MeJA. MeJA concentration significantly affected callus biomass and effective compound accumulation, biomass decreased at MeJA concentrations higher than 125 μmol x L(-1). However, the effective compound contents were determined at higher MeJA concentration, and the highest salidroside and polysaccharide accumulation was found at 225 and 275 μmol x L(-1) MeJA, respectively and the maximum SOD and POD activities was found at 225 μmol x L(-1) MeJA. The effective compound contents in callus were compared with field-grown plants. Salidroside contents in calluses were 1.1-fold and 2. 4-fold more than in plant roots and stem or leave, respectively. Polysaccharide content in calluses were 3. 6-fold and 8.0-fold more than in plant roots and stem or leave, respectively.</p><p><b>CONCLUSION</b>Salidorside and polysaccharide in Rhodiolasachalinensiscalluses improved by MeJA treatment, 225 μmol x L(-1) MeJA and 4 d treatment were optimal. The effective compound contents in callus were obviously higher than in field-grown plants. Therefore, bioreactor culture is efficient for obtaining mass effective compounds of Rhodiolasachalinensis by culturing calluses. This method could provide an alternative material source for production of Rhodiolasachalinensis products.</p>

Effects of several factors on cell growth and ginsenoside accumulation of Panax ginseng suspension culture / 中国中药杂志

To improve cell suspension culture system of Panax ginseng, the dynamic of cell growth and medium consumption were studied, and the effects of filter on the culture vessel, revolution number, and inoculation density on cell growth and ginsenoside accumulation were also investigated. The maximum cell growth and ginsenoside accumulation was found on the 20th days of suspension culture, therefore, 20 days were confirmed as a suitable culture period for mass production of ginsenoside. Cell growth and ginsenoside content were promoted when the culture vessel had a ventilated filter. Revolution speed during suspension culture affected cell growth, but not ginsenoside content, a peak of ginsenoside productivity was found in the treatment of 120 r x min(-1). Inoculation density also influenced cell growth and ginsenoside accumulation, inoculation density of 6 g was better than other inoculation densities, the ginsenoside content and productivity were up to 12.8 mg x g(-1) DW and 146.6 mg x L(-1), respectively.

Effects of acid and alkali stress on ginsenoside content and histochemical localization of ginsenoside in adventitious root of Panax ginseng / 中国中药杂志

To investigate the effect of acid and alkali stress on ginsenoside content of Panax ginseng, adventitious roots culture in bioreactors were incubated for 30 d and pH value was adjusted. Ginsenoside content increased by reducing or raising the pH in culture medium, the muxium ginsenoside content was determined on the 5th days after acid treatment and on the 7th days after alkali treatment. The result of histochemical localization of ginsenoside revealed that the red color from light to dark were found in the adventitious root tissue, and ginsenoside mainly located in the pericycle cells where appeared the dark red color.

Cultivation of protocorms of Dendrobium candidum in air-lift bioreactors / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the factors affecting the growth of protocorms of Dendrobium candidum and substance synthesis in a reactor, in order to provide a new method for mass production of raw materials of D. candidum.</p><p><b>METHOD</b>Protocorms in vitro were used as experimental materials to study the effect of inoculum volume, light intensity and air volume on the growth of protocorms of D. candidum and the accumulation of polysaccharide and dendrobine in a 3 L-air lift balloon type bioreactor.</p><p><b>RESULT</b>After 30 days of cultivation in a bioreactor, protocorms became dark green and grew well at the inoculum volume of 10 g x L(-1). The polysaccharide content in protocorms showed no difference at various inoculum volumes; whereas the dendrobine content showed differences (with the highest treatment at the inoculum volume of 10 g x L(-1)), particularly the productions of polysaccharide and alkaloid reached the maximum at the inoculum volume of 10 g x L(-1). The condition of 1 600 lx of light intensity was the most favorable for the growth of protocorms. Though light played a role of improving the accumulation of polysaccharide in protocorms of D. candidum, it could inhibit the accumulation of dendrobine. Polysaccharide content and production were better under light conditions of 1 600 and 2 400 lx than dark conditions. Despite the maximum dendrobine content in dark conditions, the dendrobine production showed the maximum in the light condition of 1 600 lx due to poor growth of protocorms. Protocorms grew well and became dark green at the air volume of 0.2 vvm (air volume culture volume per minute) , which was better than at 0.1 and 0.3, with maximum polysaccharide and dendrobine contents and productions.</p><p><b>CONCLUSION</b>In a 3 L-air lift balloon type bioreactor with a working volume of 2 L, the conditions of 10 inoculum volume, 1 600 lx light intensity and 0.2 air volume were favorable for the growth of protocorms and the production of dendrobine. This demonstrates that the cultivation of D. candidum and substance synthesis in a reactor is an effectie approach for mass production of polysaccharide and dendrobine.</p>

Preliminary study on cultivation of adventitious roots of Hypericum perforatum in bioreactors / 中国中药杂志

<p><b>OBJECTIVE</b>To cultivate adventitious roots of Hypericum perforatum in bioreactors, in order to seek for suitable conditions for adventitious growth.</p><p><b>METHOD</b>The effect of IBA concentration, sugar type and concentration, inoculum volume and air volume of adventitious roots on the cultivation of adventitious roots of H. perforatum was observed in a 5 L air-lift bioreactor.</p><p><b>RESULT</b>Adventitious roots of H. perforatum were cultivated in a MS culture dish. With the increase of IBA concentration, the propagation coefficient of adventitious roots of H. perforatum was on the rise. The IBA concentration ranging between 1.25-1.75 mg x L(-1) was suitable for the growth of adventitious roots. Adventitious roots grew best with sucrose in MS medium, with the propagation coefficient up to 22.15. When sucrose concentration was 30 g x L(-1), fresh weight, dry weight and propagation coefficient reached the maximum value. An adventitious root reactor with an inoculum volume of 20 g was favorable for the growth of adventitious roots. The air volume of reactors of 0.075 vvm (air volume/culture volume per minute) was favorable for the growth of adventitious roots, with the significant increase in the propagation coefficient of adventitious roots. In the amplification experiment, we found that the cultivation conditions of adventitious roots in a 5 L bioreactor was completely applicable to that in 10 and 20 L bioreactors, and adventitious roots grew well in a large bioreactor.</p><p><b>CONCLUSION</b>IBA concentration, sugar type and concentration, inoculum volume and air volume had a significant effect on the growth of adventitious roots.</p>