scholarly journals Chitosan Elicitation Impacts Flavonolignan Biosynthesis in Silybum marianum (L.) Gaertn Cell Suspension and Enhances Antioxidant and Anti-Inflammatory Activities of Cell Extracts

Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 791
Author(s):  
Muzamil Shah ◽  
Hasnain Jan ◽  
Samantha Drouet ◽  
Duangjai Tungmunnithum ◽  
Jafir Hussain Shirazi ◽  
...  
Keyword(s):  
Cell Suspension ◽  
Biomass Production ◽  
Suspension Cultures ◽  
Total Phenolic ◽  
Naphthalene Acetic Acid ◽  
Silybum Marianum ◽  
Enhanced Production ◽  
Cell Extracts ◽  
Anti Inflammatory

Silybum marianum (L.) Gaertn is a rich source of antioxidants and anti-inflammatory flavonolignans with great potential for use in pharmaceutical and cosmetic products. Its biotechnological production using in vitro culture system has been proposed. Chitosan is a well-known elicitor that strongly affects both secondary metabolites and biomass production by plants. The effect of chitosan on S. marianum cell suspension is not known yet. In the present study, suspension cultures of S. marianum were exploited for their in vitro potential to produce bioactive flavonolignans in the presence of chitosan. Established cell suspension cultures were maintained on the same hormonal media supplemented with 0.5 mg/L BAP (6-benzylaminopurine) and 1.0 mg/L NAA (α-naphthalene acetic acid) under photoperiod 16/8 h (light/dark) and exposed to various treatments of chitosan (ranging from 0.5 to 50.0 mg/L). The highest biomass production was observed for cell suspension treated with 5.0 mg/L chitosan, resulting in 123.3 ± 1.7 g/L fresh weight (FW) and 17.7 ± 0.5 g/L dry weight (DW) productions. All chitosan treatments resulted in an overall increase in the accumulation of total flavonoids (5.0 ± 0.1 mg/g DW for 5.0 mg/L chitosan), total phenolic compounds (11.0 ± 0.2 mg/g DW for 0.5 mg/L chitosan) and silymarin (9.9 ± 0.5 mg/g DW for 0.5 mg/L chitosan). In particular, higher accumulation levels of silybin B (6.3 ± 0.2 mg/g DW), silybin A (1.2 ± 0.1 mg/g DW) and silydianin (1.0 ± 0.0 mg/g DW) were recorded for 0.5 mg/L chitosan. The corresponding extracts displayed enhanced antioxidant and anti-inflammatory capacities: in particular, high ABTS antioxidant activity (741.5 ± 4.4 μM Trolox C equivalent antioxidant capacity) was recorded in extracts obtained in presence of 0.5 mg/L of chitosan, whereas highest inhibitions of cyclooxygenase 2 (COX-2, 30.5 ± 1.3 %), secretory phospholipase A2 (sPLA2, 33.9 ± 1.3 %) and 15-lipoxygenase (15-LOX-2, 31.6 ± 1.2 %) enzymes involved in inflammation process were measured in extracts obtained in the presence of 5.0 mg/L of chitosan. Taken together, these results highlight the high potential of the chitosan elicitation in the S. marianum cell suspension for enhanced production of antioxidant and anti-inflammatory silymarin-rich extracts.

scholarly journals Chitosan Elicitation Impacts Flavonolignan Biosynthesis in Silybum marianum (L.) Gaertn Cell Suspension and Enhances Antioxidant and Anti-inflammatory Activities of Cell Extracts

2021 ◽  
Author(s):  
Muzamil Shah ◽  
Hasnain Jan ◽  
Samantha Drouet ◽  
Duangjai Tungmunnithum ◽  
Jafir Hussain Shirazi ◽  
...  
Keyword(s):  
Cell Suspension ◽  
Biomass Production ◽  
Naphthalene Acetic Acid ◽  
Silybum Marianum ◽  
Biotechnological Production ◽  
High Accumulation ◽  
Enhanced Production ◽  
Cell Extracts ◽  
Anti Inflammatory

Silybum marianum (L.) Gaertn is a rich source of antioxidants and anti-inflammatory flavonolignans with great potential for use in pharmaceutical and cosmetic products. Its biotechnological production using in vitro culture system has been proposed. Chitosan is a well-known elicitor that strongly affects both secondary metabolites and biomass production by plants. The effect of chitosan on S. marianum cell suspension is not known yet. In the present study, suspension cultures of S marianum were exploited for their in vitro potency to produce bioactive flavonolignans in the presence of chitosan. Established cell suspension culture was maintained on the same hormonal media supplemented with 0.5 mg/L BAP (6-benzylaminopurine) and 1.0 mg/L NAA (α-naphthalene acetic acid) under photoperiod 16/8 h (light/dark) and exposed to various treatments of chitosan (ranging from 0.5 to 50.0 mg/L). The highest biomass production was observed for cell suspension treated with 5.0 mg/L chitosan, resulting in 123.3 g/L fresh weight (FW) and 17.7 g/L dry weight (DW) productions. Chitosan treatment resulted in an overall increase in the accumulation of flavonoids, phenolic compounds and silymarin. High accumulation levels of silybin B, silydianin and silybin A were recorded by HPLC analysis. The corresponding extracts displayed interesting antioxidant and anti-inflammatory capacities. In particular, high ABTS antioxidant activity (741.5 μM Trolox C equivalent antioxidant capacity) was recorded in extracts obtained in presence of 0.5 mg/L of chitosan. On the opposite, highest inhibitions of cyclooxygenase 2 (COX-2, 30.5 %), secretory phospholipase A2 (sPLA2, 33.9 %) and 15-lipoxygenase (15-LOX-2, 31.6 %) enzymes involved in inflammation process were measured in extracts obtained in presence of 5.0 mg/L of chitosan. Taken together, these results highlight the high potential of the chitosan elicitation of the S. marianum cell suspension for enhanced production of antioxidant and anti-inflammatory silymarin-rich extracts.

scholarly journals Interactive Effects of Light and Melatonin on Biosynthesis of Silymarin and Anti-inflammatory Potential in Callus Cultures of Silybum marianum L.

2019 ◽  
Author(s):  
Muzamil Shah ◽  
Muhammad Asad Ullah ◽  
Samantha Drouet ◽  
Muhammad Younas ◽  
Duangjai Tungmunnithum ◽  
...  
Keyword(s):  
Continuous Light ◽  
Biomass Accumulation ◽  
Interactive Effects ◽  
Total Phenolic ◽  
Naphthalene Acetic Acid ◽  
Phytochemical Analysis ◽  
Silybum Marianum ◽  
Light Regimes ◽  
Anti Inflammatory

Silybum marianum L. is a well-known medicinal herb, primarily used in liver protection. Light strongly affects several physiological processes along with secondary metabolites biosynthesis in plants. Herein, S. marianum was exploited for in vitro potential under different light regimes in the presence of melatonin. The optimum callogenic response occurred in combination of 1.0 mg/L α-naphthalene acetic acid and 0.5 mg/L 6-Benzylaminopurine under photoperiod. Continuous light associated with melatonin treatment increased total flavonoid content (TFC), total phenolic content (TPC) and antioxidant potential, followed by photoperiod and dark treatments. The increased level of melatonin has a synergistic effect on biomass accumulation under continuous light and photoperiod, while adverse effect was observed under dark condition. More detailed phytochemical analysis showed maximum total silymarin content (11.92 mg/g DW) when placed under continuous light + 1.0 mg/L melatonin. Individually, the level of silybins (A and B), silydianin, isolsilychristin and silychristin was found highest under continuous light. Anti-inflammatory activities were also studied and highest percent inhibition was recorded against 15-LOX for cultures cultivated under continuous light (42.33%). The current study helps to better understand the influence of melatonin and different light regimes on silymarin production as well as antioxidant and anti-inflammatory activities in S. marianum callus extracts.

scholarly journals Interactive Effects of Light and Melatonin on Biosynthesis of Silymarin and Anti-Inflammatory Potential in Callus Cultures of Silybum marianum (L.) Gaertn.

Molecules ◽  
2019 ◽  
Vol 24 (7) ◽  
pp. 1207 ◽  
Author(s):  
Muzamil Shah ◽  
Muhammad Ullah ◽  
Samantha Drouet ◽  
Muhammad Younas ◽  
Duangjai Tungmunnithum ◽  
...  
Keyword(s):  
Continuous Light ◽  
Dry Weight ◽  
Interactive Effects ◽  
Total Phenolic ◽  
Naphthalene Acetic Acid ◽  
Phytochemical Analysis ◽  
Silybum Marianum ◽  
Light Regimes ◽  
Anti Inflammatory

Silybum marianum (L.) Gaertn. is a well-known medicinal herb, primarily used in liver protection. Light strongly affects several physiological processes along with secondary metabolites biosynthesis in plants. Herein, S. marianum was exploited for in vitro potential under different light regimes in the presence of melatonin. The optimal callogenic response occurred in the combination of 1.0 mg/L α-naphthalene acetic acid and 0.5 mg/L 6-benzylaminopurine under photoperiod. Continuous light associated with melatonin treatment increased total flavonoid content (TFC), total phenolic content (TPC) and antioxidant potential, followed by photoperiod and dark treatments. The increased level of melatonin has a synergistic effect on biomass accumulation under continuous light and photoperiod, while an adverse effect was observed under dark conditions. More detailed phytochemical analysis showed maximum total silymarin content (11.92 mg/g dry weight (DW)) when placed under continuous light + 1.0 mg/L melatonin. Individually, the level of silybins (A and B), silydianin, isolsilychristin and silychristin was found highest under continuous light. Anti-inflammatory activities were also studied and highest percent inhibition was recorded against 15-lipoxygenase (15-LOX) for cultures cultivated under continuous light (42.33%). The current study helps us to better understand the influence of melatonin and different light regimes on silymarin production as well as antioxidant and anti-inflammatory activities in S. marianum callus extracts.

scholarly journals Salycilic Acid Induces Exudation of Crocin and Phenolics in Saffron Suspension-Cultured Cells

Plants ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 949 ◽  
Author(s):  
Azar Moradi ◽  
Fatemeh Zarinkamar ◽  
Stefania De Domenico ◽  
Giovanni Mita ◽  
Gian Pietro Di Sansebastiano ◽  
...  
Keyword(s):  
Cell Growth ◽  
Cell Suspension ◽  
Cultured Cells ◽  
Water Soluble ◽  
Crocus Sativus ◽  
Stress Effect ◽  
Metabolite Production ◽  
Enhanced Production ◽  
Cell Extracts

The production of crocin, an uncommon and valuable apocarotenoid with strong biological activity, was obtained in a cell suspension culture of saffron (Crocus sativus L.) established from style-derived calli to obtain an in-vitro system for metabolite production. Salycilic acid (SA) was used at different concentrations to elicit metabolite production, and its effect was analyzed after a 4 days of treatment. HPLC-DAD analysis was used for total crocin quantification while the Folin-Ciocâlteu method was applied for phenolic compounds (PC) content. Interestingly, despite cell growth inhibition, a considerable exudation was observed when the highest SA concentration was applied, leading to a 7-fold enhanced production of crocin and a 4-fold increase of phenolics compared to mock cells. The maximum antioxidant activity of cell extracts was evidenced after SA 0.1 mM elicitation. Water-soluble extracts of saffron cells at concentrations of 1, 0.5, and 0.1 µg mL−1 showed significant inhibitory effects on MDA-MB-231 cancer cell viability. The heterologous vacuolar markers RFP-SYP51, GFPgl133Chi, and AleuRFP, were transiently expressed in protoplasts derived from the saffron cell suspensions, revealing that SA application caused a rapid stress effect, leading to cell death. Cell suspension elicitation with SA on the 7th day of the cell growth cycle and 24 h harvest time was optimized to exploit these cells for the highest increase of metabolite production in saffron cells.

scholarly journals Rosmarinic acid production in cell suspension cultures of Ehretia asperula Zollinger & Moritz

2022 ◽  
Vol 9 (1) ◽  
pp. 70-75
Author(s):  
Pham Thi My Tram ◽  
Ngo Ke Suong ◽  
Le Thi Thuy Tien
Keyword(s):  
Cell Suspension ◽  
Rosmarinic Acid ◽  
Rotation Speed ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Inoculum Size ◽  
Naphthalene Acetic Acid ◽  
Callus Line ◽  
Pharmaceutical Industries

Plant cell cultures provide an alternative means for producing secondary compounds in food, cosmetic and pharmaceutical industries. Ehretia asperula Zollinger & Moritzi is used as a traditional medicine for the treatment of liver detoxification, ulcers, tumors, inflammation and enhancing the body's resistance in Vietnam. The study was carried out to select suitable callus line for cell suspension cultures of E. asperula Zollinger & Moritzi and investigate the effects of inoculum size, rotation speed and naphthalene acetic acid (NAA) on the proliferation of cell suspension cultures. In addition, the influence of light intensity on the growth and rosmarinic acid (RA) biosynthesis of cell suspension was also surveyed. After 4 weeks of culture, the white to pale yellow friable callus expanded significantly with a fresh weight (FW) of 0.788 g and a high RA content of 2.062 mg/g FW. An appropriate medium for cell proliferation was the liquid B5 medium, which contained 30 g/l glucose, 0.1 mg/l benzyl adenine (BA) and 0.4 mg/l NAA. The results also demonstrated that a 1:20 ratio (w/v) inoculum size, darkness and rotation speed of 90 rpm were the optimal conditions for the proliferation and RA accumulation to 188.217 mg/l in 4 weeks of culture. These findings showed that E. asperula Zollinger & Moritzi cell suspension cultures could be a potential alternative approach for RA production in vitro.

scholarly journals Antimicrobial activity of callus and cell suspension cultures extracts of Thevetia peruviana

2019 ◽  
Vol 8 (1) ◽  
pp. 45-56
Author(s):  
Juan Pablo Arias Echeverri ◽  
Isabel Cristina Ortega ◽  
Mariana Peñuela ◽  
Mario Arias
Keyword(s):  
Antimicrobial Activity ◽  
Cell Suspension ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Ornamental Plant ◽  
Thevetia Peruviana ◽  
Hexane Extracts ◽  
Ethanol Extracts ◽  
Different Parts

Thevetia peruviana is an ornamental plant considered source of biologically compounds with cardiac and antimicrobial activity. These compounds are normally extracted from different parts of the fully growth plants. In this work, extracts were obtained from callus and cell suspension cultures of T. peruviana and their antimicrobial activity was evaluated by disk diffusion tests against gram negative (Salmonella thipimurium and Escherichia coli) and gram positive (Staphylococcus aureus and Bacillus cereus) strains. Ethanol, methanol and hexane extracts from callus and cell suspension cultures showed biological activity. Methanolic cell suspension extract showed activity against B. cereus and S. aureus. Ethanolic cell suspension extract inhibit all the bacteria, especially S. thipimurium while hexanic extract showed resistance activity against S. thipimurium, S. aureus and B. cereus. In terms of the source of the extracts, hexane extracts obtained from cell suspension cultures showed a higher antimicrobial activity compared to callus, while ethanol extracts had an inverse behavior. These results outline in vitro cell culture of T. peruviana as a feasible biotechnological platform for the production of compounds with antimicrobial activity.

Über den Abbau von Flavonolen in pflanzlichen Zellsuspensionskulturen / Degradation of Flavonols in Plant Suspension Cultures

1972 ◽  
Vol 27 (8) ◽  
pp. 946-954 ◽  
Author(s):  
Wolfgang Hösel ◽  
Paul D. Shaw ◽  
Wolfgang Barz
Keyword(s):  
Salicylic Acid ◽  
Glycine Max ◽  
Cell Suspension ◽  
Cicer Arietinum ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Enzyme Preparations ◽  
Cicer Arietinum L

The flavonols kaempferol, quercetin and isorhamnetin were labelled with 14C by keeping seven day old Cicer arietinum L. plants in an atmosphere of 14CO2 for five days. The purified (U-14C) flavonols were applied to cell suspension cultures of Cicer arietinum L., Phaseolus aureus Roxb., Glycine max and Petroselinum hortense. Based on the rates of 14CO2 formation and distribution of radioactivity after fractionation of the cells, the flavonols were shown to be catabolized to a very high extent.All four cell suspension cultures possess the enzymatic activity transforming flavonols to the recently discovered 2,3-dihydroxyflavanones. Upon incubation of the flavonols datiscetin and kaempferol with enzyme preparations from Cicer arietinum L. cell suspension cultures, it was demonstrated that the enzymatically formed 2,3-dihydroxyflavanones are further transformed in an enzyme catalyzed reaction. Salicylic acid was found as a degradation fragment of ring B of the 2,3,5,7,2′-pentahydroxyflavanone derived from datiscetin. Neither phloroglucinol nor phloroglucinol carboxylic acid were observed as metabolites of ring A. These in vitro findings were further substantiated by in vivo data because the flavonols kaempferol, quercetin and datiscetin when applied to cell suspension cultures of Cicer arietinum L. and Glycine max gave rise to para-hydroxybenzoic acid, protocatechuic acid and salicylic acid, respectively. It was thus concluded that flavonols are catabolized via 2,3-dihydroxyflavanones with the B-ring liberated as the respective benzoic acid. The data are discussed in connection with earlier findings on the catabolism of chalcones, cinnamic and benzoic acids.

scholarly journals Extracts from Cell Suspension Cultures of Strawberry (Fragaria x ananassa Duch): Cytotoxic Effects on Human Cancer Cells

Molecules ◽  
2019 ◽  
Vol 24 (9) ◽  
pp. 1738 ◽  
Author(s):  
Simona Lucioli ◽  
Fabio Pastorino ◽  
Paolo Nota ◽  
Giulia Ballan ◽  
Andrea Frattarelli ◽  
...  
Keyword(s):  
Cell Suspension ◽  
Cell Lines ◽  
Human Cancer ◽  
Mass Measurement ◽  
Human Fibroblasts ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Chromatographic Retention ◽  
Cervix Carcinoma

Natural compounds are emerging as agents for the treatment of malignant diseases. We previously showed that extracts from in vitro cell suspension cultures of strawberry reduced murine melanoma cell proliferation, as shown for fruit extracts. In this work, chromatographic, mass spectrometric, and spectrophotometric analyses were carried out to identify the bioactive compound exerting the detected cytotoxic activity. Moreover, aiming to confirm the anti-proliferative activity of the extracts against both paediatric and adult human tumors, cytotoxic experiments were performed on neuroblastoma, colon, and cervix carcinoma cell lines. Extracts from in vitro cell suspension cultures of strawberry induced a statistically significant reduction of cell growth in all the tumor cell lines tested. Interestingly, human fibroblasts from healthy donors were not subjected to this cytotoxic effect, highlighting the importance of further preclinical investigations. The accurate mass measurement, fragmentation patterns, and characteristic mass spectra and mass losses, together with the differences in chromatographic retention times and absorbance spectra, led us to hypothesize that the compound acting as an anti-proliferative agent could be a novel acetal dihydrofurofuran derivative (C8H10O3, molecular mass 154.0630 amu)

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