Highest Rosmarinic Acid Content Research Articles

Elicitation with multi-walled carbon nanotubes improved growth and production of polyphenolic compounds in Melissa officinalis

Carbon nanotubes as novel plant elicitors are intensively studied in biotechnology due to their concentration-dependent effects on plant health. This emphasizes the importance of studying nanomaterials in the field of plant nanotoxicology and enables a better understanding of their advantages and disadvantages for plant health. Researchers examined how various concentrations of multi-walled carbon nanotubes (MWCNTs) affect growth and polyphenolic accumulation in the medicinal herb, Melissa officinalis. Two-month-old plant shoots were sprayed with various concentrations (0–250 mg L−1) of COOH- functionalized MWCNTs and harvested two and three weeks after elicitation. TEM images confirmed MWCNTs uptake into the mesophyll and the vessels of leaves. Low to moderate MWCNT concentrations (50–100 mg L−1) boosted growth indices and increased total amount of phenols, flavonoids, and phenolic acids, peaking three weeks after treatment with 100 mg L−1 MWCNTs, without destroying the cells and subcellular organelles. HPLC analysis showed this treatment yielded the highest content of rosmarinic acid, salvianolic acid B, apigenin, and kaempferol. However, 250 mg L−1 MWCNTs damaged cells without boosting metabolite production. We suggest that elicitation with low to moderate MWCNT concentrations can be a useful tool for laboratory-scale production of phenolic metabolites in M. officinalis.

Exploring the impact of exogenous melatonin on agro-morphological characteristics, carvacrol, and rosmarinic acid production in Satureja rechingeri Jamzad under drought stress

Satureja rechingeri Jamzad (known as “Jatra” in Persian), which belongs to the Lamiaceae family, is a rich source of essential oil particularly carvacrol, and rosmarinic acid. Drought stress has a detrimental impact on the physiological and biochemical parameters of plants, leading to a decline in plant productivity. Melatonin (MT), a new plant growth regulator found abundantly in plants, has been found to enhance the plant's internal resistance to various environmental stresses. The present study aimed to examine the impact of exogenously applied MT on the agro-morphological, physio-biochemical, and phytochemical traits of S. rechingeri plants cultivated under different levels of drought stress. The results indicated that plants treated with 200 µM MT obtained the highest plant height, length and width of leaf, fresh, dry and drug weight under different drought stress levels. The highest values of relative water content (RWC) (93.5%) and chlorophyll content (15.4 mg/g FW) were recorded by MT 200 µM and 100 µM, respectively, in 100% FC. Drought stress treatments (40, 60, and 80% FC) without foliar spray of MT significantly enhanced the H2O2 content, electrolyte leakage, and malondialdehyde content in leaves, whereas MT treatment under drought stress significantly decreased the above parameters. The lowest H2O2 content (11.5 nmol/g), electrolyte leakage (3.08%), and malondialdehyde content (0.78 µM/g) were obtained by 200 µM MT at 100% FC. In contrast, drought stress treatment increased the total phenol content (TPC), rosmarinic acid (RA), essential oils (EOs) content and yield, and carvacrol. The maximum values of TPC (28.1 mg GAE/g DW), EOs content (3.63%) and yield (0.96%), and carvacrol (95.66%) were achieved by 200 µM MT at 40% FC. The highest RA content (7.43 mg GAE/g DW) was recorded in 100 µM MT at 40% FC. Thus, foliar spray MT has the potential to enhance plant growth through the mitigation of reactive oxygen species (ROS)-induced oxidative harm, as well as the augmentation of photosynthesis pigments, secondary metabolites such as phenolics, EOs levels, overall antioxidant scavenging capacity, and the preservation of RWC during periods of drought stress.Graphical

Rosmarinic acid-rich ethanolic extract of Orthosiphon stamineus ameliorates cognitive and hippocampal long-term potentiation in chronic cerebral hypoperfusion rat model

Chronic cerebral hypoperfusion (CCH) is one of the main causes of vascular dementia caused by the reduced blood flow to the brain. Orthosiphon stamineus (OS) is a medicinal herb exhibiting pronounced neuroprotective, anti-oxidant and anti-inflammatory activities due to its high rosmarinic acid content. This study investigated the nootropic effect of OS ethanolic extract on cognitive functions in CCH rats. CCH was developed by permanent bilateral occlusion of the common carotid artery (PBOCCA). Passive avoidance task (PAT) and Morris water maze (MWM) test were conducted to evaluate cognitive functions followed by in vivo long-term potentiation (LTP) for assessing neuroplasticity. The rosmarinic acid content of OS ethanolic extract was quantified using a validated HPLC-PDA. Treatment with OS ethanolic extract significantly increased step-through latency in the PAT, decreased escape latency at a low dose of OS extract in the MWM and rescued the LTP impairment at the highest dose in CCH rats. These results strongly support the effectiveness of rosmarinic acid-rich OS extract (5.088 % w/w) in treating pathological vascular dementia caused by CCH.

Trachystemon orientalis (L.) G. Don as a Valuable Source of Rosmarinic Acid: Biological Activities and HPLC Profiles.

Trachystemon orientalis (L.) G. Don, colloquially known in Türkiye as "kaldırık", is an edible plant belonging to the Boraginaceae. This plant has been practiced in traditional medicine for many years for its various therapeutic benefits. The effectiveness and chemical composition of plants can vary depending on their parts, age, and extraction solvent. Therefore, the current study aimed to define the biological activities of various parts and extracts of T. orientalis, which were collected in distinct seasons as young and mature, and investigate the main component responsible for these biological effects. Plant material was collected in different seasons from the northwest of Türkiye. 2,2'-Azinobis-(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activities were investigated to assess antiradical and antioxidant potential of the extracts. Anti-inflammatory activity of the extracts was also tested using human red blood cell membrane stabilizing method. Folin-Ciocalteu test was conducted to determine the total phenolic content. Reverse phase-high performance liquid chromatography-photodiode array detector (RP-HPLC-PDA) analysis was performed. Both methanol and aqueous extracts exhibited significant radical scavenging and anti-inflammatory activities compared with control (p<0.05). The highest percentage of inhibition on ABTS and DPPH free radicals was obtained in aqueous extracts of the mature herbs and roots, respectively. Methanol extracts of the mature roots and herbs exhibited the strongest anti-inflammatory capacity. Rosmarinic acid possessed a much higher antioxidant and anti-inflammatory effect than the reference compounds used in each assay in our study. High rosmarinic acid content of the extracts suggests that the compound responsible for the great biological activity potential is rosmarinic acid. To the best of our knowledge, the presence of rosmarinic acid in herbs and roots of T. orientalis was shown for the first time in our present study. Phytochemical composition and effective biological activities of T. orientalis explain its traditional use and indicate its significant potential in pharmaceutical industry applications.

Application of Microwave-Assisted Drying to Shorten Granules Drying Process for the Preparation of Thunbergia laurifolia Lindl. Leaf Tablets

Thunbergia laurifolia Lindl. is a medicinal plant that belongs to the family of Acanthaceae. It possesses several biological and pharmacological activities. According to the Thai National List of Essential Medicines, T. laurifolia leaves are used for the treatment of fever. This work sought to optimize microwave-assisted drying of Thunbergia laurifolia Lindl. leaf granules to shorten the granule drying process by the 32 full factorial design. Power was varied from 300, 450 and 600 W for 5, 10 and 15 min, to minimize moisture content and maximize rosmarinic acid content. Results showed that the optimal microwave-assisted drying condition within the design space and control space was the power of 450 W for 13 min. This condition gave the moisture content of 4.77 ± 0.24 % and rosmarinic acid content of 0.0566 ± 0.0075 %. The HPLC method was validated; it showed good linearity, specificity, precision, and accuracy. Furthermore, the system suitability was achieved. The T. laurifolia leaf granules obtained from the optimal microwave-assisted drying condition were further used to prepare T. laurifolia leaf tablets by the wet granulation method. Three batches of 650 mg tablets containing 250 mg of T. laurifolia leaf powder were prepared using different compression forces: 500, 1,000 and 1,500 psi. The suitable compression force was 1,500 psi; the tablet formulation had suitable hardness (9.88 ± 0.52 kP), low friability (0.20 %), and a short disintegration time (23.17 ± 0.76 s). Rosmarinic acid marker was dissolved from the tablets for approximately 75 % within 4 h when 0.5 % sodium lauryl sulfate aqueous solution was used as dissolution medium. In conclusion, microwave-assisted drying could be applied in preparing T. laurifolia leaf tablets by shortening the granules drying time. HIGHLIGHTS The 32 full factorial design was applied to optimize microwave-assisted drying of Thunbergia laurifolia leaf granules The optimal microwave-assisted drying condition providing the low moisture content with high rosmarinic acid content was the power of 450 W for 13 min The tablets prepared from laurifolia leaf granules had suitable hardness, low friability, short disintegration time, and a high percentage of rosmarinic acid marker dissolved GRAPHICAL ABSTRACT

Detoxification of comfrey (Symphytum officinale L.) extract using natural deep eutectic solvent (NADES) and evaluation of its anti-inflammatory, antioxidant, and hepatoprotective properties.

Comfrey (Symphytum officinale L.) contains rosmarinic acid which has different pharmacological activities, such as antioxidant and anti-inflammatory activities. However, the medicinal use of comfrey is limited by the hepatotoxic effect of lycopsamine in comfrey, which overshadows the health benefits of rosmarinic acid. Natural deep eutectic solvents (NADES) have a wide range of extraction properties, that provides a new approach to the detoxification of comfrey. In the present study, betaine-based and choline chloride-based NADES were screened for selective extraction of rosmarinic acid over lycopsamine. Ultrasonication was used in conjunction with NADES extraction. The chemical profile of the NADES extracts on antioxidant, anti-inflammatory and hepatotoxic activities were investigated using some chemical reagents. Betaine-urea (1:2 molar ratio, 50% water) obtained the highest content of rosmarinic acid and a low level of lycopsamine (1.934 and 0.018mg/g, respectively). Betaine-urea was also shown to be more effective to extract rosmarinic acid compared to methanol-UAE under the same conditions, which gave lower rosmarinic acid and higher lycopsamine levels (0.007 and 0.031mg/g, respectively). Betaine-urea extracts showed higher antioxidant and anti-inflammatory properties as compared with other NADES extracts, however, had lower hepatotoxic profile. This study recommends the use of betaine-urea to detroxify comfrey to open wider opportunities for the development of comfrey for medicinal use.

Protective effect of rosmarinic acid-rich extract from Trichodesma khasianum Clarke against microbiota dysbiosis in high-fat diet-fed obese mice

Hypertrophy of adipose tissues and dysbiosis are hallmarks of obesity. Although drugs are applied for obesity treatment, side effects limit their use. The anti-obesity capacity of rosmarinic acid (RA) has been documented. Trichodesma khasianum Clarke is an edible RA-rich plant grown in Taiwan. Our previous study found that an 80 % ethanol extract of T. khasianum Clarke leaves (80EETC) ameliorates gastric mucosal damage through its anti-inflammatory, antioxidant, and microbiota modulation abilities. However, the anti-obesity effect of 80EETC remains unclear. Therefore, the objective of this study was to explore the protective effects of low-dose 80EETC (125 mg/kg b.w., 80EETCL) or high-dose 80EETC (250 mg/kg b.w., 80EETCH) on obesity development through gut microbiota modulation in high-fat diet (HFD)-induced C57BL/6 mice. The results showed a high RA content (89.2 ± 7.4 mg/g) in 80EETC. 80EETC administration significantly decreased body weight, body fat ratio, serum lipid levels (TC, TG, and LDL-C), adipose tissue accumulation, malondialdehyde (MDA), and tumor necrosis factor-α (TNF-α) in HFD-fed mice. Furthermore, supplementation with 80EETC reduced the Firmicutes/Bacteroidetes ratio and enhanced the relative abundance of gut microbiota (p_Bacteroidetes, f_Lactobacillus, f_Muribaculaceae, f_Prevotellaceae, g_Lactobacillus, g_Prevotellaceae_NK3B31_group, g_Ruminococcaceae_UCG–013, and g_Ruminococcaceae_UCG–014), which negatively correlated with obesity-related factors such as body weight, energy intake, fat accumulation in adipose tissue, TC, TG, LDL, and MDA. In conclusion, RA-rich 80EETC had a protective effect against obesity development and it has potential in healthy food applications.

A Phytocomplex Obtained from Salvia officinalis by Cell Culture Technology Effectively Controls the Grapevine Downy Mildew Pathogen Plasmopara viticola.

The negative impact of using conventional fungicides in plant disease protection has increased the interest in safer alternatives such as plant secondary metabolites, generally having a better toxicological profile. However, cultivation conditions and plant material strongly affect the quality and quantity of secondary metabolites obtained from field grown plants, limiting the standardization needed for industrial production. Plant cell culture technology can provide highly homogeneous biomasses with specific chemical characteristics. A phytocomplex with high rosmarinic acid content (10.12% w/w) was obtained from a selected cell line of Salvia officinalis and was tested against the grapevine downy mildew pathogen, Plasmopara viticola. Grapevine leaf discs were sprayed with the phytocomplex at 5 g/L and then inoculated with P. viticola sporangia. Sporulation level on each disc was assessed after 7 days with an image processing software. The phytocomplex reduced by 95% the sporulation level compared to the control and was also more effective than rosmarinic acid alone, used at the same concentration found in the phytocomplex. Persistence of the phytocomplex was also assessed: when applied 5 days before inoculation, it reduced by 90% the sporulation level compared to the control. These results highlight the possibility to take advantage of cell culture techniques to produce safer pesticides with high quality standards.

Attenuation of neuroinflammation in microglia cells by extracts with high content of rosmarinic acid from in vitro cultured Melissa officinalis L. cells

Plant cell culture is a biotechnology cultivation method that permit to cultivate plants in a short period of time and to obtain extracts with a high degree of standardization and high safety profile. The aim of our study was to evaluate the anti-inflammatory and neuroprotective activity of a standardized Melissa officinalis L. phytocomplex extract (MD) obtained with an in vitro plant cell culture. The MD has been chemically characterized and the content of total polyphenols was 5.17 ± 0.1 % w/w, with a content of rosmarinic acid (RA), its main constituent, of 4.02 ± 0.1 % w/w. MD was tested in an in vitro model of neuroinflammation, in which microglia cells (BV2) were stimulated with Lipopolysaccharides (LPS; 250 ng/mL) for 24 h and its pharmacological activity was compared with that of RA. MD (10 µg/mL) and RA (0.4 μg/mL) reduced pro-inflammatory factors (NF-kB, HDAC, IL-1ß) in LPS-stimulated BV2 cells and counteracted the toxic effect produced by activated microglia medium on neuronal cells. This work shows the efficacy of MD on reducing microglia-mediated neuroinflammation and promoting neuroprotection, highlighting the innovative use of in vitro plant cell cultures to obtain contaminant-free extracts endowed with marked activity and improved quali-quantitative ratio in the constituents’ content.

Alginate encapsulation of micro-cuttings in endangered Satureja khuzistanica species: a promising method for obtaining genetically stable plants with high rosmarinic acid content

Synthetic seed technology is a suitable approach for the rapid, and uniform mass production of Satureja khuzistanica Jamzad, a valuable medicinal plant of the Lamiaceae family. In this study, the encapsulation of micro-cuttings of S. khuzistanica using sodium alginate (SA) was investigated. In order to determine the best conditions for encapsulation, different concentrations of SA and CaCl2·2H2O, different types of explants and matrix culture media, different types and concentrations of plant growth regulators, and different cold storage conditions of synthetic seeds were tested. Also, the genetic stability of regenerated plants from synthetic seeds using the inter simple sequence repeat (ISSR) molecular marker as well as the content of rosmarinic acid in synthetic seed-derived plants were evaluated. The use of 3% SA and 100 mM CaCl2·2H2O was found to be optimal for gel complexation. Encapsulation of nodal segments with 1/2 Murashige and Skoog (MS) medium containing 2.5 µM 6-Benzylaminopurine resulted in the highest regrowth percentage (72.22%) and regrowth rate (0.173). Pre-culture of nodal segments in 1/2 MS medium containing 5 µM indole-3-butyric acid (IBA) and 0.2% activated charcoal for 10 days and the use of 2.5 µM IBA in the rooting medium resulted in the highest rooting percentage (88.88%) in synthetic seed-derived plants. The highest regrowth percentage (61.11%) and regrowth rate (0.173) of synthetic seeds were obtained using 2.0 µM thidiazuron (TDZ) in MS liquid medium. The placement of germinated seeds on the coco peat substrate resulted in the highest conversion rate (61.11%) of synthetic seeds. Also, storage of encapsulated nodal segments at 4 °C in MS culture medium, compared to cold storage without using MS medium, resulted in better regrowth of synthetic seeds. The highest regrowth percentage (44.44%) and regrowth rate (0.092) for cold-stored synthetic seeds occurred after 2 weeks. The genetic stability testing by ISSR molecular marker showed that synthetic seed-derived plantlets were genetically similar to their mother plants. Also, encapsulated nodal segments and shoot tip-derived plants significantly enhanced the rosmarinic acid content up to 7.77 times that of the natural seed-derived plants. Genetic restoration programs, short-term storage, and germplasm distribution in S. khuzistanica plants could all benefit from the encapsulation regeneration strategy reported here.

Development, Characterization and Incorporation of Alginate-Plant Protein Covered Liposomes Containing Ground Ivy (Glechoma hederacea L.) Extract into Candies.

Ground ivy (Glechoma hederacea L.) has been known as a medicinal plant in folk medicine for generations and, as a member of the Lamiaceae family, is characterized with a high content of rosmarinic acid. The aim of the present study was to formulate delivery systems containing bioactive compounds from ground ivy in encapsulated form and incorporated into candies. Liposomes were examined as the encapsulation systems that were additionally coated with an alginate–plant protein gel to reduce leakage of the incorporated material. Bioactive characterization of the ground ivy extract showed a high content of total phenolics (1186.20 mg GAE/L) and rosmarinic acid (46.04 mg/L). The formulation of liposomes with the high encapsulation efficiency of rosmarinic acid (97.64%), with at least a double bilayer and with polydisperse particle size distribution was achieved. Alginate microparticles reinforced with rice proteins provided the highest encapsulation efficiency for rosmarinic acid (78.16%) and were therefore used for the successful coating of liposomes, as confirmed by FT-IR analysis. Coating liposomes with alginate–rice protein gel provided prolonged controlled release of rosmarinic acid during simulated gastro-intestinal digestion, and the same was noted when they were incorporated into candies.

FLAVONOIDS AND LIGNANS FROM THE LEAVES OF NEPETA MULTIFIDA (LAMIACEAE) AND THEIR BIOAC-TIVITY

A perennial plant Nepeta multifida L. (syn. Schizonepeta multifida (L.) Briq.) is one of the most common species of the Lamiaceae family growing in Eastern Siberia and used in traditional oriental medicine. The chemical composition of N. multifida has not been sufficiently studied. Chromatographic separation of phenolic compounds of N. multifida leaves using column chromatography and preparative HPLC resulted in the isolation of sixteen compounds, including a new flavonoid identified as luteolin-7-O-(3′′,6′′-di-O-acetyl)-β-D-glucopyranoside. Known compounds were O-glycosides of luteolin and apigenin, rosmarinic acid, salvianolic acids A and B, and schizotenuin A. Quantitative analysis of N. multifida leaves by HPLC-UV assay showed the high content of rosmarinic acid (8.36–35.71 mg/g), luteolin-7-O-glucuronide (2.03–14.18 mg/g) and schizotenuin A (5.29–9.56 mg/g). The highest level of phenolic compounds was found in the flowering and fruiting phases. Using Ellman’s spectrophotometric method, it was found that N. multifida leaf extract and some compounds had antiacetylcholinesterase activity, and luteolin glycosides being the most active showed the level of concentration of half-maximal enzyme inhibition (IC50) 29.03–58.36 μg/mL. Thus, as a result of the present study, it was found that the leaves of N. multifida contain various groups of phenolic compounds capable of inhibiting the activity of acetylcholinesterase.

Biochemical parameters and antioxidant property of three Salvia L. taxa endemic in Turkey.

The aim of the present study was to determine and compare the fatty acids, lipid soluble vitamins, sterols, phenolics, and antioxidant capacities of three endemic Salvia L. taxa (S. euphratica var. Montbret & Aucher ex Bentham leiocalycina (Rech. Fil.) Hedge, S. euphratica var. Montbret & Aucher ex Bentham euphratica (Rech. Fil.) Hedge and S. pseudoeuphratica Rech.Fil.) and to evaluate these results systematically. The fatty acid compositions were determined by using gas chromatography, while the lipid soluble vitamins, sterols, and phenolics were determined by using HPLC. Also, the antioxidant capacities of three Salvia taxa were measured in vitro. Palmitic acid was found as major saturated fatty acid while oleic acid, linoleic acid, α-linolenic acid, and erucic acid were found as major unsaturated fatty acids in the present study. It was found that S. euphratica var. euphratica had lower palmitic acid (8.94 ± 0.71%), total saturated fatty acid (19.16 ± 0.15%), and higher unsaturated fatty acid content (82.08 ± 0.52%) than other studied taxa. Furthermore, it was shown that S. euphratica var. euphratica had different 18.3/18:2 (0.36) unsaturated/saturated fatty acid (4.28) ratios. However, this study demonstrated that Salvia taxa had low lipid soluble vitamins, sterol contents. On the other hand, it was shown that Salvia taxa had similar catechin (509.2 ± 4.21 µg/g and 552.2 ± 9.21 µg/g) and vanillic acid amounts (351.2 ± 2.17 µg/g and 396.8 ± 4.1 µg/g) in this study. And also, it was found that Salvia taxa had high rosmarinic acid content while S. euphratica var. leiocalycina had the highest rosmarinic acid content (1480 ± 7.57 µg/g). On the other hand, it was shown that the two ferulic acid contents of S. euphratica varieties were higher (1175 ±/5.21 µgmg-1740.2 ± 4.82 µg/mg) than the ferulic acid content of S. pseudoeuphratica of which was the lowest (19.2 ± 0.97 µg/mg). The present results suggested that the biochemical results guided the morphological studies, and Salvia taxa have a potent antioxidant capacity.

Rosmarinic acid production in cell suspension cultures of Ehretia asperula Zollinger &amp; Moritz

Plant cell cultures provide an alternative means for producing secondary compounds in food, cosmetic and pharmaceutical industries. Ehretia asperula Zollinger &amp; 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 &amp; 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 &amp; Moritzi cell suspension cultures could be a potential alternative approach for RA production in vitro.

The Protective Function and Modification of Secondary Metabolite Accumulation in Response to Light Stress in Dracocephalum forrestii Shoots.

The aim of this work was to determine the effect of stress conditions caused by different light sources, i.e., blue LED (λ = 430 nm), red LED (λ = 670 nm), blue and red LED (70%:30%) and white LED (430–670 nm) on the growth and morphology of cultivated in vitro Dracocephalum forrestii shoot culture. It also examines the effects on bioactive phenolic compound production and photosynthetic pigment content, as well as on antioxidant enzyme activity (CAT, SOD, POD) and antioxidant properties. The most beneficial proliferation effect was observed under white LEDs (7.1 ± 2.1 shoots per explant). The white and blue lights stimulated the highest fresh weight gain, while red light induced the highest dry weight gain. The total phenolic acid content ranged from 13.824 ± 1.181 to 20.018 ± 801 mg g DW−1 depending on light conditions. The highest content of rosmarinic acid was found in the control shoots (cultivated under fluorescent lamps), followed by culture grown under red light. All LED treatments, especially red and blue, increased salvianolic acid B content, and blue increased apigenin p-coumarylrhamnoside biosynthesis. The greatest ferric reduction activity was observed in shoots cultivated under red light, followed by blue; this is associated with the presence of the highest total phenol content, especially phenolic acids. Similarly, the highest DPPH radical scavenging potential was observed under red light followed by blue. This study proves that LEDs have emerged as significant support for directed in vitro propagation, taking advantage of specific stress responses on various light spectra. This study also showed how stress induced by different LED light spectra increases in Dracocephalum forrestii the synthesis of pharmacologically-active compounds. Hence, light stress may turn out to be a simpler alternative to metabolic engineering for improving the production of secondary metabolites of therapeutic value.

Analysis of functional ingredients and composition of Ocimum basilicum

Basil (Ocimum basilicum L) extracts showed significant antioxidant activity and represent a rich source of antioxidants that could contribute to its disease-fighting potential. Basil contains functional ingredients due to its rich content of phenolic compounds, characterized as main carriers of its antioxidant activity. The objective of this study was to determine the influence of the conditions and methods of extraction on chemical composition of basil extracts and also to find similarities or dissimilarities of the extraction procedures in the scope of determined content of functional ingredients. The determination of phenolic compounds (phenolic acids and flavonoids) of 100 basil extracts was performed using HPLC. The concentration of phenolic acids (chlorogenic, p-hydroxybenzoic, caffeic, ferulic, vanillic, cinnamic and rosmarinic) varied from 0.01 to 79.88 mg/kg of dried extract (DE). Concentration of flavonoid components (quercetin, naringenin, rutin) ranged from 0.06 to 72.24 mg/kg of DE. Extraction with dichloromethane (10 min), chloroform (10 min), n-hexane (30 min) and infusion provided high content of rosmarinic acid and quercetin, but also a significant content of chlorogenic acid, vanillic acid, naringenin, cinnamic and ferulic acid. Chemometric analysis indicated the most effective experimental procedure and provided an overview of their grouping in the space of the extracted ingredients.

Biotechnologically Produced Lavandula angustifolia Mill. Extract Rich in Rosmarinic Acid Resolves Psoriasis-Related Inflammation Through Janus Kinase/Signal Transducer and Activator of Transcription Signaling.

Psoriasis is a common skin pathology, characterized by dysregulation of epidermal keratinocyte function attended by persistent inflammation, suggesting that molecules with anti-inflammatory potential may be effective for its management. Rosmarinic acid (RA) is a natural bioactive molecule known to have an anti-inflammatory potential. Here we examined the effect of biotechnologically produced cell suspension extract of Lavandula angustifolia Mill (LV) high in RA content as treatment for psoriasis-associated inflammation in human keratinocytes. Regulatory genes from the nuclear factor kappa B (NF-κB) and Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathways were upregulated upon stimulation with a combination of interferon gamma (IFN-γ), interleukin (IL)-17A and IL-22. We also observed that both LV extract and RA could inhibit JAK2, leading to reduced STAT1 phosphorylation. Further, we demonstrated that LV extract inhibited phosphoinositide 3-kinases (PI3K) and protein kinase B (AKT), which could be implicated in reduced hyperproliferation in keratinocytes. Collectively, these findings indicate that the biotechnologically produced LV extract resolved psoriasis-like inflammation in human keratinocytes by interfering the JAK2/STAT1 signaling pathway and its effectiveness is due to its high content of RA (10%). Hence, both LV extract and pure RA possess the potential to be incorporated in formulations for topical application as therapeutic approach against psoriasis.

In Vitro Cultured Melissa officinalis Cells as Effective Ingredient to Protect Skin against Oxidative Stress, Blue Light, and Infrared Irradiations Damages

Skin is being increasingly exposed to artificial blue light due to the extensive use of electronic devices, which can induce cell oxidative stress, causing signs of early photo aging. The Melissa officinalis phytocomplex is a new standardized cosmetic ingredient obtained by an in vitro plant cell culture with a high content of rosmarinic acid. In this study, we examine the activity of the Melissa officinalis phytocomplex to protect skin against blue light and infrared damages, evaluating the ROS (Radical Oxygen Species) level in keratinocyte cell line from human skin (HaCaT) and Nrf2 (Nuclear factor erythroid 2-related factor 2), elastin, and MMP1 (Matrix Metalloproteinase 1) immunostaining in living human skin explants ex vivo. This phytocomplex demonstrates antioxidant activity by reducing ROS production and thus the oxidant damage of the skin caused by UV and blue light exposure. In addition, it inhibits blue light-induced Nrf2 transcriptional activity, IR-induced elastin alteration, and IR-induced MMP-1 release. This Melissa officinalis phytocomplex is a new innovative active ingredient for cosmetic products that is able to protect skin against light and screen exposure damages and oxidative stress.

Comparative study of callus culture and leaves of Thunbergia laurifolia for their bioactive constituents and the activation of AMPK and GLUT‐dependent glucose uptake on rat skeletal muscle (L6) cells

Thunbergia laurifolia (TL) is plant widely distributed in South East Asia. TL leaves have been used as herbal infusion tea for treating fever and chemical intoxication. In this study, we examined the content of bioactive phenolic acids and flavonoids from leaves of TL in different solvent extraction. Besides, in vitro callus culture of TL was established. The rat skeletal muscle (L6) cell model was performed to investigate the glucose uptake stimulation. TL leaves extract with absolute methanol produced higher glucose uptake stimulation than those of the other solvent extracts. Interestingly, callus of TL was successful in producing high rosmarinic acid content over than intact leaves (increase up to 96-fold). The enriched rosmarinic acid callus extract significantly enhanced glucose uptake activity and elevated AMPK-α2 and GLUT4 mRNA expression. The established TL callus culture can be used as a promising source for production of biologically active phenolic acid with in vitro antidiabetic effect. Practical applications The boiled water extraction via hot infusion as herbal drinking tea could extract bioactive constituents, caffeic acid, rutin, and apigenin from Thunbergia laurifolia (TL) leaves, which enhanced glucose uptake activity. Moreover, the enriched rosmarinic acid in TL callus culture can be applied for promoting as a new source of rosmarinic acid, which exhibited antidiabetic activity.

Antioxidative effects of a novel dietary supplement Neumentix in a mouse stroke model

BackgroundDuring an acute stroke, reactive oxygen species are overproduced and the endogenous antioxidative defense systems are disrupted. Therefore, antioxidative therapy can be a promising scheme to reduce the severity of stroke. Neumentix is a novel antioxidative supplement produced from a patented mint line and contains a high content of rosmarinic acid (RA). Although Neumentix has proven diverse efficacy and safety in clinical trials, its effect on strokes is unclear. MethodsMice that were treated with Neumentix or vehicle for 14 days underwent transient middle cerebral artery occlusion (tMCAO) for 60 min. Mice were sacrificed 5 days after tMCAO. ResultsNeumentix preserved body weight after tMCAO, showed a high antioxidative effect in serum, and reduced infarction volume compared to the vehicle. The expression of 4-hydroxy-2-nonenal, Nε-(carboxymethyl) lysine, and 8-hydroxy-2′-deoxyguanosine was reduced in Neumentix-treated mice. ConclusionThe antioxidative effect of Neumentix was confirmed. This is the first report to demonstrate the antioxidative effect of Neumentix on strokes.