Alkanna Tinctoria Research Articles

Review on Bioactive Compound, Alkannin/Shikonin: Isolation, Biosynthesis, Formulations, Clinical trials, Patents and Toxicity with Phytochemicals and Pharmacological Properties of an Endangered Species, Alkanna tinctoria

Review on Bioactive Compound, Alkannin/Shikonin: Isolation, Biosynthesis, Formulations, Clinical trials, Patents and Toxicity with Phytochemicals and Pharmacological Properties of an Endangered Species, Alkanna tinctoria

Ratanjot (Alkanna tinctoria L.) Root Extract, Rich in Antioxidants, Exhibits Strong Antimicrobial Activity against Foodborne Pathogens and Is a Potential Food Preservative.

Natural and sustainable plant-based antioxidants and antimicrobials are highly desirable for improving food quality and safety. The present investigation assessed the antimicrobial and antioxidant properties of active components from Alkanna tinctoria L. (herb) roots, also known as Ratanjot root. Two methods were used to extract active components: microwave-assisted hot water (MAHW) and ethanolic extraction. MAHW extract yielded 6.29%, while the ethanol extract yielded 18.27%, suggesting superior Ratanjot root extract powder (RRP) solubility in ethanol over water. The ethanol extract showed significantly higher antioxidant activity than the MAHW extract. Gas Chromatography-Mass Spectrometry analysis revealed three major phenolic compounds: butanoic acid, 3-hydroxy-3-methyl-; arnebin 7, and diisooctyl pthalate. The color attributes (L*, a*, b*, H°ab, C*ab) for the ethanolic and MAHW extracts revealed significant differences (p < 0.05) in all the above parameters for both types of extracts, except for yellowness (b*) and chroma (C*ab) values. The ethanol extract exhibited antimicrobial activity against 14 foodborne bacteria, with a significantly higher inhibitory effect against Gram-positive bacteria (Listeria monocytogenes and Staphylococcus aureus) than the Gram-negative bacteria (Salmonella enterica serovar Typhimurium and Escherichia coli). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were both 25 mg/mL for the Gram-negative bacteria, while the MIC and MBC concentrations varied for Gram-positive bacteria (0.049-0.098 mg/mL and 0.098-0.195 mg/mL) and the antimicrobial effect was bactericidal. The antimicrobial activities of RRP extract remained stable under broad temperature (37-100 °C) and pH (2-6) conditions, as well as during refrigerated storage for 30 days. Application of RRP at 1% (10 mg/g) and 2.5% (25 mg/g) levels in a cooked chicken meatball model system prevented lipid oxidation and improved sensory attributes and retarded microbial growth during refrigerated (4 °C) storage for 20 days. Furthermore, the RRP extract was non-toxic when tested with sheep erythrocytes and did not inhibit the growth of probiotics, Lacticaseibacillus casei, and Lactiplantibacillus plantarum. In conclusion, the study suggests that RRP possesses excellent antimicrobial and antioxidant activities, thus making it suitable for food preservation.

Unveiling the photostability of the molecule of colour shikonin: From solution to the solid state

Shikonin, Shk, is a molecule of colour characterized by its natural phenolic-based compound composition, belonging to the naphthoquinone family. It is known for its low solubility in water and limited resistance to lightfastness. Shk is present in variable amount in the dried roots of Lithospermum erythrorhizon, Lithospermum officinale, and Alkanna tinctoria. The dyestuff extracted from these plants has historically been employed in Asia and Europe for textile dyeing, producing various shades of purple. Despite the historical significance of this dye, it has received limited attention in the context of cultural heritage studies. In the present work, Shk and model hydroxynaphthoquinone dyes were irradiated with polychromatic and monochromatic light in solution and in the solid state, mimicking paint mock-ups, to evaluate their photochemical stability. Low photoreaction quantum yield values (ϕR < 10−5) were observed in solution, providing quantitative evidence for the high stability of this molecule to light exposure. Chromatographic-mass spectrometric analyses carried out on the irradiated solutions and extracts from artificially aged Shk paint mock-ups, detected 5,8-dihydroxy-2-(1‑hydroxy-3-oxo-4-methyl-4-pentenyl)-l,4-naphthoquinone as degradation product, together with newly identified intermediate species involved in the complex photodegradation process. These indicate that, both in solution and in the solid state, the photoreaction of Shk proceeds with a similar mechanism. In the context of heritage science, this work contributes to highlight the potentialities of the adopted noninvasive approach, corroborated by results collected by chromatographic-mass spectrometric techniques, in identifying Shk in historical and archaeological specimens through its absorption and emission features and/or by detecting its degradation products which are, for the first time, characterized in terms of their UV–Vis absorption and emission spectral properties.

Co-Delivery of Dragon's Blood and Alkanna tinctoria Extracts Using Electrospun Nanofibers: In Vitro and In Vivo Wound Healing Evaluation in Diabetic Rat Model.

The increasing prevalence of diabetic wounds presents a significant challenge due to the difficulty of natural healing and various obstacles. Dragon's blood (DB) and Alkanna tinctoria (AT) are well recognized for their potent healing abilities, which include potent antibacterial and anti-inflammatory activities. In this study, electrospun nanofibers (NFs) based on polyvinyl pyrrolidone (PVP) were co-loaded with both DB and AT, aiming to magnify their efficacy as wound-dressing applications for diabetic wound healing. The evaluation of these NFs as wound dressings was conducted using a streptozotocin-induced diabetic rat model. Electrospun NFs were prepared using the electrospinning of the PVP polymer, resulting in nanofibers with consistent, smooth surfaces. The loading capacity (LC) of AT and DB into NFs was 64.1 and 70.4 µg/mg, respectively, while in the co-loaded NFs, LC was 49.6 for AT and 57.2 µg/mg for DB. In addition, X-ray diffraction (XRD) revealed that DB and AT were amorphously dispersed within the NFs. The loaded NFs showed a dissolution time of 30 s in PBS (pH 7.4), which facilitated the release of AT and DB (25-38% after 10 min), followed by a complete release achieved after 180 min. The antibacterial evaluation demonstrated that the DB-AT mixture had potent activity against Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus). Along with that, the DB-AT NFs showed effective growth inhibition for both P. aeruginosa and S. aureus compared to the control NFs. Moreover, wound healing was evaluated in vivo in diabetic Wistar rats over 14 days. The results revealed that the DB-AT NFs improved wound healing within 14 days significantly compared to the other groups. These results highlight the potential application of the developed DB-AT NFs in wound healing management, particularly in diabetic wounds.

Chitosan/bentonite composite as a green mordant for Alkanna tinctoria root dyeing of wool fabrics

Chitosan/bentonite (CS/BT) composite material was used to pretreat wool fabric to optimize its dyeing performance. CS/BT composite was prepared and XRD patterns showed that chitosan intercalated between bentonite layers. The wool was pretreated with chitosan (CS) and CS/BT before dyeing with Alkanna tinctoria (AT) root. FTIR and SEM indicated that CS/BT can increase the positive surface charge of wool and be more evenly and densely distributed on wool fibers than CS. After treating wool with CS/BT, exhaustion rate of AT root increased from 40% to 80%, delta E value decreased from 3.79 NBS to 0.62 NBS. At the same time, there is no loss of color fastness and tensile strength while CS treated wool has varying degrees of loss in both aspects. In summary, BT/CS can be used as a green mordant for AT root dyeing wool to improve dyeing performance.

Effects of fungal endophytes and arbuscular mycorrhizal fungi on growth of Echium vulgare and alkannin/shikonin and their derivatives production in roots

Endophytic fungi as well as arbuscular mycorrhizal fungi (AMF) are known to stimulate plant growth and production of secondary metabolites in medicinal plants. Here, 10 endophytic fungi isolated from roots of wild Alkanna tinctoria plants and 5 AMF purchased from the Glomeromycota in vitro collection were evaluated, during two successive three-month greenhouse experiments, on the growth of Echium vulgare and alkannin/shikonin and their derivatives (A/Sd) production in the roots. Some of the endophytic fungi tested significantly increased plant growth parameters as compared to the control: Cladosporium allicinum, Cadophora sp., Clonostachys sp., Trichoderma hispanicum and Leptosphaeria ladina increased root volume, Plectosphaerella sp. And T. hispanicum root fresh weight and root water retention and T. hispanicum plant water retention. However, none of these fungi impacted A/Sd production. Conversely, none of the AMF strains tested impacted plant growth parameters, but those inoculated with Rhizophagus intraradices MUCL 49410 had a significantly higher concentration of alkannin/shikonin (A/S), acetyl-A/S, β,β– dimethylacryl-A/S, isovaleryl-A/S and total A/Sd, compared to the control plants. Further studies are needed to investigate the mechanisms involved in the production of A/Sd in plants associated to specific endophytic fungi/AMF and on the cultivation conditions required for optimal production of these compounds.

Using extract from alkanet (Alkanna tinctoria) as a source of both a red lipid stain and a blue counterstain for histology

ABSTRACT Alkanet (Alkanna tinctoria) is a plant native to and cultivated in parts of Europe, Asia and the Middle East. It has been used for thousands of years as a medicinal agent and as a colorant for textiles, food and cosmetics. An extract from the root of this plant has been used with a mordant to stain nuclei. We describe here the versatility of different extracts from this plant to stain lipids red and to counterstain certain other tissue elements blue. The color variation and selective differential staining is due to solvent polarity and pH. Extracts contain numerous chemical species, all of which are derivatives of the indicator dye, naphthazurin. Our red extract is nonionic below pH 7 and partitions from its somewhat polar solvent of 100% isopropanol to nonpolar lipids. The blue extract is dianionic at high pH in 70% isopropanol and binds ionically to cationic tissue structures such as collagen, muscle and cytoplasm of other cells.

Arbuscular mycorrhizal fungi impact the production of alkannin/shikonin and their derivatives in Alkanna tinctoria Tausch. grown in semi-hydroponic and pot cultivation systems.

Alkanna tinctoria Tausch. is a medicinal plant well-known to produce important therapeutic compounds, such as alkannin/shikonin and their derivatives (A/Sd). It associates with arbuscular mycorrhizal fungi (AMF), which are known, amongst others beneficial effects, to modulate the plant secondary metabolites (SMs) biosynthesis. However, to the best of our knowledge, no study on the effects of AMF strains on the growth and production of A/Sd in A. tinctoria has been reported in the literature. Here, three experiments were conducted. In Experiment 1, plants were associated with the GINCO strain Rhizophagus irregularis MUCL 41833 and, in Experiment 2, with two strains of GINCO (R. irregularis MUCL 41833 and Rhizophagus aggregatus MUCL 49408) and two native strains isolated from wild growing A. tinctoria (R. irregularis and Septoglomus viscosum) and were grown in a semi-hydroponic (S-H) cultivation system. Plants were harvested after 9 and 37 days in Experiment 1 and 9 days in Experiment 2. In Experiment 3, plants were associated with the two native AMF strains and with R. irregularis MUCL 41833 and were grown for 85 days in pots under greenhouse conditions. Quantification and identification of A/Sd were performed by HPLC-PDA and by HPLC-HRMS/MS, respectively. LePGT1, LePGT2, and GHQH genes involved in the A/Sd biosynthesis were analyzed through RT-qPCR. In Experiment 1, no significant differences were noticed in the production of A/Sd. Conversely, in Experiments 2 and 3, plants associated with the native AMF R. irregularis had the highest content of total A/Sd expressed as shikonin equivalent. In Experiment 1, a significantly higher relative expression of both LePGT1 and LePGT2 was observed in plants inoculated with R. irregularis MUCL 41833 compared with control plants after 37 days in the S-H cultivation system. Similarly, a significantly higher relative expression of LePGT2 in plants inoculated with R. irregularis MUCL 41833 was noticed after 9 versus 37 days in the S-H cultivation system. In Experiment 2, a significant lower relative expression of LePGT2 was observed in native AMF R. irregularis inoculated plants compared to the control. Overall, our study showed that the native R. irregularis strain increased A/Sd production in A. tinctoria regardless of the growing system used, further suggesting that the inoculation of native/best performing AMF is a promising method to improve the production of important SMs.

Bacterial responses to plant antimicrobials: the case of alkannin and shikonin derivatives.

Alkannin, shikonin and their derivatives (A/S) are secondary metabolites produced in the roots of certain plants of the Boraginaceae family such as Lithospermum erythrorhizon Siebold & Zucc. and Alkanna tinctoria (L.) Tausch. These naphthoquinones express anti-cancer, wound healing, and antimicrobial activities. To study the interactions between endophytic bacteria isolated from A. tinctoria and the antimicrobials A/S, endophytic bacteria known to be resistant to the compounds were screened for their effect on A/S in liquid medium. Thereafter, the strain Pseudomonas sp. R-72008, was selected and tested for its ability to modify A/S in nutrient medium and minimal medium with A/S as sole carbon source. Bacterial growth was recorded, and high performance liquid chromatography-diode array and ultra-high performance liquid chromatography-electrospray ionization-mass spectrometry analyses were performed to detect and quantify metabolites. In nutrient medium inoculated with R-72008, a decrease in the amount of A/S monomers initially present was observed and correlated with an increase of A/S oligomers. Moreover, a significant decrease of initial A/S monomers in minimal medium was correlated with bacterial growth, showing for the first time that a bacterial strain, Pseudomonas sp. R-72008, was able to use the naphthoquinones A/S as sole carbon source. This study opens new perspectives on the interactions between bacteria and plant antimicrobials.

Harnessing Natural Colorant From Brown Algae TURBINARIA CONOIDES ( J. AGARDH) KUETZ For Fabric Dyeing: A Sustainable Eco- Friendly Approach

The demand for natural dyes in the modern world is increasing day by day. These resources have become important for their use in food, pharmaceuticals and textiles Industries, instead of their synthetic dyes. Using these resources we can protect human health and prolong life on the earth. This research focuses on Turbinaria conoides, a brown seaweed collected from Olakuda, Ramanathapuram coast, Tamil Nadu, India (9.2876° N, 79.3129° E). A crude dye was extracted from the seaweed and used to treat cotton fabric using the pre-mordanting technique. To achieve different shades of colors, Turbinaria conoides was combined with other dye-yielding plant materials, including Vachellia nilotica, Oldenlandia umbellate, Rubia cordifolia, and Alkanna tinctoria. The phytochemical analysis confirmed the presence of bioactive compounds in Turbinaria conoides. Colorfastness tests were conducted to evaluate the dye's stability on the fabric. The study concludes that natural dyes hold great potential and interest, especially Turbinaria conoides. India's vast plant resource base presents opportunities for further research and documentation of natural dye usage. Sustainable and eco-friendly practices in the textile industry can be fostered through algal-sourced pigments, contributing to environmental protection and an eco-aware society. Future aspects include a focus on environment-friendly dyes, affordable production methods, and exploring the full potential of natural dyestuffs for various applications, including waste utilization and job creation.

Uncovering Genomic Features and Biosynthetic Gene Clusters in Endophytic Bacteria from Roots of the Medicinal Plant Alkanna tinctoria Tausch as a Strategy To Identify Novel Biocontrol Bacteria.

The world's population is increasing at a rate not seen in the past. Agriculture, providing food for this increasing population, is reaching its boundaries of space and natural resources. In addition, changing legislation and increased ecological awareness are forcing agriculture to reduce its environmental impact. This entails the replacement of agrochemicals with nature-based solutions. In this regard, the search for effective biocontrol agents that protect crops from pathogens is in the spotlight. In this study, we have investigated the biocontrol activity of endophytic bacteria isolated from the medicinal plant Alkanna tinctoria Tausch. To do so, an extensive collection of bacterial strains was initially genome sequenced and in silico screened for features related to plant stimulation and biocontrol. Based on this information, a selection of bacteria was tested in vitro for antifungal activity using direct antagonism in a plate assay and in planta with a detached-leaf assay. Bacterial strains were tested individually and in combinations to assess the best-performing treatments. The results revealed that many bacteria could produce metabolites that efficiently inhibit the proliferation of several fungi, especially Fusarium graminearum. Among these, Pseudomonas sp. strain R-71838 showed a strong antifungal effect, in both dual-culture and in planta assays, making it the most promising candidate for biocontrol application. Using microbes from medicinal plants, this study highlights the opportunities of using genomic information to speed up the screening of a taxonomically diverse set of bacteria with biocontrol properties. IMPORTANCE Phytopathogenic fungi are a major threat to global food production. The most common management practice to prevent plant infections involves the intensive use of fungicides. However, with the growing awareness of the ecological and human impacts of chemicals, there is a need for alternative strategies, such as the use of bacterial biocontrol agents. Limitations in the design of bacterial biocontrol included the need for labor-intensive and time-consuming experiments to test a wide diversity of strains and the lack of reproducibility of their activity against pathogens. Here, we show that genomic information is an effective tool to select bacteria of interest quickly. Also, we highlight that the strain Pseudomonas sp. R-71838 produced a reproducible antifungal effect both in vitro and in planta. These findings build a foundation for designing a biocontrol strategy based on Pseudomonas sp. R-71838.

Effects of herbal mixture on burned rat skin infected with Pseudomonas aeruginosa

Purpose: The purpose of this study is to investigate the healing impact of an ointment derived from the Alkanna tinctoria plant upon Pseudomonas aeruginosa-induced infections. &#x0D; Material and Methods: In this study, 18 male adolescent rats (mean age 6 weeks) weighed an average of 180 g were used. Animals were divided into 3 groups. Group 1: Control group (consisting of burns, no treatment was done), Group 2 (P. aeruginosa): Burn was created and infected with P. aeruginosa, Group 3 (Cream): P. aeruginosa was used to infect the burns area and the herbal mixture was administered twice a day, once in the morning and once in the afternoon. Under anesthesia, the backs of the rats were shaved, and a specially produced steel bar with a diameter of 1*1 cm was immersed in boiling water for 15 seconds before being applied to their backs for 20 seconds. The burned area was subsequently infected with the ATCC Pseudomonas aeruginosa strain, and samples were collected 24 hours later. To detect bacterial growth in this area, the samples were inoculated on blood and EMB (Eosin Methylene Blue) media in a microbiology laboratory. After inoculation, the animals were placed in separate sterile cages and randomly divided into three groups. Once the growth was observed, the tissue and blood samples were harvested from the rats on the 2nd, 7th, 14th, and 21st days.&#x0D; Results: Epithelial regeneration in this group was more prominent. Vascularization was remarkable on the 2nd day, especially in the group in where we induced a burn and applied the ointment. VEGF levels increased more in the ointment group than in that of others. On the 2nd day of the study, the average bacterial count was 105 in sample of both 2nd and 3rd groups. At the end of the study, while the average of bacterial count was increased in the 2nd group, the average of bacterial count was decreased in the 3rd group.&#x0D; Conclusion: It was concluded that the ointment obtained from A. tinctorial successfully repaired the epithelial tissue and contributed to the healing of wounds by modifying increasing VEGF in the blood. However, further research is needed before this ointment can be highly recommended for therapeutic usage

Screening of medicinal plant extracts as novel DNA gyrase inhibitors

Bioactivities of a number of medicinal plants; Alkanna tinctoria (L.) Tausch, Alnus glutinosa (L.) Gaertn., Calamintha nepeta Willk. and C. nepeta, Centaurea iberica Trevir. ex Spreng., Citrus paradisi Macfad., C. paradisi, Citrus sinensis (L.) Osbeck, Colutea cilicica Boiss. & Balansa, Cotinus coggygria Scop., Cuscuta arvensis Beyr. ex Engelm., Equisetum palustre L., Lapsana communis L., Laurus nobilis L., Olea europea L., Plantago major L., Rhus coriaria L, Salvia verticillata L., Sambucus ebulus L., Sedum acre L, Thymus capitatus (L.) Hoffmanns. & Link, T. capitatus, Thymbra spicata L., T. spicata (n: 20), which are used for the prevention and treatment of diverse diseases, were investigated. The antimicrobial activities of extracts were evaluated using broth microdilution assay. The cytotoxicities of extracts were investigated on HeLa cell line by MTT assay. Statistical analysis was performed using GraphPad Prism (5.0). The effects of the extracts, which have the highest antimicrobial activity, on the Escherichia coli and Staphylococcus aureus DNA gyrase gene expression were determined by using quantitative reverse transcription polymerase chain reaction (qRT-PCR). The MICs (µg/ml) of extracts were determined as 32-64, 2-128, 8-128,1-128, 4-128 against Gram-positive, Gram-negative bacteria, yeasts, dermatophytes, and Mycobacterium spp., respectively. No cytotoxicity has been observed in plant extracts tested. DNA gyrase activity was determined for T. capitatus-SFE (128 µg/mL) and L. nobilis-Hx (128 µg/mL) extracts according to the inhibition of DNA gyrase gene expression. Overall, T. capitatus-SFE and L. nobilis-Hx are good candidates for further antimicrobial studies.

A Review on Henna

Plants produce henna. Medicine is made from the leaf. Contrast henna with henna root, also known as alkanna root (Alkanna tinctoria). Since ancient times, nature has been a rich supply of medicinal substances. Based on the traditional medical applications of these plants, a remarkable number of contemporary pharmaceuticals have been extracted from natural sources. A common example of such a plant is henna, also known as Persian henna or Lawsonia inermis, a bushy, flowering tree that can be found in Australia, Asia, and along the Mediterranean beaches of Africa.

The Potential of Medicinal Plants and Natural Products in the Treatment of Burns and Sunburn—A Review

Treating burns remains a challenge for modern medicine, especially in developing countries that cannot afford expensive, advanced therapies. This review article summarises clinical and animal model studies of botanical preparations and their mixtures in treating burn wounds and sunburn. Articles available in electronic databases such as PubMed, Scopus, Web of Science, Science Direct and Google Scholar, published in English in 2010-2022, were considered. In the described clinical trials, it was shown that some herbal preparations have better effectiveness in treating burn wounds, including shortening the healing time and reducing inflammation, than the conventional treatment used hitherto. These herbal preparations contained extracts from Albizia julibrissin, Alkanna tinctoria, Aloe vera, Arnebia euchroma, Betula pendula and Betula pubescens, Centella asiatica, Hippophaë rhamnoides, Juglans regia, Lawsonia inermis, and mixtures of Matricaria chamomilla and Rosa canina. Research on animal models shows that many extracts may potentially benefit the treatment of burn wounds and sunburn. Due to the diverse mechanism of action, antibacterial activity, the safety of use and cost-effectiveness, herbal preparations can compete with conventional treatment. The growing interest in alternative medicine and herbal medicine encourages further research. Not only single preparations but also their mixtures should be taken into account because the research conducted so far often suggests a synergistic effect of the ingredients.

Metabolic Profiles, Genetic Diversity, and Genome Size of Bulgarian Population of Alkanna tinctoria

Alkanna tinctoria (L.) Tausch Boraginaceae is a medicinal plant whose root is used for its antimicrobial and anti-inflammatory properties. A. tinctoria roots have been subject to numerous studies. However, the aerial parts have been explored less. The objective of the present study was to compare the chemical profile of aerial parts and roots as well as the total alkannin content in roots of 11 populations of the species from different floristic regions of Bulgaria. Methanolic extracts from 22 samples were analyzed by GC/MS. Phenolic, fatty, and organic acids, sterols, polyols, fatty alcohols, and sugars were identified. Ononitol (4-O-methyl-myo-inositol) was found as the main compound in the aerial parts. The total alkannin content in the roots was evaluated by the spectrophotometric method and compared with that of the commercial product. Populations with high alkannin content and rich in other bioactive compounds were identified. A relatively low genetic diversity in the studied populations was observed. The present study is the first comprehensive study on metabolite profiles and genetic diversity of the Bulgarian populations of A. tinctoria. The occurrence of ononitol in the aerial parts of the species is reported for the first time, as well as the phenolic acid profiles of the species in both aerial parts and roots. The results showed that aerial parts of the plant are also promising for use as a source of valuable biologically active substances.

Alkannin reverses lipopolysaccharides-induced inflammatory responses by suppressing mitogen-activated protein kinase and nuclear factor kappa-B signalling

ABSTRACT Rheumatoid arthritis (RA) is an inflammatory disease that seriously affects human health worldwide. Meanwhile, inflammation in RAW264.7 cells could lead to the progression of RA. Alkannin (ALK) is derived from Alkanna tinctoria and is known to exert anti-tumor effects. However, the function of ALK in inflammation of RAW264.7 cells remains unclear. Thus, this research sought to investigate the detailed function of ALK in inflammatory responses of RAW264.7 cells. To induce an inflammatory response, RAW264.7 cells were exposed to lipopolysaccharide (LPS). MTT assay was applied to examine cell viability. Enzyme-linked immunosorbent assay (ELISA) was used to assess the levels of inflammatory cytokines. Furthermore, the mechanism underlying ALK function in inflammatory responses was investigated using RT-qPCR and western blotting. The data revealed that LPS significantly increased the expression of cyclooxygenase 2 (COX-2), Interleukin (IL)-1β, inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), and IL-6, whereas ALK reversed this effect. ALK also restored LPS-induced nuclear factor kappa-B (NF-κB) activation by inhibiting the downregulation of p-inhibitor kappa B alpha (IκBα). LPS elevated p-extracellular regulated protein kinases 1/2 (ERK1/2), phosphorylated p38 (p-p38), and phosphorylated -c-Jun N-terminal kinase (p-JNK) levels, which were markedly decreased in the presence of ALK. In summary, Alkannin attenuated LPS-induced inflammation by inhibiting NF-κB and MAPK signaling. Thus, our research might provide a new theoretical basis for exploring new strategies against RA.

Nanostructured lipid carriers of alkannins and shikonins: Experimental design, characterization and bioactivity studies

The chiral pair alkannin and shikonin and their ester derivatives (A/S), are lipophilic secondary metabolites possessing a wide spectrum of pharmacological activities. Their incorporation in lipid-based drug delivery systems can enhance their therapeutic efficacy, especially for dermal or transdermal applications. In this work, A/S in the form of oily extracts and isolated mixture of pigments from Alkanna tinctoria roots were entrapped in lipid based nanocarriers for dermal applications. In both cases, Experimental Design was carried out to evaluate the effect of process parameters on mean particle size, ζ-potential and entrapment efficiency, aiming to identify the optimum process conditions. The optimum oily extract loaded liposomes showed a mean particle size of 106 nm, ζ-potential value of −45.1 mV and entrapment efficiency (EE%) up to 72%. On the other hand, the optimum nanocarriers for the incorporation of A/S mixture showed a mean particle size of 138 nm, ζ-potential value of −15.6 mV and EE% up to 56%. The in vitro release profiles for both drug delivery systems were satisfactory and revealed that, in both cases, the release of the active pharmaceutical ingredients from the nanocarriers is controlled by diffusion. Moreover, we evaluated the effect of the optimum formulations on the viability of human dermal fibroblasts and we found that, A/S mixture loaded nanocarriers enhance cell viability. In order to assess the effect of A/S derivatives loaded nanocarriers on human fibroblasts we targeted genes related to the antioxidant cellular response, cell aging and extracellular matrix organization. The findings showed that nanocarriers induced the antioxidant response, presented a positive impact on cells’ longevity and promoted extracellular matrix organization. Based on these findings, formulated nanocarriers can be used as a potential drug delivery system for dermal applications in cosmetic products.

Environmental friendly bio-dyeing of silk using Alkanna tinctoria based Alkannin natural dye

Application of bio-dyes, especially plant-based, is becoming increasingly popular in many industries such as textiles, food and pharmaceuticals. This trend could be explained by the ecological and durable nature of these compounds. The current paper aimed to investigate the microwave-assisted isolation of Alkannin dye from Alkanna tinctoria followed by the dyeing of bio-mordanted silk. For the purpose, acid solubilized, water solubilized, and acid-methanol solubilized extracts were obtained. Both respective extracts and silk fabrics were microwave irradiated for 2, 4 and 6 min. To ensure a green and clean dyeing process, bio-mordants were used. It is observed that acid solubilized extract applied at 55 °C for 55 min having 7 g/100 mL of NaCl salt as the exhausting agent has given extraordinary higher color strength (K/S) onto Microwave treatment (MW) treated silk fabric. Colorfastness tests using ISO-recommended requirements found that bio-mordants outperformed chemical mordants in terms of color depth and fastness. According to this study, it was proven that MW improved the dyeing performance of Alkannin in an acid-solubilized medium, where as in addition to the application of bio-mordants in comparison with sustainable chemical mordants have improved color characteristics of silk fabric dyed with Alkanna tinctoria extract.

INVESTIGATING LOW DOSE OF WEED MEDIATED SILVER NANOPARTICLES AGAINST PATHOGENIC BACTERIA

Weeds are undesirable and unpleasant plants with robust growth. Around 18,000 weed species outcast native biodiversity and responsible for tremendous economic losses. Plants were used to treat numerous ailments since ancient times. Antimicrobial resistance (AMR) is an emerging global health problem, where approximately 700,000 people lose their life each year from AMR infections. Minimum inhibitory concentration of Biogenic silver nanoparticles against pathogenic bacterial strain has not been reported previously. This study aims to elucidate lowest biogenic Alkanna tinctoria mediated silver nanoparticles dose to treat bacterial infections. Qualitative phytochemical analysis performed using standard tests. The synthesized silver nanoparticles were characterized visually, UV–Vis spectroscopy and using X-ray diffraction. Minimum inhibitory concentration of biogenic silver nanoparticles was tested against gram positive (S. aureus) and gram-negative bacteria (E. coli). Alkanna tinctoria leave were positive for alkaloids, carbohydrate, saponins, amino acids, and fixed oil. The synthesized silver nanoparticles were in range of 60 to 73nm. Silver nanoparticles significantly affected E. coli as compared to S. aureus. The MIC for E. coli was recorded as 46.87 μg/mL and 93.75 μg/mL was effective against S.aureus. Search of naive plants as novel therapeutic agents along with nanotechnology can bring revolution in the field of drug development and medical sciences.