Facilitation of infectious and non-infectious wound healing using Morus nigra fruit extract ointment: An in vitro and in vivo study.

Accelerating wound healing, as well as preventing infection and scar formation are among the most important medical challenges. This study aims to examine the antimicrobial, immunomodulatory, and anticancer properties of Morus nigra. The antimicrobial activities of ripe and unripe M. nigra fruit (MNF) extracts were tested. HPLC was employed to measure the components in the extract. Oserin ointment was made with 8 % extract. To test the ointment, 48 Wistar rats were randomly assigned into eight groups. The ointment was used daily by treating the wounds. Tissue histology and wound healing were assessed over nine days. Comparative evaluation of wound healing was conducted by analyzing TGF-ß, TNF-α, and IL-1 mRNA levels. Finally, cytotoxic effects on AGS cancer and NIH-3 T3 fibroblast cells were examined. The ANOVA test and Prsim program were used for statistical analysis. Unripe MNF extract had good antimicrobial properties in standard and nosocomial strains. The most abundant compound in the extract was ascorbic acid (0.0441 mg/10 mg extract), followed by naringenin and gallic acid. In all groups treated with MNF extract ointment, a significant reduction in wound area was observed compared to other groups (p < 0.05). After six days of treatment, the microbial load was uncountable. In the microscopic studies of the wounds, a significant increase was observed in fibroblasts, angiogenesis, and in neutrophils in the first days as well as a decrease in the final days. The treatment caused a significant decline in the expression of IL-1 and TNF-α genes, as well as an increase in the expression of TGF-ß (p < 0.05). This extract had no significant cytotoxic effects on human fibroblast cells (p > 0.05). In general, it can be concluded that the unripe MNF extract ointment can be a suitable option for the treatment of infectious and non-infectious skin wounds.

Specific Antimicrobial Activities Revealed by Comparative Evaluation of Selected Gemmotherapy Extracts.

Nowadays, unprecedented health challenges are urging novel solutions to address antimicrobial resistance as multidrug-resistant strains of bacteria, yeasts and moulds are emerging. Such microorganisms can cause food and feed spoilage, food poisoning and even more severe diseases, resulting in human death. In order to overcome this phenomenon, it is essential to identify novel antimicrobials that are naturally occurring, biologically effective and increasingly safe for human use. The development of gemmotherapy extracts (GTEs) using plant parts such as buds and young shoots has emerged as a novel approach to treat/prevent human conditions due to their associated antidiabetic, anti-inflammatory and/or antimicrobial properties that all require careful evaluations. Seven GTEs obtained from plant species like the olive (Olea europaea L.), almond (Prunus amygdalus L.), black mulberry (Morus nigra L.), walnut (Juglans regia L.), blackberry (Rubus fruticosus L.), blackcurrant (Ribes nigrum L.) and bilberry (Vaccinium myrtillus L.) were tested for their antimicrobial efficiency via agar diffusion and microbroth dilution methods. The antimicrobial activity was assessed for eight bacterial (Bacillus cereus, Staphylococcus aureus, Salmonella enterica subsp. enterica, Proteus vulgaris, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa and Listeria monocytogenes), five moulds (Aspergillus flavus, Aspergillus niger, Aspergillus ochraceus, Penicillium citrinum, Penicillium expansum) and one yeast strain (Saccharomyces cerevisiae). The agar diffusion method revealed the blackberry GTE as the most effective since it inhibited the growth of three bacterial, four moulds and one yeast species, having considered the total number of affected microorganism species. Next to the blackberry, the olive GTE appeared to be the second most efficient, suppressing five bacterial strains but no moulds or yeasts. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were then determined for each GTE and the microorganisms tested. Noticeably, the olive GTE appeared to feature the strongest bacteriostatic and bactericidal outcome, displaying specificity for S. aureus, E. faecalis and L. monocytogenes. The other GTEs, such as blueberry, walnut, black mulberry and almond (the list indicates relative strength), were more effective at suppressing microbial growth than inducing microbial death. However, some species specificities were also evident, while the blackcurrant GTE had no significant antimicrobial activity. Having seen the antimicrobial properties of the analysed GTEs, especially the olive and black mulberry GTEs, these could be envisioned as potential antimicrobials that might enhance antibiotic therapies efficiency, while the blackberry GTE would act as an antifungal agent. Some of the GTE mixtures analysed have shown interesting antimicrobial synergies, and all the antimicrobial effects observed argue for extending these studies to include pathological microorganisms.

Herbal Technological Prospects of Morus nigra L.: A Systematic Patent Analysis Review.

BACKGROUND: Morus nigra L. is a plant with significant potential for drug development due to the presence of numerous bioactive compounds in its various parts. OBJECTIVES: This article aims to compile the technological perspectives of Morus nigra L. towards drug development and therapeutic indications based on registered patents in databases. METHODS: The study analyzed patents published within the last five years, focusing on products derived from different parts of the Morus nigra L. plant. Patent databases such as the European Patent Office (EPO), the United States Patent and Trademark Office (USPTO), the World Intellectual Property Organization (WIPO), and the National Institute of Industrial Property Databases (INPI) were examined. RESULTS: A total of 45 patents were categorized by country of origin, type of applicant, extraction method, and therapeutic indications. China had the highest number of patent filings (43.48%), and private companies were the primary technology patent holders (38.64%). Noteworthy extraction methods included ultrasound-assisted extraction, decoction, infusion, and maceration. The most utilized plant parts were leaves (44.44%), followed by fruits (35.56%), root bark (15.56%), and stems (4.44%). The main therapeutic indications identified were the treatment of hyperglycemia and dyslipidemia (43.33%), along with digestive problems, cosmetics, nutrition, and cleaning applications. CONCLUSION: The study of patents covers discoveries and advancements often absent in scientific articles, making a review focused on this advanced information crucial for expanding existing scientific knowledge. Even if some therapies have been explored previously, patents can reveal innovative approaches and fresh perspectives that contribute to sustained scientific progress.

Phytoconstituent Profiles Associated with Relevant Antioxidant Potential and Variable Nutritive Effects of the Olive, Sweet Almond, and Black Mulberry Gemmotherapy Extracts.

The extracts of whole plants or specific organs from different plant species are gaining increasing attention for their phytotherapy applications. Accordingly, we prepared standardized gemmotherapy extracts (GTEs) from young shoots/buds of olive (Olea europaea), sweet almond (Prunus amygdalus), and black mulberry (Morus nigra), and analyzed the corresponding phytonutrient profiles. We identified 42, 103, and 109 phytonutrients in the olive, almond, and black mulberry GTEs, respectively, containing amino acids, vitamins, polyphenols, flavonoids, coumarins, alkaloids, iridoids, carboxylic acids, lignans, terpenoids, and others. In order to assess the physiological effects generated by the GTEs, we developed a translational nutrition model based on Drosophila melanogaster and Cyprinus carpio. The results indicate that GTEs could influence, to a variable extent, viability and ATP synthesis, even though both are dependent on the specific carbohydrate load of the applied diet and the amino acid and polyphenol pools provided by the GTEs. It seems, therefore, likely that the complex chemical composition of the GTEs offers nutritional properties that cannot be separated from the health-promoting mechanisms that ultimately increase viability and survival. Such an approach sets the paves the way for the nutritional genomic descriptions regarding GTE-associated health-promoting effects.

Plant lectins as versatile tools to fight coronavirus outbreaks.

The S protein forming the homotrimeric spikes of pathogenic beta-coronaviruses, such as MERS-CoV, SARS-CoV and SARS-CoV-2, is a highly glycosylated protein containing mainly N-glycans of the complex and high-mannose type, as well as O-glycans. Similarly, the host cell receptors DPP4 for MERS-CoV and ACE2 for SARS-CoV and SARS-CoV-2, also represent N- and O-glycosylated proteins. All these glycoproteins share common glycosylation patterns, suggesting that plant lectins with different carbohydrate-binding specificities could be used as carbohydrate-binding agents for the spikes and their receptors, to combat COVID19 pandemics. The binding of plant lectins to the spikes and their receptors could mask the non-glycosylated receptor binding domain of the virus and the corresponding region of the receptor, thus preventing a proper interaction of the spike proteins with their receptors. In this review, we analyze (1) the ability of plant lectins to interact with the N- and O-glycans present on the spike proteins and their receptors, (2) the in vitro and in vivo anti-COVID19 activity already reported for plant lectins and, (3) the possible ways for delivery of lectins to block the spikes and/or their receptors.

The inhibitory effect of Cannabis Sativa L. and Morus nigra L. against lipid peroxidation in goat liver and brain homogenates / O efeito inibitório de Cannabis Sativa L. e Morus nigra L. contra a peroxidação lipídica em homogenatos de fígado e cérebro de cabra

The present study was aimed to evaluate the antioxidant potential and inhibitory effect of Cannabis sativa and Morus nigra against lipid peroxidation in goat brain and liver homogenates. The formation of free radicals, highly reactive oxygen species (ROS) and reactive nitrogen species (RNS) is a normal metabolic process for cellular signaling and countering the antigens. However, they may cause serious damage if they produced at amplified tolls. In addition, metabolic disorders also serve as sources of these reactive species. Although the issue can be addressed through supplements and other phytochemicals. In this study, two plant species were evaluated for their biological potential by employing a spectrum of antioxidant assays. The antioxidant activity was performed by lipid peroxidation assay. The water extract prepared from leaves of Cannabis sativa and Morus nigra showed significant (P<0.05) inhibition as compared to control i.e., 522.6±0.06 and 659.97±0.03 µg/mL against iron-induced lipid peroxidation in goat brain homogenate while the inhibitions were 273.54±0.04 and 309.18±0.05 µg/mL against nitroprusside induced lipid peroxidation of the brain. The iron and nitroprusside induced lipid peroxidation was also significantly inhibited by leaf extracts of Cannabis sativa and Morus nigra in liver homogenates such as 230.63±0.52 and 326.91±0.01 µg/mL (iron-induced) while 300.47±0.07 and 300.47±0.07 µg/mL (nitroprusside induced), respectively. The extracts of Cannabis sativa extract showed promising activity (96.04±0.060%) against DPPH radicals while Morus nigra showed a moderate activity (34.11±0.120%). The results suggest that different accessions of Cannabis sativa and Morus nigra are a potential source of antioxidants and have a therapeutic effect against disease induced by oxidative stress and hence can be used for novel drug discovery and development.

O presente estudo teve como objetivo avaliar o potencial antioxidante e o efeito inibitório de Cannabis sativa e Morus nigra contra a peroxidação lipídica em homogenatos de cérebro e fígado de cabras. A formação de radicais livres, espécies altamente reativas de oxigênio (ROS) e espécies reativas de nitrogênio (RNS), é um processo metabólico normal para sinalização celular e combate aos antígenos. No entanto, eles podem causar sérios danos se forem produzidos em portagens ampliadas. Além disso, distúrbios metabólicos também servem como fontes dessas espécies reativas, embora o problema possa ser resolvido por meio de suplementos e outros fitoquímicos. Neste estudo, duas espécies de plantas foram avaliadas quanto ao seu potencial biológico, empregando um espectro de ensaios antioxidantes. A atividade antioxidante foi realizada por ensaio de peroxidação lipídica. O extrato de água preparado a partir de folhas de Cannabis sativa e Morus nigra mostrou inibição significativa (P < 0,05) em comparação com o controle, ou seja, 522,6 ± 0,06 e 659,97 ± 0,03 µg / mL contra peroxidação lipídica induzida por ferro em homogenato de cérebro de cabra, enquanto as inibições foram 273,54 ± 0,04 e 309,18 ± 0,05 µg / mL contra a peroxidação lipídica do cérebro induzida por nitroprussiato. A peroxidação lipídica induzida por ferro e nitroprussiato também foi significativamente inibida por extratos de folhas de Cannabis sativa e Morus nigra em homogenatos de fígado, como 230,63 ± 0,52 e 326,91 ± 0,01 µg / mL (induzida por ferro), enquanto 300,47 ± 0,07 e 300,47 ± 0,07 µg / mL (induzida por nitroprussiato), respectivamente. Os extratos do extrato de Cannabis sativa apresentaram atividade promissora (96,04 ± 0,060%) contra os radicais DPPH enquanto Morus nigra apresentou atividade moderada (34,11 ± 0,120%). Os resultados sugerem que diferentes acessos de Cannabis sativa e Morus nigra são uma fonte potencial de antioxidantes e têm efeito terapêutico [...].

The inhibitory effect of Cannabis Sativa L. and Morus nigra L. against lipid peroxidation in goat liver and brain homogenates

Abstract The present study was aimed to evaluate the antioxidant potential and inhibitory effect ofCannabis sativa and Morus nigra against lipid peroxidation in goat brain and liver homogenates. The formation of free radicals, highly reactive oxygen species (ROS) and reactive nitrogen species (RNS) is a normal metabolic process for cellular signaling and countering the antigens. However, they may cause serious damage if they produced at amplified tolls. In addition, metabolic disorders also serve as sources of these reactive species. Although the issue can be addressed through supplements and other phytochemicals. In this study, two plant species were evaluated for their biological potential by employing a spectrum of antioxidant assays. The antioxidant activity was performed by lipid peroxidation assay. The water extract prepared from leaves of Cannabis sativa and Morus nigra showed significant (P 0.05) inhibition as compared to control i.e., 522.6±0.06 and 659.97±0.03 µg/mL against iron-induced lipid peroxidation in goat brain homogenate while the inhibitions were 273.54±0.04 and 309.18±0.05 µg/mL against nitroprusside induced lipid peroxidation of the brain. The iron and nitroprusside induced lipid peroxidation was also significantly inhibited by leaf extracts of Cannabis sativa and Morus nigra in liver homogenates such as 230.63±0.52 and 326.91±0.01 µg/mL (iron-induced) while 300.47±0.07 and 300.47±0.07 µg/mL (nitroprusside induced), respectively. The extracts of Cannabis sativa extract showed promising activity (96.04±0.060%) against DPPH radicals while Morus nigra showed a moderate activity (34.11±0.120%). The results suggest that different accessions ofCannabis sativa and Morus nigra are a potential source of antioxidants and have a therapeutic effect against disease induced by oxidative stress and hence can be used for novel drug discovery and development.

Resumo O presente estudo teve como objetivo avaliar o potencial antioxidante e o efeito inibitório de Cannabis sativa e Morus nigra contra a peroxidação lipídica em homogenatos de cérebro e fígado de cabras. A formação de radicais livres, espécies altamente reativas de oxigênio (ROS) e espécies reativas de nitrogênio (RNS), é um processo metabólico normal para sinalização celular e combate aos antígenos. No entanto, eles podem causar sérios danos se forem produzidos em portagens ampliadas. Além disso, distúrbios metabólicos também servem como fontes dessas espécies reativas, embora o problema possa ser resolvido por meio de suplementos e outros fitoquímicos. Neste estudo, duas espécies de plantas foram avaliadas quanto ao seu potencial biológico, empregando um espectro de ensaios antioxidantes. A atividade antioxidante foi realizada por ensaio de peroxidação lipídica. O extrato de água preparado a partir de folhas de Cannabis sativa e Morus nigra mostrou inibição significativa (P 0,05) em comparação com o controle, ou seja, 522,6 ± 0,06 e 659,97 ± 0,03 µg / mL contra peroxidação lipídica induzida por ferro em homogenato de cérebro de cabra, enquanto as inibições foram 273,54 ± 0,04 e 309,18 ± 0,05 µg / mL contra a peroxidação lipídica do cérebro induzida por nitroprussiato. A peroxidação lipídica induzida por ferro e nitroprussiato também foi significativamente inibida por extratos de folhas de Cannabis sativa e Morus nigra em homogenatos de fígado, como 230,63 ± 0,52 e 326,91 ± 0,01 µg / mL (induzida por ferro), enquanto 300,47 ± 0,07 e 300,47 ± 0,07 µg / mL (induzida por nitroprussiato), respectivamente. Os extratos do extrato de Cannabis sativa apresentaram atividade promissora (96,04 ± 0,060%) contra os radicais DPPH enquanto Morus nigra apresentou atividade moderada (34,11 ± 0,120%). Os resultados sugerem que diferentes acessos de Cannabis sativa e Morus nigra são uma fonte potencial de antioxidantes e têm efeito terapêutico contra doenças induzidas por estresse oxidativo e, portanto, podem ser usados para a descoberta e desenvolvimento de novos medicamentos.

The inhibitory effect of Cannabis Sativa L. and Morus nigra L. against lipid peroxidation in goat liver and brain homogenates / O efeito inibitório de Cannabis Sativa L. e Morus nigra L. contra a peroxidação lipídica em homogenatos de fígado e cérebro de cabra

Abstract The present study was aimed to evaluate the antioxidant potential and inhibitory effect ofCannabis sativa and Morus nigra against lipid peroxidation in goat brain and liver homogenates. The formation of free radicals, highly reactive oxygen species (ROS) and reactive nitrogen species (RNS) is a normal metabolic process for cellular signaling and countering the antigens. However, they may cause serious damage if they produced at amplified tolls. In addition, metabolic disorders also serve as sources of these reactive species. Although the issue can be addressed through supplements and other phytochemicals. In this study, two plant species were evaluated for their biological potential by employing a spectrum of antioxidant assays. The antioxidant activity was performed by lipid peroxidation assay. The water extract prepared from leaves of Cannabis sativa and Morus nigra showed significant (P<0.05) inhibition as compared to control i.e., 522.6±0.06 and 659.97±0.03 µg/mL against iron-induced lipid peroxidation in goat brain homogenate while the inhibitions were 273.54±0.04 and 309.18±0.05 µg/mL against nitroprusside induced lipid peroxidation of the brain. The iron and nitroprusside induced lipid peroxidation was also significantly inhibited by leaf extracts of Cannabis sativa and Morus nigra in liver homogenates such as 230.63±0.52 and 326.91±0.01 µg/mL (iron-induced) while 300.47±0.07 and 300.47±0.07 µg/mL (nitroprusside induced), respectively. The extracts of Cannabis sativa extract showed promising activity (96.04±0.060%) against DPPH radicals while Morus nigra showed a moderate activity (34.11±0.120%). The results suggest that different accessions ofCannabis sativa and Morus nigra are a potential source of antioxidants and have a therapeutic effect against disease induced by oxidative stress and hence can be used for novel drug discovery and development.

Resumo O presente estudo teve como objetivo avaliar o potencial antioxidante e o efeito inibitório de Cannabis sativa e Morus nigra contra a peroxidação lipídica em homogenatos de cérebro e fígado de cabras. A formação de radicais livres, espécies altamente reativas de oxigênio (ROS) e espécies reativas de nitrogênio (RNS), é um processo metabólico normal para sinalização celular e combate aos antígenos. No entanto, eles podem causar sérios danos se forem produzidos em portagens ampliadas. Além disso, distúrbios metabólicos também servem como fontes dessas espécies reativas, embora o problema possa ser resolvido por meio de suplementos e outros fitoquímicos. Neste estudo, duas espécies de plantas foram avaliadas quanto ao seu potencial biológico, empregando um espectro de ensaios antioxidantes. A atividade antioxidante foi realizada por ensaio de peroxidação lipídica. O extrato de água preparado a partir de folhas de Cannabis sativa e Morus nigra mostrou inibição significativa (P < 0,05) em comparação com o controle, ou seja, 522,6 ± 0,06 e 659,97 ± 0,03 µg / mL contra peroxidação lipídica induzida por ferro em homogenato de cérebro de cabra, enquanto as inibições foram 273,54 ± 0,04 e 309,18 ± 0,05 µg / mL contra a peroxidação lipídica do cérebro induzida por nitroprussiato. A peroxidação lipídica induzida por ferro e nitroprussiato também foi significativamente inibida por extratos de folhas de Cannabis sativa e Morus nigra em homogenatos de fígado, como 230,63 ± 0,52 e 326,91 ± 0,01 µg / mL (induzida por ferro), enquanto 300,47 ± 0,07 e 300,47 ± 0,07 µg / mL (induzida por nitroprussiato), respectivamente. Os extratos do extrato de Cannabis sativa apresentaram atividade promissora (96,04 ± 0,060%) contra os radicais DPPH enquanto Morus nigra apresentou atividade moderada (34,11 ± 0,120%). Os resultados sugerem que diferentes acessos de Cannabis sativa e Morus nigra são uma fonte potencial de antioxidantes e têm efeito terapêutico contra doenças induzidas por estresse oxidativo e, portanto, podem ser usados ​​para a descoberta e desenvolvimento de novos medicamentos.

Hydroethanolic Extract of Morus nigra L. Leaves: A Dual PPAR-α/γ Agonist with Anti-Inflammatory Properties in Lipopolysaccharide-Stimulated RAW 264.7

Inhibition of systemic inflammation has been a beneficial strategy in treating several non-communicable diseases, which represent one of the major causes of mortality in the world. The Peroxisome Proliferator-Activated Receptors (PPAR) are interesting pharmacological targets, since they can act both through the metabolic and anti-inflammatory pathways. Morus nigra L. has flavonoids in its chemical composition with recognized anti-oxidant activity and often associated with anti-inflammatory activity. Therefore, this study aimed to evaluate the hydroethanolic extract of M. nigra leaves' ability to activate PPAR and promote anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated murine macrophage cells. The leaf extract was prepared by cold maceration, and the chemical profile was obtained by HPLC-DAD. Activation of PPAR α and γ was evaluated by the luciferase reporter assay. The anti-inflammatory activity was assessed by measuring the reactive oxygen species (ROS), nitric oxide (NO), and Tumor Necrosis Factor-α (TNF-α) in RAW 264.7 cells after stimulation with LPS from Escherichia coli. The HPLC-DAD analysis identified two major compounds: rutin and isoquercitrin. The extract showed agonist activity for the two types of PPAR, α and γ, although its major compounds, rutin and isoquercitrin, did not significantly activate the receptors. In addition, the extract significantly reduced the production of ROS, NO, and TNF-α. Treatment with the specific PPAR-α antagonist, GW 6471, was able to partially block the anti-inflammatory effect caused by the extract.

Black mulberry (Morus nigra) fruit extract alleviated AD-Like symptoms induced by toxic Aß protein in transgenic Caenorhabditis elegans via insulin DAF-16 signaling pathway.

Alzheimer's disease (AD) is one of the most severe neurodegenerative disorders. Recently, there is no effective treatment drug for AD. Morus nigra (M. nigra) is a black mulberry and widely distributed fruit in the Moraceae family with various undiscovered biological activities. The study aimed to investigate the potential anti-AD effect of M. nigra. Mulberry fruit extract (MF) was obtained from M. nigra and treated up to 1.00 mg/mL on transgenic AD Caenorhabditis elegans (C. elegans) models. MF inhibited Amyloid-ß (Aß)-induced paralysis symptoms by about 55.65 %, reduced Aß accumulation more than 50 % via immunoblotting, and suppressed over-sensitivity to exogenous serotonin in C. elegans. Furthermore, MF decreased the Aß oligomeric depositions in worm CL2006. MF activated the DAF-16 nuclear translocation and its downstream SOD-3 and GST-4. AD is a major age-related disorder. Therefore, MF treated for an aging test and proved to be expanded the lifespan of the worms up to 34.7 %. Besides, we have evaluated the MF in vivo antioxidative properties, where MF reduced reactive oxygen species (ROS) generations in C. elegans and remitted the activation of HSP-16.2 induced by the oxidative action of Juglone. Gene knockout and extended the lifespan of AD worms. However, RNA interference (RNAi) successfully silenced the daf-16 on the Aß phenotypic paralysis proved by MF effect. Our results indicate that MF alleviates AD-Like symptoms by activating the DAF-16 insulin signal pathway in C. elegans. Therefore, this MF study may provide new insights for mulberry application in safe AD treatment and clinical study.

Ameliorating Effect of the Total Flavonoids of Morus nigra L. on Prediabetic Mice Based on Regulation of Inflammation and Insulin Sensitization.

Prediabetes is a critical stage characterized by insulin resistance. Morus nigra L., an edible plant, is widely used in food and nutritive supplements and exhibits various pharmacological activities; however, its therapeutic effects and mechanisms on prediabetes have rarely been reported. In this research, the major components of total flavonoids of M. nigra L. (TFM) were identified, and TFM treatment was found to reduce prediabetes progressing to type 2 diabetes mellitus (T2DM) from 93.75 to 18.75%. The microbiota and next-generation sequencing combined with western blotting in vivo and in vitro demonstrated that TFM and its components ameliorated insulin resistance mediated by the suppressor of cytokine signaling and protein tyrosine phosphatase 1B, which benefited by maintaining intestinal homeostasis and restraining plasma levels of inflammatory factors. This study confirmed the T2DM prevention effect of TFM and revealed the underlying mechanism, setting the stage for the design of functional foods for diabetes prevention.

Prenylated flavonoids from Morus nigra and their insulin sensitizing activity.

Six undescribed prenylated flavonoids, nigragenons H-M, and four known compounds, were isolated from Morus nigra L. Their structures were elucidated through extensive analysis of spectroscopic data, and their absolute configurations were established by time-dependent density functional theory electronic circular dichroism (TDDFT ECD) calculation. The insulin sensitizing activities of all compounds were investigated using insulin-resistant 3T3-L1 adipocytes. At a high concentration (30 µM), all compounds except nigragenon I enhanced insulin-stimulated glucose uptake in insulin-resistant 3T3-L1 adipocytes. Furthermore, nigragenons J-L and the promoted adiponectin secretion in the model cells. Among them, nigragenon L showed the most potent effect at a low concentration of 10 µM, which was comparable to that of rosiglitazone at a concentration of 1 µM. Furthermore, using the Lantha Screen™ TR-FRET assay, nigragenon L was confirmed to be the ligand of PPARγ, showing potent binding affinity toward PPARγ with an IC50 value of 2.8 µM.

Phytochemical characterization of Morus nigra fruit ultrasound-assisted ethanolic extract for its cardioprotective potential.

The current work investigated the phytochemical profile of ultrasound-assisted ethanolic extract of Morus nigra (M. nigra) fruit. FTIR analysis of M. nigra fruit extract revealed the presence of alcohols (O-H), alkanes (C-H stretch), alkenes (C=C), and alkynes (C≡C). The HPLC analysis quantified the quercetin, gallic acid, vanillic acid, chlorogenic acid, syringic acid, cinnamic acid, sinapic acid, and kaempferol. Furthermore, the cardioprotective activity of ethanolic extract of M. nigra fruit was investigated. Cholesterol supplementation (2%) in the daily diet and exposure to cigarette smoke (2 cigarettes twice a day) were to induce hypertension in rats. The experimental animals were categorized into four groups: G0 (negative control), G1 (positive control), G2 (standard drug), and G3 (M. nigra fruit). The fruit extract administration at 300 mg/kg BW/day orally for 2 months significantly (p < .001) enhanced the activities of serum and cardiac tissue antioxidants in hypertensive rats. Meanwhile, the fruit extract reduced the elevated serum lipid profile while significantly increasing the high-density lipoproteins in G3 than G1 and G2. The increase in blood pressure, liver transaminases, and serum lactate dehydrogenase also reduced significantly in M. nigra fruit extract-treated rats. Histopathological findings revealed mild normalization of cardiac myocytes with central nuclei, branching, and cross-striations. Consequently, the M. nigra fruit extract exerted the cardioprotective potential via increasing the antioxidant enzymes and reducing the lipids, lactate dehydrogenase, liver transaminases, and blood pressure. The therapeutic potential of M. nigra fruit can be due to flavonols and phenolic acids. PRACTICAL APPLICATIONS: The present work quantified the Morus nigra fruit phytochemicals and its significant role in reducing lipid markers and blood pressure and improving antioxidant status in rats fed a hypercholesterolemic diet and exposed to cigarette smoke. Conclusively, the inclusion of M. nigra fruit in daily diet could improve the cardiac health of the individuals. Furthermore, the therapeutic potential of M. nigra fruit and its isolated constituents in modulating the gene expression against cardiac problems can explore after clinical trials and standardization in higher animals.

Optimization of Solvent-Free Microwave-Assisted Hydrodiffusion and Gravity Extraction of Morus nigra L. Fruits Maximizing Polyphenols, Sugar Content, and Biological Activities Using Central Composite Design.

Black mulberry, Morus nigra L. (family: Moraceae), is a healthy food and medicinal plant. Microwave hydrodiffusion and gravity (MHG) is one of the most innovative applications of solvent-free microwave extraction. The aim of this study was to optimize for the first time the MHG solvent-free extraction of polyphenols and sugars from M. nigra fruits. Optimization was carried out using a central composite design (CCD) with selected responses such as extraction yield, total polyphenol (TPC), flavonoid (TFC), anthocyanin (TAC), and sugar (TSC) contents, in addition to DPPH radical scavenging, and α-glucosidase (AGHi), lipase (Li), and xanthine oxidase (XOi) inhibition as tools to evaluate the best parameters for efficient and rapid extraction of black mulberry. The optimized extract was characterized in terms of the aforementioned parameters to validate the models, and was further analyzed for 36 individual polyphenols using HPLC-MS/MS. The optimized MHG extract was finally compared with traditional extracts, and demonstrated much better performance in terms of TPC, TAC, and Li, while the traditional extracts showed better XOi and AGHi. In conclusion, MHG is a valuable green technique for the production of non-degraded black mulberry polyphenol-rich extract and we suggest its larger use in the pharmaceutical and food industries.

Hydro-alcoholic extract of Morus nigra reduces fasting blood glucose and HbA1c% in diabetic patients, probably via competitive and allosteric interaction with alpha-glucosidase enzyme; a clinical trial and in silico analysis.

OBJECTIVES: 1-Deoxynojirimycin (1-DNJ), the main active component found in Morus nigra (black mulberry) is reported to be effective in controlling diabetes. We have evaluated the effect of hydro-alcoholic extract of M. nigra leaves on the fasting blood glucose (FBS) and hemoglobin A1c% (HbA1c%) in diabetic patients. Furthermore, we compared the interaction of 1-DNJ and glucose molecules with the alpha-glucosidase enzyme, which has a critical role in the lysis of glucose-based polymers in human cells. METHODS: 4% hydro-alcoholic extract was prepared from black mulberry leaves. Patients in treatment (n=50) and control (n=50) groups received 3 mL extract or placebo in water, respectively, and three times a day. Fasting blood glucose and HbA1c% were evaluated before and after three months of evaluation. Potential binding sites of 1-DNJ or glucose on the enzyme glucosidase found by docking study. Docking scores were obtained using an energy minimization method by Molegro Virtual Docker software. The Mean ± SD of each variable was compared between groups at the 95% significant level. RESULTS: Age mean ± SD was equal to 54.79 ± 9.203 (38-69) years. There was no significant difference between intervention and placebo groups considering FBS (p=0.633) but was for HbA1c% (p=0.0011), before treatment. After three months, both FBS and HbA1c% were significantly reduced in patients under mulberry leaves extract-treatment. FBS changed was from 182.23 ± 38.65 to 161.23 ± 22.14 mg/dL in treatment group (p<0.001) and from 178.45 ± 39.46 to 166.23 ± 29.64 mg/dL in control group (p<0.001). HbA1c was changed from 7.23 ± 0.25 to 6.13 ± 0.61% in treatment group (p<0.001) and from 7.65 ± 0.85 to 7.12 ± 0.33% in control group (p=0.854). Docking results showed that 1-DNJ binds more efficiently, and with a significant score than glucose, to human alpha-glucosidase. CONCLUSIONS: This clinical trial and virtual analysis showed that a hydro-alcoholic extract of black mulberry (M. nigra) leaf may be efficient in reducing the blood glucose and HbA1c% in diabetic patients. Furthermore, docking studies propose a competitive and allosteric regulation for herbal ingredients. Drug-development could be based on the presented idea in this report.

Antimicrobial and antioxidant activities of secondary metabolites from endophytic fungus Botryosphaeria fabicerciana (MGN23-3) associated to Morus nigra L.

This study was to evaluate the biological activity of the extract of Botryosphaeria fabicerciana isolated from leaves of Morus nigra. The volatile compounds from the crude extract were analysed by GC-MS which demonstrate that mellein and ß-orcinaldehyde were are the major compounds. The best minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extract was observed against Gram-positive bacteria, with a MIC of 15.6 µg/mL towards B. cereus and MIC of 62.5 µg/mL towards S. aureus and B. subtilis. MBC values of 31.25 µg/mL, 62.5 µg/mL, and 250 µg/mL were observed towards B. cereus, B. subtilis, and S. aureus, respectively. The cytotoxicity analyses showed CC50 of 115 µg/mL. The crude extract showed antioxidant activity by the DPPH, ABTS, and FRAP assays. Therefore, the extract of the endophytic fungus presented biotechnological potential as an antibacterial and antioxidant agent.

Chemical Components of the Fruits of Morus nigra Linn.: Methyl Caffeate as a Potential Anticancer Agent by Targeting 3-Phosphoglycerate Dehydrogenase.

Two previously undescribed compounds, moranigrine A (1) and morusamine (2), along with 18 known compounds were isolated from the fruits of Morus nigra Linn. and structurally characterized using spectroscopic data and electronic circular dichroism analyses. All isolates were evaluated for their inhibitory effects on the 3-phosphoglycerate dehydrogenase (PHGDH) enzyme, which catalyzes the first committed step for the synthesis of glucose-derived serine and is associated with many kinds of cancers. Among these compounds, methyl caffeate (3) exhibited effective inhibition against PHGDH and was directly bound to PHGDH based on the microscale thermophoresis method and the cellular thermal shift assay. Further biochemical assays revealed that 3 was a noncompetitive inhibitor with respect to the substrate of 3-phosphoglycerate and exhibited a concentration-dependent inhibition. Molecular docking demonstrated that 3 coordinated in an allosteric site of PHGDH with low binding energy. Meanwhile, 3 was selectively toxic to high PHGDH-expressing cancer cell lines and could cause apoptosis of cervical cancer cells in micromolar concentrations and could obviously inhibit tumor growth in the HeLa xenograft mouse model with low toxicities. Therefore, 3 could be developed as a potential inhibitor of PHGDH for the treatment of cancers. Our present study provides information about M. nigra as a functional food or pharmaceutical supplement in the application of cancer prevention and treatment.

Anthocyanins from Rubus fruticosus L. and Morus nigra L. Applied as Food Colorants: A Natural Alternative.

Given the importance of colour in the general acceptance or rejection of a product, the use of colorants is a widespread practice, particularly in the food industry. At the same time, with the increasing consumers' awareness of the health effects that some artificial colorants can exert, there is a growing tendency to prioritize foodstuffs containing natural additives. In this work, Morus nigra L. and Rubus fruticosus L. fruit juices were characterized in terms of anthocyanins, organic acids, free sugars, and tocopherols, as also regarding their bioactive properties. Given their richness in anthocyanins, this study also aimed to prepare different solid colouring formulations by the spray-drying technique, using as stabilizers maltodextrin and arabic gum. Six free sugars and two organic acids were detected in the fruit juices, as well as the four tocopherol isoforms. Two cyanidin derivatives were found in M. nigra (cyanidin-3-O-glucoside and cyanidin-O-rhamnoside) and other four in R. fruticosus (cyanidin-O-hexoside, cyanidin-3-O-glucoside, cyanidin-O-pentoside, and cyanidin-3-O-dioxaloilglucoside). The developed colouring formulations revealed a good stability over time, in terms of anthocyanin concentration and colour parameters, and revealed to be safe for consumption, either concerning their low microbial load and lack of cytotoxicity. Thus, they represent a promising natural alternative to the massively used artificial colorants.

Protective Effects of Morus nigra and Its Phytochemicals against Hepatotoxicity: A Review of Preclinical Studies.

BACKGROUND: Our liver has a variety of vital functions including removing poisons, storing energy, immunological roles, and secretory and excretory functions. It may face some kinds of diseases caused by viruses, hepatotoxic chemicals, drugs, alcohol, and inherited disorders. Oxidative stress and inflammation are in the core of mechanisms of liver damages induced by viruses or chemical agents. SUMMARY: Morus nigra (M. nigra), generally known as black mulberry, exhibited wide-spectrum pharmacological effects including antidiabetic, antinociceptive, anticancer, and hepatoprotective activities. Different parts of this plant particularly the fruit and leaf have shown beneficial effects on hepatocytes in cell culture and animal models of liver damages induced by chemicals (e.g., CCl4), drugs (e.g., paracetamol), diet (e.g., high fat), diabetes, etc. The beneficial effects of M. nigra on the liver are attributed to the presence of considerable amounts of phenolic compounds such as anthocyanins, flavonols, and phenolic acids. The present review is aimed to focus on the hepatoprotective activities of M. nigra and its phytochemicals and the mechanisms responsible for these activities. Key Messages: The evidence reviewed in this study can help design clinical trials on M. nigra in patients with liver disorders and develop a hepatoprotective herbal medicine.

Molecular and biological characterization of a new mulberry idaeovirus.

A virus-like disease with symptoms including mosaic structure, deformation, vein clearing and necrosis on the leaves and deformation, crumbling, and scab on the fruits was detected in black mulberry trees (Morus nigra L.) in Kayseri province of Turkey. A novel positive single-stranded RNA virus with a bipartite genome and the mulberry badnavirus 1 (MBV-1) were detected in the black mulberry trees by high throughput sequencing and bioinformatic analyses. The novel virus RNA1 (5,796/7 nt) encodes a polyprotein (1,808 aa, 204.31 kDa) with three conserved domains, [MTR (aa 294-705), Hel (aa 971-1,226) and RdRp (aa 1,348-1,788)], whereas RNA2 (2,243 nt) encodes two putative proteins, MP (374 aa, 40.98 kDa), and CP (272 aa, 30.59 kDa), separated by an intergenic region of 97 nt. The highest amino acids identities were 70, 57 and 70 % with raspberry bushy dwarf virus (RBDV) for ORF1, MP and CP genes, respectively. The genome organization and phylogenetic analyses suggested that the novel virus is likely a putative new member of the genus Idaeovirus and it has been tentatively named black mulberry idaeovirus (BMIV). Virus survey showed both the BMIV and MBV-1 are likely prevalent in the region. Seven complete (six Turkish and one Iranian) and 41 partial genome sequences of the BMIV isolates revealed moderate genetic diversity (0.033 ± 0.001 %, 0.020 ± 0.002 % and 0.016 ± 0.002 % for RNA1, RNA2, and partial genomes, respectively). Both the BMIV and MBV-1 were detected in all tested pollens (n = 24, 100 %), in seed-borne balck mulberry saplings (n = 96, 100 %).This situation clearly revealed the potential spread risk of both viruses in black mulberry plantations and the necessity of taking precautions.