Green Synthesis and Characterization of Silver Nanoparticles Using Moringa Peregrina and Their Toxicity on MCF-7 and Caco-2 Human Cancer Cells.

Introduction: The synthesis of nanoparticles using naturally occurring reagents such as vitamins, sugars, plant extracts, biodegradable polymers and microorganisms as reductants and capping agents could be considered attractive for nanotechnology. These syntheses have led to the fabrication of limited number of inorganic nanoparticles. Among the reagents mentioned above, plant-based materials seem to be the best candidates, and they are suitable for large-scale biosynthesis of nanoparticles. Methods: The aqueous extract of Moringa peregrina leaves was used to synthesize silver nanoparticles. The synthesized nanoparticles were characterized by various spectral studies including FT-IR, SEM, HR-TEM and XRD. In addition, the antioxidant activity of the silver nanoparticles was studied viz. DPPH, ABTS, hydroxyl radical scavenging, superoxide radical scavenging, nitric oxide scavenging potential and reducing power with varied concentrations. The anticancer potential of the nanoparticles was also studied against MCF-7 and Caco-2 cancer cell lines. Results: The results showed that silver nanoparticles displayed strong antioxidant activity compared with gallic acid. Furthermore, the anticancer potential of the nanoparticles against MCF-7 and Caco-2 in comparison with the standard Doxorubicin revealed that the silver nanoparticles produced significant toxic effects against the studied cancer cell lines with the IC50 values of 41.59 (Caco-2) and 26.93 (MCF-7) µg/mL. Conclusion: In conclusion, the biosynthesized nanoparticles using M. peregrina leaf aqueous extract as a reducing agent showed good antioxidant and anticancer potential on human cancer cells and can be used in biological applications.

Synergistic cardioprotective ability of co-administration of Moringa supplemented diets and acarbose in diabetic cardiomyopathy involves attenuation of cholinergic, purinergic, monoaminergic, renin-angiotensin system, and antioxidant pathways.

One of the major complications of diabetes mellitus (DM) is diabetic cardiomyopathy (DCM) due to the multifaceted therapy involved. Here, we evaluated the combinatorial effect of Moringa leaf (ML) and seed (MS) supplemented diets plus acarbose (ACA) on cardiac acetylcholinesterase (AChE), adenosine triphosphatase (ATPase), adenosine deaminase (ADA), monoamine oxidase (MAO), arginase, angiotensin-I converting enzyme (ACE), and lactate dehydrogenase (LDH) activities, thiobarbituric acid reactive species (TBARS), and thiols levels. The diets and ACA (25 mg/kg) were administered for 14 days. The fasting blood glucose level (FBGL), cardiac AChE, ATPase, ADA, MAO, arginase, ACE, LDH activities, and TBARS and thiol levels were determined. Relative to the normal rats, the biomarkers were significantly increased in DM rats but were suppressed significantly in the diets plus ACA-treated rats while improving antioxidant status, with the 4% Moringa plus ACA proving outstanding compared to individual ML/MS and ACA. In addition, ML-supplemented diets with/without ACA had better effects compared to MS with/without ACA, respectively. In conclusion, the combination of ML/MS supplemented diets and ACA synergistically modulates the tested biochemicals. However, the effect on blood vessels and the nerves that control the heart, stiffness of left ventricular (LV) hypertrophy, fibrosis, cell signaling abnormalities, related gene expression, clinical trials, and echocardiology studies should be further investigated to affirm this claim. PRACTICAL APPLICATIONS: Moringa oleifera has been a vocal appetite in mitigating cardiovascular disease induced by diabetes, but the formulation of a medicinal diet as an ameliorative route of attention to the pathology is fairly addressed, not talking of its combination with the synthetic antidiabetic drug, such as ACA. Based on this experiment, it is imperative to explore such an idea. This research shows that co-administration of moringa leaf/seed formulated diets plus ACA exhibits a synergistic effect in DCM management. However, further research is needed in this field of experiment.

[Effects of spicy leaves on cognitive function and apoptosis of hippocampal neurons in diabetic rats].

Objective: To investigate the effects of Moringa leaves on the cognitive dysfunction and apoptosis of hippocampal neurons in diabetic rats induced by streptozotocin (STZ). Methods: Fifty male SD rats were selected, and 10 rats were randomly selected as the control group. The other 40 rats were treated with STZ at the dose of 25 mg/kg by intraperitoneal injection. The 40 diabetic rats were randomly divided into model group, Moringa oleifera low-dose, medium-dose and high-dose group. The rats in Moringa oleifera groups were treated with Moringa oleifera at the doses of 2.0, 4.0 and 8.0 g/kg by gavage, the control group and model group were treated with the same amount of normal saline once a day, for 8 weeks. Morris water maze test was used to evaluate the learning and memory ability of rats. Pathological changes of hippocampal neurons and expressions of Bax, caspase-3 and bcl-2 protein in each group were observed by the sections were stained with HE staining and immunohistochemistry. Enzyme linked immunosorbent assay (ELISA) was used to detect tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in rat. Results: compared with the control group, the blood glucose of the model group was increased significantly (P＜0.01), and the blood insulin level was decreased significantly (P＜0.05); compared with the model group, the blood glucose values of Moringa oleifera groups were decreased significantly (P＜0.05, P＜0.01), and the blood insulin levels of middle and high dose Moringa oleifera group were increased significantly (P＜0.05, P＜0.01). There was no significant difference in FBG and INS among the three groups (P＞0.05). In Morris water maze test, compared with the model group, the latency of Moringa oleifera groups was significantly shorter (P＜0.05); the residence time in target quadrant of Moringa oleifera groups with different doses was significantly prolonged (P＜0.05). Compared with the model group, the contents of TNF - α, IL-6 and protein expression in low, medium and high dose groups of Moringa oleifera were decreased significantly (P＜0.05). HE staining and immunohistochemical staining results showed that Moringa oleifera medium dose group was positive, brown yellow, fine granular, compared with the model group. The number of neuronal apoptosis was significantly reduced in the middle dose group (53.21±7.19,P＜0.01); the protein expressions of Bax, caspase-3 and the ratio of Bax/Bcl-2 in hippocampus were significantly decreased in the middle dose group (P＜0.05). Conclusion: The mechanisms of Moringa leaves on the cognitive dysfunction and apoptosis of hippocampal neurons may be related to regulating the protein expressions of Bax, Bcl-2 and Caspase-3, reducing the contents of inflammatory factors TNF-α and IL-6.

Sequenced application of glutathione as an antioxidant with an organic biostimulant improves physiological and metabolic adaptation to salinity in wheat.

Globally, salinity threatens the agricultural crops productivity by inhibiting plant growth and development through osmotic stress and ionic cytotoxicity. The polygenic nature of salinity offers several pragmatic shotgun approaches to improve salinity tolerance. The present study investigated the potential of glutathione (GSH; 1 mM) as an antioxidant and moringa leaf extract (MLE; 3%) as an organic biostimulant applied in sequence as seed priming and foliar spray on wheat growth, physiology and metabolic adaptation under saline conditions (9.16 dS m-1). Plants without any treatment and water spray (H2O) were considered controls. Salinity induced osmotic stress reduced the plant tissue water status and photosynthetic performance, and perturbed ionic (K+/Na+, Ca2+/Na+, K++Ca2+/Na+) and hormonal (IAA, GA3, zeatin, ABA) homeostasis, consequently affected growth and yield in wheat. Sequenced applied MLE and/or GSH improved osmotic stress tolerance by stabilizing membrane integrity and decreasing electrolyte leakage. These positive results were owed to enhanced endogenous GSH and ascorbate levels. Improved tissue water status was attributed to increased osmotic adjustment, better ionic and hormonal homeostasis contributed to improving photosynthetic efficiency and growth under salinity. Exogenously applied MLE and GSH sequences improved grain yield, which was attributed to the maintenance of green leaf area and delayed senescence associated with an increase in photosynthetic pigments and chlorophyll fluorescence traits. In crux, exogenous applied MLE and/or GSH can be the best physiological strategy to reduce the deleterious effects of salinity and improve physiological and metabolic adaptation in wheat under saline field conditions.

In vitro bioaccessibility and bioavailability of iron from fenugreek, baobab and moringa.

Iron deficiency anaemia (IDA) is a common nutritional disorder worldwide. Sustainable food-based approaches are being advocated to use high and bioavailable dietary iron sources to prevent iron deficiency. The study investigated the bioaccessibility and bioavailability of iron from some plant products. Total iron levels in the samples were measured by inductively coupled plasma optical emission spectrometry (ICP-OES). Fractionation of the iron from the digested extracts was carried out by centrifugation and ultrafiltration. Iron bioavailability was determined using an in vitro simulated peptic-pancreatic digestion, followed by measurement of ferritin in Caco-2 cells. The highest amount of bioaccessible iron was obtained from moringa leaves (9.88% ± 0.45 and 8.44 ± 0.01 mg/100 g), but the highest percentage bioavailability was from baobab fruit pulp (99.7% ± 0.13 and 1.74 ± 0.01 mg/100 g) respectively. All the plant products, except for baobab, significantly inhibited iron uptake from FeSO4 and FAC, with fenugreek sprout being the most inhibitory.

Short communication: Effects of moringa extract on adhesion and invasion of Escherichia coli O55 in bovine mammary epithelial cells.

The objective of this study was to evaluate the antibacterial activities of extract derived from moringa leaves. In particular, the effect of moringa extract (Mor) on adhesion and invasion of Escherichia coli O55, Enterococcus faecalis, Staphylococcus simulans, and Serratia liquefaciens was evaluated in bovine mammary epithelial cells (MAC-T). Broth microdilution method, minimum inhibitory concentration and minimum bactericidal concentration assays, adhesion and invasion assays, and real-time PCR were performed. The minimum inhibitory concentration and minimum bactericidal concentration of Mor ranged from 12.5 to 50 mg/mL on 18 out of 27 tested isolates. Treatment of E. coli O55 with Mor (100 and 200 µg/mL) inhibited the adhesion and invasion on MAC-T cells via downregulation of adhesion factors (i.e., papC, f17c-A, and eaeA). Also, when MAC-T cells were pretreated with Mor (200 µg/mL, 12 h) and incubated with E. coli O55, Enterococcus faecalis, Staphylococcus simulans, or Serratia liquefaciens, both E. coli O55 and Enterococcus faecalis showed a significant decrease in adhesion and invasion. Staphylococcus simulans exhibited decreased adhesion and increased invasion. Serratia liquefaciens showed increased adhesion and decreased invasion. In addition, Mor increased mRNA expression of antioxidant enzymes (e.g., heme oxygenase-1, NAD(P)H:quinone oxidoreductase-1, and thioredoxin reductase 1) in MAC-T cells. In conclusion, 12.5 to 50 mg/mL of Mor exhibited antibacterial activity against 18 out of 27 tested isolates. Also, pretreatment of 200 µg/mL of Mor to MAC-T cells modulated adhesion and invasion of E. coli O55 and other mastitis-associated pathogens. Furthermore, Mor increased antioxidant capacities in MAC-T cells, but further in vivo studies are needed.

Moringin and Its Structural Analogues as Slow H2S Donors: Their Mechanisms and Bioactivity.

Moringin (rhamnobenzyl isothiocyanate) is a major bioactive compound in moringa seeds, which have been used as a healthy food. However, its bioactivity mechanisms are not well understood. We investigated moringin and its structurally similar analogues, including benzyl isothiocyanate and 4-hydroxylbenzyl isothiocyanate, for their hydrogen sulfide (H2S)-releasing activity triggered by cysteine. These isothiocyanates rapidly formed cysteine adducts, which underwent intramolecular cyclization followed by slowly releasing (a) organic amine and raphanusamic acid and (b) H2S and 2-carbylamino-4,5-dihydrothiazole-4-carboxylic acids. The product distributions are highly dependent on para-substituents on the phenyl group. Moringin has higher cytotoxicity to cancer cells and is a more potent anti-inflammatory agent than benzyl and hydroxybenzyl analogues, while benzyl isothiocyanate is a better antibacterial agent. Taken together, their bioactivity may not be directly related to their H2S donation activity. However, other metabolites alone do not have cytotoxicity and anti-inflammatory activity. These findings indicated that their activity may be the combination effects of different metabolites via competitive pathways as well the para-substituent groups of benzyl ITCs.

[Functional orientation of Moringa leaves based on text mining and molecular docking technology].

A new method on functional orientation of Moringa leaves based on text mining and molecular docking was explored in the study. First, PubMedplus was used to analyze research data on Moringa leaves collected in PubMed and the indications of Moringa leaves were screened along with the hotspots and development tendency of Moringa leaves. Second, Arrowsmith was used to obtain the biological targets of Moringa leaves. Third, active candidate components of Moringa leaves were filtered by SwissADME analyzing on chemical data collected from literatures. Subsequently, molecular docking between active candidate component and target was studied by systemsDock to forecast the potential active components and their possible effective targets, and GO functional annotation of the potential targets was performed by DAVID database. According to the results, tumor, diabetes and digestive diseases were suggested to be indications of Moringa leaves, correlated with 25 active components and 12 potential effective targets possibly by adjusting G protein-coupled receptor and affecting on inflammatory reaction. The new method on functional orientation by combining text mining with molecular docking was successfully practiced on Moringa leaves as a case study,which provides a useful reference for the ultilization of foreign medicinal resource.

Exploration of arabinogalactan of gum polysaccharide potential in hydrogel formation and controlled drug delivery applications.

Chronic diarrhea is the most common problem in most of the countries with low socio-economic conditions. Hence, efficient therapeutic formulations are required. The present article explores the potential of the gum polysaccharide that itself has an anti-diarrheal activity, to develop the antibiotic drug 'meropenem' carrier, to improve the pharmacotherapy of the diarrhea. The gum based pure and sterile polymeric drug delivery device was prepared by radiation induced crosslinking method. Polymer matrix was characterized by cryo-SEM, AFM, FTIR, 13C NMR, swelling and drug release studies, gel strength, along with some biomedical properties. The slow release was found without a burst effect with non-Fickian diffusion mechanism. It has also been found that polymer was having pore size = 21.73 nm and crosslink density = 5.28 × 10-5mol cm-3 which were synthesized with [HEMA] 4.70 × 10-1 mol/L and irradiation dose 24.62 kGy during copolymerization reaction. The arabinogalactan crosslinked poly(HEMA) polymers were biocompatible, antioxidant and mucoadhesive.

Induction of sub-G0 arrest and apoptosis by seed extract of Moringa peregrina (Forssk.) Fiori in cervical and prostate cancer cell lines.

OBJECTIVE: This study investigated cytotoxicity and induction of apoptosis in human cervical cancer cells (HELA) and prostate cancer cells (PC-3) using the most active fraction of Moringa peregrina seed extract. METHODS: Dried and powdered seeds were extracted using 95% ethanol. The total ethanolic extract was further dissolved in distilled water and separated into petroleum ether, chloroform, ethyl acetate and aqueous extracts. Based on the results of in vitro anticancer studies of all extracts, the most highly active extract was selected for evaluation of apoptosis induction and cell cycle analysis on HELA and PC-3 cells at its half maximal inhibitory concentration using flow cytometry; DNA fragmentation by agarose gel electrophoresis and the expression of protein were measured by Western blot. RESULTS: The chloroform fraction from the ethanolic extract of M. peregrina (CFEE) was the most active antitumor fraction. The selectivity index, determined using the normal Vero cell line, indicated that CFEE had a high degree of selectivity against HELA and PC-3 cells. CFEE induced apoptosis, confirmed by cell cycle arrest at sub-G0 phase and DNA fragmentation. CFEE induced an increase in mRNA expression of caspase-3, a decrease in Bcl-2 mRNA expression, and decreased ATP levels. CFEE increased protein expression of caspase-3 and decreased protein expression of poly-ADP-ribose polymerase-1 (PARP-1). Flow cytometric analysis showed an appreciable increase in the number of cells in the early apoptotic stage in CFEE-treated HELA and PC-3 cells. CFEE treatment significantly increased lipid peroxidation (malondialdehyde level) in HELA and PC-3 cells. CONCLUSION: Seed extract of M. peregrina displayed a significant antitumor effect through apoptosis induction in HELA and PC-3 cells.

Pharmacokinetic of gastrodigenin rhamnopyranoside from Moringa seeds in rodents.

Gastrodigenin rhamnopyranoside (GR) is a hepatoprotective compound that exists in Moringa oleifera seeds. However, the UPLC-MS/MS method for the determination of GR (in-vitro/in-vivo) is lacking clarification. Herein, this study established the UPLC-MS/MS technique, which was effective and sensitive for the investigation of the pharmacokinetics and biodistribution of GR in rats and mice. The separation was achieved with a Shim-pack XR-ODS III C18 column (2.0 × 75 mm, 1.6 µm) at 40 °C, while the mobile phase (Acetonitrile/0.1% Formic acid =12:82, v/v) was at an eluting rate of 0.2 mL/min. The Multiple Reaction Monitoring (MRM) was selected for quantification, i.e., m/z [M + HCOO]- 314.9 → 269 for GR and m/z [M + HCOO] - 182.85 → 137 for Tyrosol as the internal standard. The calibration curves were linearly ranged from 10 to 2500 ng/mL (r ≥ 0.999) with a lower-limit-of-quantification (LLOQ) of 10 ng/mL in the various biological samples (plasma, liver, heart, lung, spleen, brain, kidney). The intra- and inter-day precision was within 5%, while accuracy ranged from -11.4% - 8.33%. Recovery and matrix effect were with 80.32 to 101.31% and 90.36 to 103.76%, respectively, in a reasonable range. After oral and intravenous administration, GR was detected within 3 h but decreased rapidly in plasma, indicating fast elimination. Also, GR was quickly distributed in the various tissues, particularly in the kidney and spleen. The results demonstrated that the established UPLC-MS/MS method was highly linear, precise and accurate with the potential to be used for the quantitative analysis of GR in-vivo.

A preliminary study of fonio-moringa seed meal-based complementary food in Wistar rats.

Effect of fonio-moringa seed meal (FMSM)-based complementary food in Wistar rats was assessed in a 28 days balanced study. Seventy, 21-day-old Wistar rats were allotted to seven groups in a completely randomized design. Infant weaning foods (IWFs) 1, 2, 3, 4, and 5 had 0%, 5%, 10%, 15%, and 20% FMSM inclusion levels, respectively, while two commercial IWFs purchased were coded CFT and CFC. The water absorption capacity, swelling power, and the pasting properties, except peak time varied (p < .05) among the IWFs. Rats fed on IWFs 3 and 4 had comparable weight gain with those fed on the commercial foods. The heart and kidney relative weights were influenced (p < .05), while villus length (duodenum) of the rats were significant (p < .05). The blood indices were not significant, but the alanine aminotransferase and cholesterol levels ranged 15.65-32.25 µ/dl and 75.75-94.55 mg/100 ml, respectively. Incorporation of 10% FMSM is recommended in IWFs. PRACTICAL APPLICATIONS: The need to reduce hunger and starvation among the less-privileged people in society is becoming increasingly important. Specifically, many infants, preschool and school children are often not having access to nutritive foods that will enhance their mental alertness. The commercial IWFs on sale in Nigeria are in most cases high in prices and consequently out of the reach of the populace. Fonio is a starchy grain with an important potential not only as a survival food but as a compliment for standard diets. Also, moringa seed is known to be of nutritional value. Most infant formulae are based on maize and soybean and it is hoped that the incorporation of FMSM in the infant weaning formula will help to increase the nutritive value and stem the cost of IWFs.

Partial ameliorative effect of Moringa leaf ethanolic extract on the reproductive toxicity and the expression of steroidogenic genes induced by subchronic cadmium in male rats.

The impact of Moringa oleifera leaf ethanol extract (MOLEE) was assessed on the expression of the steroidogenic genes (steroidogenic acute regulatory protein (StAR) and cytochrome P450c17 subfamily a (CYP17a) and luteinizing hormone receptor (LHR) gene) as well as on the cadmium chloride (CdCl2)-induced reproductive toxicity for 56 days in male rats. Four groups were used: control, Moringa-treated (MOLEE), CdCl2-treated, and CdCl2 + MOLEE groups. The reproductive toxicity of CdCl2 was confirmed; it caused a significant decrease in the accessory sex organ weights, testosterone level, testicular GST level, elevated MDA level (lipid peroxidation indicator), and histopathological alterations in seminiferous tubules, prostate, seminal vesicles, and epididymis as well as sperm characteristics. It also induced downregulation in the expression of StAR and CYP17a genes without change in the expression LHR gene. Eleven active compounds were detected in the GC-MS analysis of MOLEE; six of them have antioxidant properties, and five new compounds presented variable activities. MOLEE alone induced a stimulatory effect on the expression of steroidogenic and LHR genes. It restored the weight of reproductive organs to the control level; however, the recovery in sperm count, motility, abnormalities, percentage of alive sperm, testosterone, and MDA level are still comparable with the control level. Similar findings were also reported at the histological structure of the testes, epididymis, and accessory sex glands. Complete recovery of the GST enzyme activity was observed. Additionally, a restoration in the expression level of the steroidogenic genes was also reported. Our results indicated that the concurrent administration of MOLEE with CdCl2 can partially mitigate its harmful effects on male fertility.

CpG methyl-seq and RNA-seq epigenomic and transcriptomic studies on the preventive effects of Moringa isothiocyanate in mouse epidermal JB6 cells induced by the tumor promoter TPA.

Epigenetic mechanisms play an important role in the early stages of carcinogenesis. Moringa isothiocyanate (MIC-1) is a major bioactive component derived from Moringa oleifera that has considerable antioxidant and anti-inflammatory effects. However, how MIC-1 influences epigenomic alterations in TPA-mediated JB6 cell carcinogenic transformation has not been evaluated. In this study, DNA and RNA isolated from TPA-induced JB6 cells in the presence or absence of MIC-1 were subjected to DNA Methyl-seq and RNA-seq to identify differentially methylated regions (DMRs) and differentially expressed genes (DEGs), respectively. When JB6 cells were challenged with TPA alone, there was a significant alteration of DEGs and DMRs; importantly, MIC-1 treatment reversed the patterns of some of the DEGs and DMRs. Transcriptome and CpG methylome profiling was performed in Ingenuity® Pathway Analysis (IPA) software to analyze the altered signaling pathways. Several anti-inflammatory responses, antioxidative stress-related pathways, and anticancer-related pathways were identified to be affected by MIC-1. These pathways included NF-kB, IL-1, LPS/IL-1-mediated inhibition of RXR function, Nrf2-mediated oxidative stress response, p53, and PTEN signaling pathways. Examination of correlations between transcriptomic and CpG methylome profiles yielded a small subset of genes, including the cancer-related genes Tmpt, Tubb3, and Muc2; the GTPases Gchfr and Igtp; and the cell cycle-related gene Cdc7. Taken together, our results show the potential contributions of epigenomic changes in DNA CpG methylation to gene expression to molecular pathways active in TPA-induced JB6 cells and demonstrate that MIC-1 can reverse these changes, supporting the potential preventive/treatment effects of MIC-1 against skin carcinogenesis.

Moringa peregrina Leaves Extracts Induce Apoptosis and Cell Cycle Arrest of Hepatocellular Carcinoma.

Moringa grows in the tropical and subtropical regions of the world. The genus Moringa belongs to family Moringaceae. It is found to possess various medicinal uses including hypoglycemic, analgesic, anti-inflammatory, hypolipidemic, and antioxidant activities. In this study, we investigated the antimicrobial and the anticancer activity of the Moringa peregrina as well as Moringa oleifera leaves extracts grown locally in Egypt. Results indicated that most of the extracts were found to possess high antimicrobial activity against gram-positive bacteria, gram-negative bacteria, and fungus. The survival rate of cancer cells was decreased in both hepatocellular carcinoma (HepG2) and breast carcinoma (MCF-7) cell lines when treated with Moringa leaves extracts. In addition, the cell cycle progression, apoptosis, and cancer-related genes confirmed its anticancer effect. The toxicity of each extract was also tested using the normal melanocytes cell line HFB4. The toxicity was low in both Moringa peregrina and Moringa oleifera leaves extracts. Furthermore, GC/MS analysis fractionized the phytochemicals content for each potential extract. In conclusion, results suggested that the Moringa peregrina and Moringa oleifera leaves extracts possess antimicrobial and anticancer properties which could be attributed to the bioactive phytochemical compounds present inside the extracts from this plant. These findings can be used to develop new drugs, especially for liver cancer chemotherapy.

Moringa Isothiocyanate Activates Nrf2: Potential Role in Diabetic Nephropathy.

Moringa isothiocyanate (MIC-1) is the main active isothiocyanate found in Moringa oleifera, a plant consumed as diet and traditional herbal medicine. Compared to sulforaphane (SFN), MICs are less studied and most work have focused on its anti-inflammatory activity. The purpose of this study is to better understand the Nrf2-ARE antioxidant activity of MIC-1 and its potential in diabetic nephropathy. MIC-1 showed little toxicity from 1.25-5 µM. MIC-1 activated Nrf2-ARE at similar levels to SFN. MIC-1 also increased gene expression of downstream Nrf2 genes NQO1, HO-1, and GCLC. Protein expression of HO-1 and GCLC was elevated in MIC-1-treated cells versus control. MIC-1 suppressed pro-inflammatory cytokines in LPS-stimulated macrophages. MIC-1 reduced levels of reactive oxygen species in high glucose (HG)-treated human renal proximal tubule HK-2 cells. RNA-seq was performed to examine the transcriptome in HK-2 cells exposed to HG with or without MIC-1. Ingenuity Pathway Analysis (IPA) of RNA-seq on HK-2 cells exposed to HG identified TGFß1 and NQO1 regulation as potentially impacted and treatment of HG-exposed HK-2 cells with MIC-1 reversed the gene expression of these two pathways. Results implicate that the transcriptional regulator TGFß1 signaling is activated by HG and that MIC-1 can inhibit HG-stimulated TGFß1 activation. In summary, MIC-1 activates Nrf2-ARE signaling, increases expression of Nrf2 target genes, and suppresses inflammation, while also reducing oxidative stress and possibly TGFß1 signaling in high glucose induced renal cells. Taken together, it appears that one potential therapeutic strategy for managing DN and is currently under development in clinic is Nrf2 activation.

Radiation-induced graft copolymerization of N­vinyl imidazole onto moringa gum polysaccharide for making hydrogels for biomedical applications.

Keeping in view the importance of polysaccharide gum in the pharmaceutical formulations, in the present work, exploration of the potential of the moringa gum in hydrogel formation for drug delivery applications has been carried out. The gum based hydrogels were prepared via radiation induced graft-copolymerization of N­vinyl imidazole onto the gum. The polymers were characterized by cryo-SEM, AFM, FTIR, 13C NMR spectroscopy and swelling studies. Some properties of the polymers such as blood compatibility, antioxidant activity, and mucoadhesion and gel strength were also determined along with the evaluation of the drug release profile of an antibiotic drug levofloxacin. The slow release of drug was observed without burst effect from the drug loaded hydrogels. Release of the drug followed non-Fickian diffusion mechanism and release profile was best fitted in in Hixson-Crowell kinetic model. Cryo-SEM showed the porous nature of the hydrogels and AFM analysis confirmed the surface roughness. The polymers were found to be non-haemolytic mucoadhesive and antioxidant in nature with pore size = 12.09 nm and crosslinking density = 9.13 × 10-5 mol cm-3. These results indicated that these pure and sterile polymers can be proposed as a gastrointestinal drug delivery system.

Hydrogel formation by radiation induced crosslinked copolymerization of acrylamide onto moringa gum for use in drug delivery applications.

Keeping in view the importance of polysaccharides gum in designing drug delivery systems, the present work is the exploration of the potential of the moringa gum in hydrogel formation via radiation induced crosslinking method for drug delivery applications. These polymers were characterized by cryo-SEM, AFM, FTIR, 13C-NMR spectroscopy and swelling studies. Some properties of the polymers such as blood compatibility, antioxidant activity, mucoadhesion and gel strength were also determined along with the evaluation of drug release profile of an antibiotic drug levofloxacin. The slow release of drug was observed without burst effect from the drug loaded hydrogels. Release of drug occurred through non-Fickian diffusion mechanism and release profile best fitted in Korsmeyer-Peppas kinetic model. Cryo-SEM showed the porous nature of the hydrogels. The polymers were found to be mucoadhesive and antioxidant in nature. These results indicated that these pure and sterile polymers can be proposed as gastrointestinal drug delivery system.

The Diversity of Chemoprotective Glucosinolates in Moringaceae (Moringa spp.).

Glucosinolates (GS) are metabolized to isothiocyanates that may enhance human healthspan by protecting against a variety of chronic diseases. Moringa oleifera, the drumstick tree, produces unique GS but little is known about GS variation within M. oleifera, and even less in the 12 other Moringa species, some of which are very rare. We assess leaf, seed, stem, and leaf gland exudate GS content of 12 of the 13 known Moringa species. We describe 2 previously unidentified GS as major components of 6 species, reporting on the presence of simple alkyl GS in 4 species, which are dominant in M. longituba. We document potent chemoprotective potential in 11 of 12 species, and measure the cytoprotective activity of 6 purified GS in several cell lines. Some of the unique GS rank with the most powerful known inducers of the phase 2 cytoprotective response. Although extracts of most species induced a robust phase 2 cytoprotective response in cultured cells, one was very low (M. longituba), and by far the highest was M. arborea, a very rare and poorly known species. Our results underscore the importance of Moringa as a chemoprotective resource and the need to survey and conserve its interspecific diversity.

Anti-inflammatory and analgesic activities of solvent fractions of the leaves of Moringa stenopetala Bak. (Moringaceae) in mice models.

BACKGROUND: Many people still experience pain and inflammation regardless of the available drugs for treatments. In addition, the available drugs have many side effects, which necessitated a quest for new drugs from several sources in which medicinal plants are the major one. This study evaluated the analgesic and anti- inflammatory activity of the solvent fractions of Moringa stenopetala in rodent models of pain and inflammation. METHODS: Successive soxhlet and maceration were used as methods of extractions using solvents of increasing polarity; chloroform, methanol and water. Swiss albino mice models were used in radiant tail flick latency, acetic acid induced writhing and carrageenan induced paw edema to assess the analgesic and anti-inflammatory activities. The test groups received different doses (100 mg/kg, 200 mg/kg and 400 mg/kg) of the three fractions (chloroform, methanol and aqueous). The positive control groups received morphine (20 mg/kg) or aspirin (100 mg/kg or 150 mg/kg) based on the respective models. The negative control groups received the 10 ml/kg of vehicles (distilled water or 2% Tween 80). RESULTS: In all models, the chloroform fraction had protections only at a dose of 400 mg/kg. However, the methanol and aqueous fraction at all doses have shown significant central and peripheral analgesic activities with a comparable result to the standards. The aqueous and methanol fractions significantly reduced carrageenan induced inflammation in a dose dependent manner, in which the highest reduction of inflammation was observed in aqueous fraction at 400 mg/kg. CONCLUSION: This study provided evidence on the traditionally claimed uses of the plant in pain and inflammatory diseases, and Moringa stenopetala could be potential source for development of new analgesic and anti-inflammatory drugs.