Larrea divaricata a native Argentine plant with potential activity against SARS-CoV-2? / ¿Larrea divaricata una planta nativa argentina con actividad potencial sobre SARS-CoV-2?

Larrea divaricata Cav. is an autochthonous South American plant popularly used in inflammatory and infectious diseases with reported anti - inflammatory, immunomodulatory, antimicrobial and antioxidant activities. Covid - 19 is an infection ca used by the severe acute respiratory syndrome coronavirus 2 (SARS - CoV - 2). This virus can cause pneumonia and even death in about 5% of the cases. The objective of the article was to demonstrate, through a literature review, that L. divaricata has sufficie nt attributes to be assayed against SARS - CoV - 2. For this, the chemical composition, reported activities and docking studies were taken into account. This review demonstrated that the plant extracts are capable of inhibiting the proliferation of fungi, bact eria and viruses and that they exert anti - inflammatory and immunomodulatory actions in different " in vitro " and " in vivo " models. These results suggest that the plant is a good candidate to be studied for the prevention and/or treatment of SARS - CoV - 2.

Larrea divaricata Cav. es una planta autóctona Sudamericana, utilizada popularmente en enfermedades inflamatorias e infecciosas, con activida d anti - inflamatoria, inmunomoduladora, antimicrobiana y antioxidante reportada. El Covid - 19 es una infección causada por una cepa de coronavirus, SARS - CoV - 2 (coronavirus tipo 2 causante del síndrome respiratorio agudo severo). Este virus puede originar neu monía e incluso la muerte en alrededor del 5% de los casos. Nuestro objetivo fue demostrar, a través de una revisión bibliográfica, que esta planta tiene atributos suficientes para ser ensayada en estudios contra SARS - CoV - 2. Se tuvo en cuenta la composici ón química, los antecedentes científicos y los estudios de acoplamiento molecular. Esta revisión permitió demostrar que extractos de la planta son capaces de inhibir la proliferación de hongos, bacterias y virus y que presentan acción anti - inflamatoria en diferentes modelos " in vitro " e " in vivo ", lo que los hace candidatos a ser estudiados en la prevención y/o tratamiento de la infección contra SARS - CoV - 2.

New insights into the antiviral activity of nordihydroguaiaretic acid: Inhibition of dengue virus serotype 1 replication.

BACKGROUND: Dengue virus (DENV) is considered one of the most important pathogens in the world causing 390 million infections each year. Currently, the development of vaccines against DENV presents some shortcomings and there is no antiviral therapy available for its infection. An important challenge is that both treatments and vaccines must be effective against all four DENV serotypes. Nordihydroguaiaretic acid (NDGA), isolated from Larrea divaricata Cav. (Zygophyllaceae) has shown a significant inhibitory effect on a broad spectrum of viruses, including DENV serotypes 2 and 4. PURPOSE: We evaluated the in vitro virucidal and antiviral activity of NDGA on DENV serotype 1 (DENV1), including the study of its mechanism of action, to provide more evidence on its antiviral activity. METHODS: The viability of viral particles was quantified by the plaque-forming unit reduction method. NDGA effects on DENV1 genome and viral proteins were evaluated by qPCR and immunofluorescence, respectively. Lysosomotropic activity was assayed using acridine orange and neutral red dyes. RESULTS: NDGA showed in vitro virucidal and antiviral activity against DENV1. The antiviral effect would be effective within the first 2 h after viral internalization, when the uncoating process takes place. In addition, we determined by qPCR that NDGA decreases the amount of intracellular RNA of DENV1 and, by immunofluorescence, the number of cells infected. These results indicate that the antiviral effect of NDGA would have an intracellular mechanism of action, which is consistent with its ability to be incorporated into host cells. Considering the inhibitory activity of NDGA on the cellular lipid metabolism, we compared the antiviral effect of two inhibitors acting on two different pathways of this type of metabolism: 1) resveratrol that inhibits the sterol regulatory element of binding proteins, and 2) caffeic acid that inhibits the 5-lipoxygenase (5-LOX) enzyme. Only caffeic acid produced an inhibitory effect on DENV1 infection. We studied the lysosomotropic activity of NDGA on host cells and found, for the first time, that this compound inhibited the acidification of cell vesicles which would prevent DENV1 uncoating process. CONCLUSION: The present work contributes to the knowledge of NDGA activity on DENV. We describe its activity on DENV1, a serotype different to those that have been already reported. Moreover, we provide evidence on which stage/s of the viral replication cycle NDGA exerts its effects. We suggest that the mechanism of action of NDGA on DENV1 is related to its lysosomotropic effect, which inhibits the viral uncoating process.

Anti-hyperglycaemic effect and nutritional properties of an aqueous extract of Larrea divaricata Cav. (jarilla) in streptozotocin-induced diabetes in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Larrea divaricata Cav. (Zygophyllaceae) (jarilla) is a native plant of South America widely distributed across Argentina and used in popular medicine to treat diabetes and hypercholesterolemia by the Diaguita-Calchaquí, Amaichas, and Quilmes indigenous communities and by non-indigenous population (criollos) of Calamuchita, in the province of Córdoba, Argentina. L. divaricata has also proved to have anti-inflammatory properties. However, the antidiabetic effects and the nutritional properties of the aqueous extract (AE) of this plant remain to be scientifically determined. AIM OF THE STUDY: The aim of the present work was to evaluate the capacity of an aqueous extract of L. divaricata (AE) and its main compound nordihydroguaiaretic acid (NDGA) to modulate the glucose, cholesterol, triglycerides and oxidative stress levels in STZ-induced diabetes in mice. The general objective of the present work was to search for extracts that can be used as adjuvant therapy in for diabetes. The suitability of the extract to be used as a dietary supplement was also assessed by determining the proximate amount of fibre, lipids, proteins, and minerals. MATERIALS AND METHODS: Diabetes was induced in mice by administration of streptozotocin (STZ). AE and NDGA were administered by the oral route. The animals' glycaemia was periodically monitored in blood samples obtained from the tail vein. The glucose dehydrogenase method was used. The effect of the AE on cholesterol, triglycerides, oxidative stress, lipid peroxidation and reduced glutathione (GSH) levels were determined in plasma samples by spectrophotometric assays. RESULTS: In STZ-treated mice, AE significantly decreased glucose (33%, ****p < 0.0001) and cholesterol levels (32%, **p < 0.01). AE and NDGA decreased lipid peroxidation (30% and 38%, respectively, ****p < 0.0001), and increased GSH levels (20%, **p < 0.01). The effects of AE on glucose and lipid levels could not be ascribed to NDGA; however, this compound was involved in the extract antioxidant effects. The overall effects of AE were probably related to its antioxidant activity and to the anti-hyperglycaemic effect mainly mediated by flavonoids, fibre (carbohydrates) and mineral elements such as potassium, calcium, magnesium, and zinc. The AE protein content also confers the extract nutritional properties. CONCLUSIONS: These results support the hypothesis that AE could be used as a therapeutic adjuvant or as a nutritional supplement to control glucose levels and lipid metabolism in metabolic syndrome-associated diseases. Moreover, these results scientifically reinforce the popular use of the plant.

Larrea divaricata: anti-inflammatory and antioxidant effects of on macrophages and low density lipoproteins.

BACKGROUND: The oxidized low density lipoprotein (ox-LDL) contributes to inflammation and oxidative stress through the activation of macrophages under hyperglycemia contributing to the development of diabetes mellitus and to atherosclerosis. Plants are a source of effective and innocuous antioxidants. Larrea divaricata Cav. (Zygophyllaceae) is used in Argentina folk medicine for its anti-inflammatory properties. METHODS: The aim of this work was to study the antioxidant and anti-inflammatory effects of the aqueous extract (AE) of L. divaricata on macrophages under glucose stimulation and on human LDL and HDL particles under free radical generators. RESULTS: AE reduced the lipid peroxidation (17%), nitric oxide (NO) (47-50%), tumor necrosis factor-α (TNF-α) (32%) and free radicals (50%) induced by glucose on macrophages. Also prevented HDL nitration (28%), thus preserving its function and structure and inhibited LDL oxidation. The effect on the nitrosative stress was mainly driven by nordihydroguaiaretic acid (NDGA). CONCLUSIONS: These results suggest a potential usefulness of AE as an adjuvant phytotherapy in patients with diabetes mellitus and atherosclerosis.

Antiproliferative activity of aqueous and polyphenol-rich extracts of Larrea divaricata Cav. on a melanoma cell line.

Most of the deaths from skin cancer are caused by melanoma, a malignancy in which STAT3 plays a crucial role. The inhibition of STAT3 is considered a potential target to induce cell death, tumor regression and metastasis inhibition. The objective of this work was to evaluate the activity of the aqueous extract of Larrea divaricata (Aq), a fraction rich in polyphenols (EA),and the isolated compound quercetin-3-methyl ether (Q3ME) on B16F10 melanoma cells. The effects of Aq, EA and Q3ME were assessed on B16F10 cells by determining the proliferation, viability, apoptosis induction and the expression and phosphorylation of STAT3. The phytochemical composition of the extracts was determined by High Performance Liquid Chromatography. Aq, EA and Q3ME presented antiproliferative activity on B6F10 cells through p-STAT3 inhibition and early and late apoptosis induction (EC50 EA= ≤0.1 µg/ml; Aq= 316 ± 30 µg/ml; Q3ME= <0.1 µg/ml). L. divaricata could be considered for the development of adjuvant phytotherapies in melanoma treatment.

UHPLC-ESI-OT-MS Phenolics Profiling, Free Radical Scavenging, Antibacterial and Nematicidal Activities of "Yellow-Brown Resins" from Larrea spp.

This research was designed to investigate the metabolite profiling, phenolics and flavonoids content and the potential antioxidant, antibacterial and nematicidal activities of "yellow-brown resins" from Larrea divaricata Cav (LdRe) and L. nitida Cav (LnRe). Metabolite profiling was obtained using an ultrahigh resolution liquid chromatography orbitrap MS analysis (UHPLC-ESI-OT-MS). The antioxidant properties were screened by four methods: 2,2-diphenyl-1-picrylhydrazyl assay (DPPH), trolox equivalent antioxidant activity assay (TEAC), ferric-reducing antioxidant power assay (FRAP) and lipid peroxidation in erythrocytes (LP). The antibacterial activity was evaluated according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. In addition, the potential combinatory effect was analyzed with the fractional inhibitory concentration index (FICI) values using the checkerboard design. The nematicidal activity was carried out according to a standardized protocol. LdRe and LnRe showed a strong capture of the DPPH radical withvalues around 8.4 µg resin/mL; FRAP (1.69-1.94 mgTE/ g resin), TEAC (1.08-1.09 mgTE/g resin) and LP (81-82% at 100 µg of resin/mL) assays. A strong antimicrobial activity was displayed by both resins against methicillin-sensitive Staphylococcus aureus ATCC 25923(MSSA) and methicillin-resistant S. aureus ATCC 43300(MRSA) (MICs = 16-32 µg resin/mL). Additionally, the combination of LdRe or LnRe with the antibiotic cefotaxime showed an indifferent effect (FICI values = 1-1.25), however, this combinationcould be a potential strategy to reduce the drug doses, and in this way can be a potential alternative to reduce bacterial resistance. On the other hand, the resins showed a scarce nematicidal potential toward J2 Meloidogyne incognita; an important nematode infecting horticultural crops. Phenolics compounds were identified by UHPLC-PDA-OT-MS analysis, updating the knowledge on the chemical profile of these species. These results, together with the high content of quantified phenolics and flavonoids, allow the phenolics-enriched resins of these two Larrea species to be considered as a promising sustainable source of compounds of pharmacological interest.

Antigenotoxic, antiproliferative and antimetastatic properties of a combination of native medicinal plants from Argentina.

ETHNOPHARMACOLOGICAL RELEVANCE: Jarilla is the common name of an appreciated group of native plants from the semi-arid region in Argentina (Larrea cuneifolia Cav., Larrea divaricata Cav. and Zuccagnia punctata Cav.) that have been historically consumed to heal respiratory, musculoskeletal and skin ailments, as well as recommended for weakness/tiredness, hypertension, diabetes and cancer treatment. It was previously reported that some biological properties could be improved when these plants are used jointly. Infusions of a defined mixture, composed by three Jarilla species, L. cuneifolia: L. divaricata: Z. punctata (0.5:0.25:0.25) (HM2) showed synergistic and additive effect on antioxidant activity even after passing through the gastro-duodenal tract. AIM OF THE STUDY: The main purpose of this work was to evaluate antigenotoxic, antitumor, and anti-metastatic properties of the Jarilla species that grow in the Northwest of Argentina and a herbal combination of them. MATERIAL AND METHODS: Infusions of Jarilla mixture (HM2), and of each single plant species were prepared. Phenolic profiles of infusions were analyzed by HPLC-ESI-MS/MS and two relevant chemical markers were quantified. The antigenotoxic activity was evaluated by using the Ames test and the Cytokinesis-Block Micronucleus (CBMN) assay against direct mutagens. Evaluations of both cytotoxicity and antiproliferative effects were conducted on tumor and non-tumor cell lines. Both in vivo tumoral growth and metastasis inhibition were evaluated by using a carcinoma model on Balb/c mice. RESULTS: HM2 mix could suppress genetic and chromosome mutations induced by 4-nitro-o-phenylendiamine (4-NPD) and doxorubicin. Herbal mixture and single plant infusions showed cytotoxic effect against mammary, uterus, and brain tumoral cells without a selective action vs normal human cell line. HM2 mix was able to reduce mammary tumor mass on the Balb/c mice model and showed a significant reduction in the number of metastatic nodules in the lungs. CONCLUSIONS: Our results suggest that the combinations of three Jarilla species from northwest Argentina would be a promising alternative to treat or slow down the development of chronic diseases, such as cancer.

Comprehensive and quantitative profiling of lipid molecular species by LC-ESI-MS/MS of four native species from semiarid Patagonian Monte.

The maintenance of lipid and fatty acids unsaturated composition has been described as one of the mechanisms associated to drought tolerance, but research about the lipid profile in native plants of semiarid environment is still limited. The primary objective was to study whether lipid profiles correlates with drought resistance strategies (tolerant or avoidant) of two life forms (shrubs and grasses). The lipid classes and molecular species of green leaves of Larrea divaricata and Lycium chilense shrubs and Pappostipa speciosa and Poa ligularis grasses were determined using LC-ESI-MS/MS. The soil water content was very low during spring and leaf relative water content was between 47 and 74% in the four species. Lipid profiling was different between both life forms. The prevalent compounds were digalactosyldiacylglycerol (DGDG), monogalactosyldiacylglycerol (MGDG) and phosphatidic acid (PA). The lipid signature shows that L. divaricata adjust its lipid composition to tolerate drought, increasing the content of: a) total lipids and total phospholipids, b) structural phospholipids (36:4 and 36:2-PC, phosphatidylcholine; 36:4-PE, phosphatidylethanolamine), c) chloroplast and mitochondria lipids (32:1 and 32:0-PG, phosphatidylglycerol; 34:3, 36:6 and 36:3-DGDG), d) signaling lipids (34:3, 34:2 and 36:5-PA and PI, phosphatidylinositol), and e) polyunsaturated fatty acids (PUFAs, 18:3 and 18:2) and long chain polyunsaturated fatty acids (LC-PUFAs, in 40:2 and 42:2-PS, phosphatidylserine). This membrane lipid composition contributes to membrane stabilization as metabolic-functional strategy for drought tolerance in the Patagonian Monte. In addition, the 18:3 present in lipids of both grasses could be incorporated to lamb fed based on pastures and result healthy for human dietary.

Molecular mimicry between Larrea divaricata Cav. plant and a reference strain of Pseudomonas aeruginosa.

Currently, the world health sector faces a big problem due to the increase of bacterial strains resistant to antibiotics. In 2017, the World Health Organization reported a list of resistant bacteria, among which Pseudomonas aeruginosa was present. This opportunistic pathogen is associated to nosocomial infections, and no effective vaccines against this bacterium have been found. Larrea divaricata Cav. (jarilla) is a shrub highly distributed in America and widely used in folk medicine. In our laboratory, cross-reactivity of antibodies obtained from the recognition of jarilla proteins against proteins from gram-negative bacteria has been demonstrated. The objective of this study was to study the cross-reactivity of anti-L. divaricata antibodies with P. aeruginosa extracellular proteins in order to find an innocuous prophylactic therapy against this nosocomial pathogen. We observed that antibodies generated by proteins from jarilla crude extract recognized antigenic determinants present in extracellular proteins of P. aeruginosa. However, further studies are needed to investigate the neutralizing capacity of these antibodies on the specific enzymatic proteins involved in the pathogenicity of this bacterium.

Larrea divaricata Cav. aqueous extract and nordihydroguariaretic acid modulate oxidative stress in submandibular glands of diabetic rats: a buccal protective in diabetes.

BACKGROUND: Oxidative stress is an imbalance between the levels of reactive oxygen species (ROS), reactive nitrogen species (RNS) and endogenous antioxidants. The aetiology and pathogenesis of several oral diseases are attributed to this process. The antioxidant enzymes secreted in the saliva by submandibular glands maintain oral health through the scavenging of ROS. The objective of this work was to study the capacity of an aqueous extract of L. divaricata (AE), and its majority compound, nordihydroguariaretic acid (NDGA), to modulate the pro-oxidant/antioxidant status in submandibular glands in a model of oxidative stress induced by streptozotocin (STZ) in rats. METHODS: To induce oxidative stress with STZ, a group of animals was treated i.p. with 1 X PBS (control group) and other group was injected i.p. once with STZ (60 mg/kg). Ten days after the treatment, blood samples were taken from the tail vain to determine the glucose levels. Animals with glucose values ≥300 mg/ml were selected. The submandibular glands of control and STZ treated animals were incubated with either the AE (500 µg/ml) or with NDGA (1.5 µg/ml), and the content of malondialdehyde (MDA), protein carbonyl groups, ROS and RNS, and the activity and expression of peroxidase (Px), superoxide dismutase (SOD) and inducible nitric oxide synthase (iNOS) were assayed. RESULTS: AE decreased the levels of MDA (##P < 0.01) and protein carbonyl groups (#P < 0.05), and modulated the levels of ROS such as hydrogen peroxide (H2O2)(##P < 0.01), superoxide anion (O2.-) (#P < 0.05) and nitric oxide (NO) (#P < 0.05) in relation to the modulation of Px and iNOS expression. NDGA was found to be involved in these effects. CONCLUSIONS: The antioxidant activity of the AE in the submandibular glands would allow the maintenance of the antioxidant pool to prevent oral oxidative diseases.

Enhanced oxidative stability of meat by including tannin-rich leaves of woody plants in goat diet.

Objective: The aim of this study was to evaluate the effect of dietary incorporation of tannin-rich woody species on meat oxidative stability, carcass traits and meat quality in goats. Methods: Two tannin-rich species were tested using a three-treatments feeding trial, where treatments consisted of: Larrea divaricata and Acacia aroma both at 12.5% in dry matter basis of the diet and a control diet (alfalfa hay). All feeding diets were iso-protein and iso-energy. Carcass conformation, carcass compactness, carcass fatness and subcutaneous fat deposition were evaluated. Intake, liveweigh, Longissimus thoracis et lumborum muscles of goats were analyzed in order to evaluate quality parameters such as pH value, instrumental color evaluation, water holding capacity, total phenolic content, antioxidant activity, meat oxidative stability and fatty acid profiles in meat. Results: Feed intake, liveweight gain, carcass and meat traits did not differ among treatments. Changes in meat lipid profile among treatments were observed for oleic and elaidic acid contents. Meat total phenolic content and antioxidant activity did not differ among treatments; although, meat oxidative status after storage at room temperature, as well as under refrigerated and frozen conditions were different between control and both supplemented groups. Conclusion: The inclusion of Acacia aroma and Larrea divaricata leaves in goat diet enhanced meat oxidative stability. Modulation of the ruminal biohydrogenation of fatty acids produced by condensed tannins of these plant species need to be further investigated.

Food preservation by Larrea divaricata extract: participation of polyphenols.

The aim of this study was to evaluate the antioxidant and protease inhibitor capacities on eggs and milk protein of a nor-dihydroguaiaretic (NDGA)-standardized aqueous extract of Larrea divaricata (AE) and to analyze the participation of polyphenols as NDGA in these actions. NDGA was determined by high-performance liquid chromatography; flavonoids and polyphenols were quantified by spectrophotometric methods as well as inhibition of lipid peroxidation, proteinase inhibitor capacity, advanced glycation end products (AGES) formation, and inhibition of albumin denaturation. The extract protected food for oxidative damage by preventing malondialdehyde formation in egg yolk and by preventing AGE formation in completely cooked eggs, also impeded albumin denaturation, and casein hydrolysis induced by trypsin and heat. Polyphenols, especially flavonoids and NDGA, were involved in these actions.

Development of pH-responsive biopolymer-silica composites loaded with Larrea divaricata Cav. extract with antioxidant activity.

A detailed study of biomaterials is mandatory to comprehend their feasible biomedical applications in terms of drug delivery and tissue regeneration. Particularly, mucoadhesive biopolymers such as chitosan (chi) and carboxymethylcellulose (CMC) have become interesting biomaterials regards to their biocompatibility and non-toxicity for oral mucosal drug delivery. In this work, pH-responsive biopolymer-silica composites (Chi-SiO2, Chi-CMC-SiO2) were developed. These two types of composites presented a different swelling behavior due to the environmental pH. Moreover, the nanocomposites were loaded with aqueous Larrea divaricata Cav. extract (Ld), a South American plant which presents antioxidant properties suitable for the treatment of gingivoperiodontal diseases. Chi-CMC-SiO2 composites showed the highest incorporation and reached the 100% of extract release in almost 4 days while they preserved their antioxidant properties. In this study, thermal and swelling behavior were pointed out to show the distinct water-composite interaction and therefore to evaluate their mucoadhesivity. Furthermore, a cytotoxicity test with 3T3 fibroblasts was assessed, showing that in both composites the addition of Larrea divaricata Cav. extract increased fibroblast proliferation. Lastly, preliminary in vitro studies were performed with simulated body fluids. Indeed, SEM-EDS analysis indicated that only chi-SiO2 composite may provide an environment for possible biomineralization while the addition of CMC to the composites discouraged calcium accumulation. In conclusion, the development of bioactive composites could promote the regeneration of periodontal tissue damaged throughout periodontal disease and the presence of silica nanoparticles could provide an environment for biomineralization.

Immunization with Larrea divaricata Cav. proteins elicits opsonic antibodies against Pseudomonas aeruginosa and induces phagocytic activity of murine macrophages.

Pseudomonas aeruginosa is an opportunistic pathogen implicated in nosocomial infections for which no vaccines have been approved. Larrea divaricata Cav. (Jarilla) is a widely spread plant in America and it is used in folk medicine to treat several pathologies. It has also been shown that antibodies elicited against Jarilla proteins of crude extract (JPCE) cross-react with proteins from gram-negative bacteria. In this study we aim to assess the contribution of anti-JPCE antibodies in the opsonophagocytosis of P. aeruginosa by murine macrophages. Levels of reactivity of anti-JPCE IgG and IgA antibodies against cell and membrane proteins suggest that these proteins induce a response that could favor opsonic bacterial recognition, which is important for the elimination of bacteria on mucous membranes, useful in the early stages of infection. Opsonophagocytosis assays also show that these antibodies could favor bacteria intake. These results together with previous observations that indicate that anti-JPCE antibodies are able to neutralize P. aeruginosa enzymes point L. divaricata proteins as candidates for vaccine development.

Stability of an Aqueous Extract of Larrea divaricata Cav. during a Simulated Digestion Process.

Larrea divaricata Cav. (Zygophyllaceae) is a South American plant widely distributed in Argentina that is used in folk medicine to treat inflammatory diseases. The aqueous extract is known to have well-documented biological activities such as antitumour, immunomodulatory, antimicrobial, antiinflammatory and antioxidant. However, its stability in gastrointestinal fluids is unknown. The latter is an important factor to assure the bioavailability of plant extracts intended to be administered via the oral route. The aim of this work was to study the stability of a lyophilized aqueous extract of L. divaricata compressed as a pill. To this end, the main polyphenol compound found in the extract, that is, the nordihydroguaiaretic acid, the total polyphenols and flavonoids content and the antioxidant activity such as diphenylpicrylhydrazyl scavenger activity and reducing power were assayed after subjecting the extract to different incubation times in simulated digestive fluids. The HPLC and spectroscopic methods were employed. Although the levels of polyphenols and flavonoids decreased upon incubation in gastric and intestinal fluids, the extract maintained its antioxidant activity related to the presence of nordihydroguaiaretic acid. These results are promising and encourage the potential use of the extract by the oral route as a supplement or phytomedicine with antioxidant activity. Copyright © 2017 John Wiley & Sons, Ltd.

Polyphenol-Rich Fraction from Larrea divaricata and its Main Flavonoid Quercetin-3-Methyl Ether Induce Apoptosis in Lymphoma Cells Through Nitrosative Stress.

Larrea divaricata is a plant with antiproliferative principles. We have previously identified the flavonoid quercetin-3-methyl ether (Q-3-ME) in an ethyl acetate fraction (EA). Both the extract and Q-3-ME were found to be effective against the EL-4 T lymphoma cell line. However, the mechanism underlying the inhibition of tumor cell proliferation remains to be elucidated. In this work, we analyzed the role of nitric oxide (NO) in the induction of apoptosis mediated by Q-3-ME and EA. Both treatments were able to induce apoptosis in a concentration-dependent and time-dependent manner. The western blot analysis revealed a sequential activation of caspases-9 and 3, followed by poly-(ADP-ribose)-polymerase cleavage. EA and Q-3-ME lowered the mitochondrial membrane potential, showing the activation of the intrinsic pathway of apoptosis. Q-3-ME and EA increased NO production and inducible NO synthase expression in tumor cells. The involvement of NO in cell death was confirmed by the nitric oxide synthases inhibitor L-NAME. In addition, EA and Q-3-ME induced a cell cycle arrest in G0/G1 phase. These drugs did not affect normal cell viability. This data suggested that EA and Q-3-ME induce an increase in NO production that would lead to the cell cycle arrest and the activation of the intrinsic pathway of apoptosis. Copyright © 2016 John Wiley & Sons, Ltd.

A fraction containing kaempferol-3,4'-dimethylether from Larrea divaricata Cav. induces macrophage activation on mice infected with Candida albicans.

Larrea divaricata Cav. is a plant growing in South America. Both the infusion and a derived fraction (F1) of L. divaricata have proved to have immunomodulatory properties. Moreover, F1 can activate macrophages obtained from mice infected with Candida albicans. In this work, F1 was administrated to infected animals, and the state and type of activation of resident macrophages were studied. Results showed that F1 was able to activate macrophages obtained from infected mice by both classical and alternative pathways, probably by inducing a translocation of nuclear factor kappa-B. F1 increases not only the lysosomal activity of macrophages but also the production of phagosomal superoxide anion as a consequence of the activation of the Nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase) complex. F1 induced an increase in the macrophage capacity to kill the fungus, which was reflected in a decrease in the levels of colonization of organs. A main flavonoid, kaempferol-3,4'-dimethylether, was identified in F1 by HPLC. This compound increased in vitro production of nitric oxide in heat-killed C. albicans-stimulated macrophages. The flavonoid could thus be considered one of the responsible molecules mediating the overall effects of F1 on the immune system in infected animals.

A fraction rich in phenyl propanoids from L. divaricata aqueous extract is capable of inducing apoptosis, in relation to H2O2 modulation, on a murine lymphoma cell line.

Leukemia and lymphoma are a group of heterogeneous neoplastic disorder of white blood cells characterized by the uncontrolled proliferation and block in differentiation of hematopoietic cells. Nowadays, there is an interest in therapy with drugs of plant origin because conventional medicine can be inefficient or also results in side effects. Larrea divaricata Cav., is a plant widely distributed in Argentina that possess antiproliferative and antioxidant activities reported. Nordihydroguaiaretic acid (NDGA) was previously found in the plant and related to both antiproliferative and pro-proliferative actions on a lymphoma cell line. In order to demonstrate whether the presence of NDGA may be beneficial or not in the antiproliferative action of the aqueous extract, the extract of L. divaricata was submitted to a fractionation and fractions with and without NDGA were studied in a murine lymphocitic leukemia cell line (EL-4) proliferation. The effect of the most active fraction was studied in relation to H2O2 modulation and the synergistic action between compounds, found in fractions, was analyzed. The presence of NDGA was not a detonator for pro-proliferative action and its presence could be beneficial in low concentrations allowing a synergist antiproliferative action with other compounds.

Larrea divaricata Cav: scientific evidence showing its beneficial effects and its wide potential application / Larrea divaricata Cav: evidencia específica mostrando sus efectos beneficiosos y su amplia aplicación potencial

Larrea divaricata Cav. (jarilla) (Zygophyllaceae) is used in Argentinean folk medicine. It contains nordihydroguaiaretic acid (NDGA), a lignan with nefrotoxic and hepatotoxic effects. The presence of NDGA converts controversial the use of L. divaricata. The amount of NDGA is higher in alcoholic extracts than in aqueous extracts (AE). The last 20 years had a great advance on the use of AE to treat different conditions in a safe manner. In this review, we present the scientific results tending to confirm the potential beneficial effects of the AE on human health. The anti-proliferative effects of AE of L. divaricata have been assayed on a tumor lymphoid line (BW 5147) and the pathways involved in such effects were described. The anti-microbial activity was determined by tests for bacteria and fungus. The anti-inflammatory activity was assayed by using carrageen and TPA induced-inflammation tests. The Immunomodulatory effects were investigated ¯in vivo and ¯in vitro on mice. Sub-fractions of aqueous extracts were obtained and analyzed. The immunogenicity of proteins from crude AE was characterized and antioxidant and nutritional activity were studied. The effect of an AE on hair loss was assayed. In summary, AE from L. divaricata has pharmacological activities including anti-microbial, anti-inflammatory, anti-cancer effects.

Larrea divaricata Cav. (Jarilla) (Zygophyllaceae) es utilizada en medicina popular Argentina. Contiene ácido nordihidroguaiarético (NDGA), un lignano con efectos nefrotóxicos y hepatotóxicos. La presencia de NDGA hace controvertido el uso de L. divaricata. La cantidad de NDGA es mayor en extractos alcohólicos que en extractos acuosos (EA). Los últimos 20 años han tenido un gran avance en el uso de EA para el tratamiento de diferentes condiciones en forma segura. En esta revisión se presentan resultados científicos que confirman los efectos potencialmente beneficiosos de los EA sobre la salud humana. Los efectos anti-proliferativos se han ensayado en una línea de tumor linfoide, así como los mecanismos involucrados. La actividad anti-microbiana se determinó usando pruebas para bacterias y hongos. La actividad anti-inflamatoria fue evaluada mediante el uso de inflamación inducida por carragenina y TPA. Los efectos inmunomoduladores fueron investigados "in vivo" e "in vitro" en ratones. Sub-fracciones de los EA fueron obtenidos y analizados. La inmunogenicidad de las proteínas del EA crudo se caracterizaron y se estudiaron las actividades antioxidantes nutricionales. Además, se ensayó el efecto del EA en la caída del cabello. En resumen, los EA de L. divaricata presentan actividades farmacológicas como por ejemplo, anti-microbiana, anti-inflamatoria y efectos anti-cancerígenos.

Dermis acelular porcina (sus scrofa) cargada con antioxidantes de larrea divaricata / Porcine (sus scrofa) acellular dermis load with antioxidants from larrea divaricata

Con el objeto de cargar con antioxidantes de Larrea divaricata una dermis acelular porcina para propósitos terapéuticos, se determinó el contenido de polifenoles y antocianinas de extractos puros, aislados y absorbidos en una dermis acelular porcina. Los valores para polifenoles totales y antocianinas fueron: a) Larrea divaricata: 58,77 + 1,55 mg ácido gálico / 100 g peso fresco, 400,00 + 9,55 mg cianidina 3-glucósido / 100 g peso fresco, repectivamente, b) dermis acelular porcina: 8,86 + 0,55 mg ac. gállico / 100 g peso fresco y 0,10+ 0,00 mg cianidina 3-glucósido / 100 g peso fresco; respectivamente, c) Larrea divaricata absorbida en dermis acelular porcina 45,92 + 0,90 mg ácido gálico / 100 g peso fresco y 155,92 + 5,90 mg cianidina 3-glucósido / 100 g peso fresco, respectivamente. Nosotros concluimos que es posible tener una dermis acelular porcina cargada con antioxidantes de Larrea divaricata para propósitos médicos.

The aim of the study was to evaluate loading with antioxidants from Larrea divaricata a porcine acellular dermis for therapeutic purposes, poliphenols and anthocianins of pure extracts, isolated and absorbed in pig acellular dermis was evaluated. The following values (total polyphenols and anthocianins) were obtained: a) Larrea divaricata: 58,77 + 1,55 mg gallic acid / 100 g fresh weight; 400,95 + 9,55 mg cianydin 3- glucosyde / 100 g fresh weight; respectively; b) porcine acellular dermis: 8,86 + 0,55 mg gallic acid / 100 g fresh weight and 0,10+ 0,00 mg cianydin 3-glucosyde / 100 g fresh weight; respectively, c) L. divaricata absorbed in porcine acellular dermis: 45,92 + 0,90 mg gallic acid / 100 g fresh weight and 155,92 + 5,90 mg cianydin 3-glucosyde / 100 g fresh weight, respectively. We concluded that it is possible to get a porcine acellular dermis loaded with antioxidants from Larrea divaricata for medical purposes.