Utilization of the Plant Clusia Fluminensis Planch & Triana Against Some Toxic Activities of the Venom of Bothrops jararaca and B. jararacussu Snake Venom Toxic Activities.

BACKGROUND: In Brazil, the Bothrops genus accounts for 87% of registered snakebites, which are characterized by hemorrhage, tissue necrosis, hemostatic disturbances, and death. The treatment recommended by governments is the administration of specific antivenoms. Although antivenom efficiently prevents venom-induced lethality, it has limited efficacy in terms of preventing local tissue damage. Thus, researchers are seeking alternative therapies able to inhibit the main toxic effects of venoms, without compromising safety. OBJECTIVE: The study aimed to test the ability of aqueous extracts of leaves, stems, and fruits of the plant Clusia fluminensis to neutralize some toxic effects induced by the venoms of Bothrops jararaca and Bothrops jararacussu. METHODS: The plant extracts were incubated with venoms for 30 min. at 25 °C, and then in vitro (coagulant and proteolytic) and in vivo (hemorrhagic, myotoxic, and edematogenic) activities were evaluated. In addition, the extracts were administered to animals (by oral, intravenous or subcutaneous routes) before or after the injection of venom samples, and then hemorrhage and edema assays were performed. In addition, a gel solution of the fruit extract was produced and tested in terms of reducing hemorrhage effects. A chemical prospection was performed to identify the main classes of compounds present in the extracts. RESULTS: All the extracts inhibited the activities of the two venoms, regardless of the experimental protocol or route of administration of the extracts. Moreover, the gel of the fruit extract inhibited the venom-induced-hemorrhage. The extracts comprised of tannins, flavonoids, saponins, steroids, and terpenoids. CONCLUSION: Antivenom properties of C. fluminensis extracts deserve further investigation in order to gain detailed knowledge regarding the neutralization profile of these extracts.

Clusia criuva Cambess. (Clusiaceae): anatomical characterization, chemical prospecting and antioxidant activity

ABSTRACT This study aims the anatomical description and chemical characterization of aerial parts of Clusia criuva Cambess., Clusiaceae in addition to the evaluation of the antioxidant activity of crude extracts, correlated to the flavonoid content. The morphological characterization was performed using traditional techniques of plant anatomy. For phytochemical studies, crude extracts were obtained by static maceration and analyzed by thin layer chromatography. The antioxidant activity and the flavonoids content were determined by colorimetric methods involving, respectively, 2,2-diphenyl-1-picrylhydrazyl free radical and aluminum chloride. C. criuva has uniseriate epidermis, paracytic stomata, hypostomatic leaves, cuticular flanges and cordiform vascular cylinder with accessory bundles. Chemical prospecting confirmed the abundant presence of terpenes and phenols in the extracts of leaves and of fruits. The methanolic extract of seeds showed the lowest EC50 value, but the methanolic extract of pericarps exhibited the highest maximum antioxidant activity. The results suggested a high percentage of flavonoids in the hexanic extract of pericarps, however, this could represent, in fact, the presence of benzophenones. Secretory ducts and the shape of the midrib are diagnostic for C. criuva. The antioxidant activity is not directly related to the flavonoids. The results indicate the importance of future studies with C. criuva chemical constituents.

Clusia criuva Cambess. (Clusiaceae): anatomical characterization, chemical prospecting and antioxidant activity.

This study aims the anatomical description and chemical characterization of aerial parts of Clusia criuva Cambess., Clusiaceae in addition to the evaluation of the antioxidant activity of crude extracts, correlated to the flavonoid content. The morphological characterization was performed using traditional techniques of plant anatomy. For phytochemical studies, crude extracts were obtained by static maceration and analyzed by thin layer chromatography. The antioxidant activity and the flavonoids content were determined by colorimetric methods involving, respectively, 2,2-diphenyl-1-picrylhydrazyl free radical and aluminum chloride. C. criuva has uniseriate epidermis, paracytic stomata, hypostomatic leaves, cuticular flanges and cordiform vascular cylinder with accessory bundles. Chemical prospecting confirmed the abundant presence of terpenes and phenols in the extracts of leaves and of fruits. The methanolic extract of seeds showed the lowest EC50 value, but the methanolic extract of pericarps exhibited the highest maximum antioxidant activity. The results suggested a high percentage of flavonoids in the hexanic extract of pericarps, however, this could represent, in fact, the presence of benzophenones. Secretory ducts and the shape of the midrib are diagnostic for C. criuva. The antioxidant activity is not directly related to the flavonoids. The results indicate the importance of future studies with C. criuva chemical constituents.

Chemosystematic aspects of polyisoprenylated benzophenones from the genus Clusia.

Benzophenone derivatives are special metabolites that arouse great scientific interest. The Clusiaceae family is known for producing large amounts of benzophenone derivatives with several isoprene residues on their structures, which are responsible for the observed complexity and structural variety in this class of substances, and also contribute to their biological activities. Clusia is an important genus belonging to Clusiaceae, with 55 different polyisoprenylated benzophenones identified so far. These substances were analyzed from biosynthetic and chemosystematic points of view, allowing the determination of characteristics regarding their production, accumulation and distribution within this genus. Polyisoprenylated benzophenones found in Clusia showed a high prenylation degree, with 2 to 5 isoprene units and a greater occurrence in flowers and fruits. Section Cordylandra showed a very similar occurrence of 2,4,6-trihydroxybenzophenone derivatives and bicyclo[3.3.1]nonane-2,4,9-trione derivatives, the majority of them with 4 isoprene units. In section Anandrogyne there is a predominance of simple 2,4,6-trihydroxy-benzophenone derivatives, with 2 isoprene units, and in Chlamydoclusia predominates bicyclo[3.3.1]nonane-2,4,9-trione derivatives with 4 isoprene units. Although highly prenylated, these substances showed low oxidation indexes, which from an evolutionary perspective corroborates the fact that Clusiaceae is a family in transition, with some common aspects with both basal and derived botanical families.

Cytotoxic and Antiviral Activity of Extracts and Compounds Isolated from Clusia Fluminensis Planch. & Triana (Clusiaceae) / Atividade Citotóxica e Antiviral de Extratos e Compostos Isolados de Clusia Fluminensis Planch. & Triana (Clusiaceae)

The worldwide distribution of herpes simplex virus type 1 (HSV-1) allied to the emergence of resistant strains makes necessary and urgent the search and development of new substances capable of preventing and treating HSV-1 infections. Studies demonstrate synergy between genital herpes and human immunodeficiency virus type 1 (HIV-1), which represents a major concern for global public health. Objective: The objective of this study was to evaluate the activity of crude extracts and isolated substances from C. fluminensis in the in vitro replication of the HSV-1 virus and HIV-1-RT activity. Methods: This study evaluated the activity of extracts and isolated compounds from Clusia fluminensis Planch. & Triana against HSV-1 using Vero cells in culture and against HIV-1 using a recombinant reverse transcriptase enzyme (HIV -1 RT). The percentage of inhibition against HSV-1 was determined from viral lysis plaque reduction assay, and the anti-HIV-1-RT test was performed by a fluorimetric assay. It was also evaluated the cytotoxic activity of the samples using MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]. Results: The crude extracts showed high percentage of inhibition against HSV-1, reaching 81.4 to 100.0% inhibition in non-cytotoxic concentration (50 µg/mL). The isolated compounds, lanosterol and clusianone, demonstrated 100% inhibition in non-cytotoxic concentration (50 µg/mL). We also examined the effects of the extracts and isolates on the activity of the HIV-1-RT. Among the crude extracts, only the methanolic extract of leaves and methanolic extract of stems showed inhibitory activity against HIV-1-RT. Regarding the isolated compounds, lanosterol showed a moderate activity. Conclusion: Our data demonstrate that extracts and isolates compounds Clusia fluminensis Planch. & Triana have promising antiviral activity inhibiting HSV-1 replication and HIV-1 by inhibiting the anti-RT activity.

A distribuição mundial do vírus herpes simplex tipo 1 (HSV-1) aliada ao surgimento de cepas resistentes torna necessária e urgente a busca e o desenvolvimento de novas substâncias capazes de prevenir e tratar infecções HSV-1. Estudos demonstram sinergia entre herpes genital e vírus da imunodeficiência humana tipo 1 (HIV-1), o que representa uma grande preocupação para a saúde pública global. Objetivo: O objetivo deste estudo foi avaliar a atividade de extratos brutos e substâncias isoladas de Clusia fluminensis Planch. & Triana na replicação in vitro do vírus HSV-1 e na atividade anti HIV-1-RT. Métodos: Este estudo avaliou a atividade de extratos e substâncias isoladas de Clusia fluminensis Planch. & Triana contra o HSV-1 utilizando células Vero em cultura e contra o HIV-1 utilizando a enzima transcriptase reversa recombinante (HIV-1 RT). A porcentagem de inibição contra o HSV-1 foi determinada a partir do ensaio de redução de placas de lise viral, e o ensaio anti-HIV-1 RT foi realizado por um ensaio fluorimétrico. Também foi avaliada a atividade citotóxica das amostras utilizando MTT [brometo de 3- (4,5-dimetiltiazol-2-il) -2,5-difeniltetrazólio]. Resultados: Os extratos demonstraram elevada percentagem de inibição contra o HSV-1, atingindo 81,4 a 100,0% de inibição em concentração não citotóxica (50 µg/mL). Os compostos isolados, lanosterol e clusianona, demonstraram 100% de inibição em concentração não citotóxica (50 µg/mL). Examinamos também os efeitos dos extratos e isolados sobre a atividade anti-HIV-1 RT. Entre os extratos brutos, apenas o extrato metanólico das folhas e caules apresentaram atividade anti-HIV-1 RT. Em relação aos compostos isolados, lanosterol mostrou uma atividade moderada. Conclusão: Nossos dados demonstram que os extratos e compostos isolados de Clusia fluminensis Planch. & Triana possuem atividade antiviral promissora inibindo a replicação do HSV-1 e HIV-1 através da inibição da atividade anti-RT.

Inhibitory effect of the plant Clusia fluminensis against biological activities of Bothrops jararaca snake venom.

The ability of extracts of the plant Clusia fluminensis Planch & Triana (Clusiaceae Lindl.) to neutralize proteolysis, clotting, hemolysis, hemorrhagic and lethality activities of Bothrops jararaca snake venom was studied. Clusianone and lanosterol from the flower and fruit extracts, respectively, were also tested. The extracts of different organs of C. fluminensis inhibited proteolysis and hemolysis induced by B. jararaca venom, but with different potencies. Only the stems prevented blood clotting. Only the acetone extract of the fruit protected mice from hemorrhage while the acetone or methanol extracts prevented mice from death. Clusianone and lanosterol did not inhibit clotting or hemorrhage, but the former inhibited proteolysis and the latter hemolysis.