Matteucinol combined with temozolomide inhibits glioblastoma proliferation, invasion, and progression: an in vitro, in silico, and in vivo study

Glioblastoma is the most prevalent and malignant brain tumor identified in adults. Surgical resection followed by radiotherapy and chemotherapy, mainly with temozolomide (TMZ), is the chosen treatment for this type of tumor. However, the average survival of patients is around 15 months. Novel approaches to glioblastoma treatment are greatly needed. Here, we aimed to investigate the anti-glioblastoma effect of the combination of matteucinol (Mat) (dihydroxyflavanone derived from Miconia chamissois Naudin) with the chemotherapeutic TMZ in vitro using tumor (U-251MG) and normal astrocyte (NHA) cell lines and in vivo using the chick embryo chorioallantoic membrane (CAM) assay. The combination was cytotoxic and selective for tumor cells (28 μg/mL Mat and 9.71 μg/mL TMZ). Additionally, the combination did not alter cell adhesion but caused morphological changes characteristic of apoptosis in vitro. Notably, the combination was also able to reduce tumor growth in the chick embryo model (CAM assay). The docking results showed that Mat was the best ligand to the cell death membrane receptor TNFR1 and to TNFR1/TMZ complex, suggesting that these two molecules may be working together increasing their potential. In conclusion, Mat-TMZ can be a good candidate for pharmacokinetic studies in view of clinical use for the treatment of glioblastoma.

In vivo effect of essential oil of Mentha x villosa and its active compound against Schistosoma mansoni (Sambon, 1907) / Efeito in vivo do óleo essencial de Mentha x villosa e seu composto ativo sobre Schistosoma mansoni (Sambon, 1907)

Abstract Schistosomiasis treatment is dependent on a single drug, praziquantel (PZQ). The development of resistance of PZQ has drawn the attention of many researchers to alternative drugs. One viable and promising treatment is the study of medicinal plants as a new approach to the experimental treatment for Schistosomiasis. The present work aimed to evaluate in vivo antischistosomal activity of effect of Mentha x villosa Oil Essential (Mv-EO) and rotundifolone (ROT) against Schistosoma mansoni. Thirty-day-old female Swiss webster mice (Mus musculus) weighing 50 grams were used. Mice were infected with 80 cercariae of S. mansoni (BH strain) and orally administered Mv-EO (50, 100 and 200 mg/Kg) and ROT (35.9, 70.9 and 141.9 mg/Kg) at 45-days post infection for 5 consecutive days. All mice were euthanized 60 days after infection. Praziquantel was the positive control in the experiment. Doses of 200 mg/kg (Mv-EO) and ROT (141.9 mg/Kg) resulted in a significant reduction in fluke burden (72.44% and 74.48%, respectively). There was also marked reduction in liver, intestinal and faecal and changed oogram pattern, compared to infected untreated mice. Considering the results obtained, further biological studies are required in order to elucidate the mechanism of schistosomicidal action on against adult S. mansoni.

Resumo O tratamento da esquistossomose é dependente de uma única droga, praziquantel (PZQ). O desenvolvimento da resistência de PZQ tem atraído atenção de muitos pesquisadores por medicamentos alternativos. Um tratamento viável e promissor é o estudo das plantas medicinais como uma nova abordagem para o tratamento experimental para esquistossomose. O presente trabalho objetivou avaliar a atividade esquistossomicida in vivo óleo essencial de Mentha x villosa (OE-Mv) e rotundifolona (ROT) contra Schistosoma mansoni. Foram utilizados camundongos Swiss webster (Mus musculus) fêmea de trinta dias de idade pesando 50 gramas. Os camundongos foram infectados com 80 cercárias de S. mansoni (cepa BH) e administrado por via oral OE-Mv (50, 100 e 200 mg/Kg) e ROT (35,9, 70,9 e 141,9 mg/Kg) apos 45 dias de infecção durante 5 dias consecutivos. Todos os animais foram eutanasiados 60 dias após a infecção. Praziquantel foi o controle positivo no experimento. O tratamento dos camundongos infectados com doses de 200 mg/kg (OE-Mv) e rotundifolona (141,9 mg/Kg) resultaram em redução significativa dos vermes (72.44% e 74.48%, respectivamente). Foi observado também redução no fígado, intestino e fecal e alteração no padrão do oograma, em comparação aos camundongos infectados e não tratados. Considerando os resultados obtidos, mais estudos biológicos são necessários a fim de elucidar o mecanismo de ação esquistossomicida contra adultos de S. mansoni.

In vivo effect of essential oil of Mentha x villosa and its active compound against Schistosoma mansoni (Sambon, 1907)

Abstract Schistosomiasis treatment is dependent on a single drug, praziquantel (PZQ). The development of resistance of PZQ has drawn the attention of many researchers to alternative drugs. One viable and promising treatment is the study of medicinal plants as a new approach to the experimental treatment for Schistosomiasis. The present work aimed to evaluate in vivo antischistosomal activity of effect of Mentha x villosa Oil Essential (Mv-EO) and rotundifolone (ROT) against Schistosoma mansoni. Thirty-day-old female Swiss webster mice (Mus musculus) weighing 50 grams were used. Mice were infected with 80 cercariae of S. mansoni (BH strain) and orally administered Mv-EO (50, 100 and 200 mg/Kg) and ROT (35.9, 70.9 and 141.9 mg/Kg) at 45-days post infection for 5 consecutive days. All mice were euthanized 60 days after infection. Praziquantel was the positive control in the experiment. Doses of 200 mg/kg (Mv-EO) and ROT (141.9 mg/Kg) resulted in a significant reduction in fluke burden (72.44% and 74.48%, respectively). There was also marked reduction in liver, intestinal and faecal and changed oogram pattern, compared to infected untreated mice. Considering the results obtained, further biological studies are required in order to elucidate the mechanism of schistosomicidal action on against adult S. mansoni.

Resumo O tratamento da esquistossomose é dependente de uma única droga, praziquantel (PZQ). O desenvolvimento da resistência de PZQ tem atraído atenção de muitos pesquisadores por medicamentos alternativos. Um tratamento viável e promissor é o estudo das plantas medicinais como uma nova abordagem para o tratamento experimental para esquistossomose. O presente trabalho objetivou avaliar a atividade esquistossomicida in vivo óleo essencial de Mentha x villosa (OE-Mv) e rotundifolona (ROT) contra Schistosoma mansoni. Foram utilizados camundongos Swiss webster (Mus musculus) fêmea de trinta dias de idade pesando 50 gramas. Os camundongos foram infectados com 80 cercárias de S. mansoni (cepa BH) e administrado por via oral OE-Mv (50, 100 e 200 mg/Kg) e ROT (35,9, 70,9 e 141,9 mg/Kg) apos 45 dias de infecção durante 5 dias consecutivos. Todos os animais foram eutanasiados 60 dias após a infecção. Praziquantel foi o controle positivo no experimento. O tratamento dos camundongos infectados com doses de 200 mg/kg (OE-Mv) e rotundifolona (141,9 mg/Kg) resultaram em redução significativa dos vermes (72.44% e 74.48%, respectivamente). Foi observado também redução no fígado, intestino e fecal e alteração no padrão do oograma, em comparação aos camundongos infectados e não tratados. Considerando os resultados obtidos, mais estudos biológicos são necessários a fim de elucidar o mecanismo de ação esquistossomicida contra adultos de S. mansoni.

Immune cells and oxidative stress in the endotoxin tolerance mouse model

Sepsis is a systemic inflammatory response that can lead to tissue damage and death. In order to increase our understanding of sepsis, experimental models are needed that produce relevant immune and inflammatory responses during a septic event. We describe a lipopolysaccharide tolerance mouse model to characterize the cellular and molecular alterations of immune cells during sepsis. The model presents a typical lipopolysaccharide tolerance pattern in which tolerance is related to decreased production and secretion of cytokines after a subsequent exposure to a lethal dose of lipopolysaccharide. The initial lipopolysaccharide exposure also altered the expression patterns of cytokines and was followed by an 8- and a 1.5-fold increase in the T helper 1 and 2 cell subpopulations. Behavioral data indicate a decrease in spontaneous activity and an increase in body temperature following exposure to lipopolysaccharide. In contrast, tolerant animals maintained production of reactive oxygen species and nitric oxide when terminally challenged by cecal ligation and puncture (CLP). Survival study after CLP showed protection in tolerant compared to naive animals. Spleen mass increased in tolerant animals followed by increases of B lymphocytes and subpopulation Th1 cells. An increase in the number of stem cells was found in spleen and bone marrow. We also showed that administration of spleen or bone marrow cells from tolerant to naive animals transfers the acquired resistance status. In conclusion, lipopolysaccharide tolerance is a natural reprogramming of the immune system that increases the number of immune cells, particularly T helper 1 cells, and does not reduce oxidative stress.

Cloning of a novel acidic phospholipase A2 from the venom gland of Crotalus durissus cascavella (Brazilian northeastern rattlesnake)

The phospholipase A2 superfamily encompasses 15 groups that are classified into: secreted PLA2 (sPLA2); cytosolic PLA2 (cPLA2); Ca2+-independent intracellular PLA2 (iPLA2); platelet-activating factor acetylhydrolase (PAF-AH); and lysosomal PLA2. Currently, approximately 700 PLA2 sequences are known, of which 200 are obtained from the venom gland of Crotalinae snakes. However, thus far, little information is available on cloning, purification and structural characterization of PLA2 from Crotalus durisssus cascavela venom gland. In the present work, we report the molecular cloning of a novel svPLA2 from C. d. cascavella (Cdc), a predominant rattlesnake subspecies in northeastern Brazil. The Cdc svPLA2 cDNA precursor is 689 nucleotides long and encodes a protein of 138 amino acid residues, with a calculated molecular mass of approximately 13,847 Da and an estimated isoelectric point of 5.14. Phylogenetic analysis of Crotalinae PLA2 reveals that Cdc PLA2 clustered with other acidic type IIA PLA2 homologues is also present in the venom of North American rattlesnakes. Hitherto, this study presents a novel PLA2 cDNA precursor from C. d. cascavella and data reported herein will be useful for further steps in svPLA2 purification and analysis.