Enhancement of Annexin V in response to combination of epigallocatechin gallate and quercetin as a potent arrest the cell cycle of colorectal cancer / Aumento da anexina V em resposta à combinação de galato de epigalocatequina e quercetina como uma potente parada do ciclo celular do câncer colorretal

Colorectal cancer (CRC) is one of the most common cancers leading to comorbidities and mortalities globally. The rational of current study was to evaluate the combined epigallocatechin gallate and quercetin as a potent antitumor agent as commentary agent for therapeutic protocol. The present study investigated the effect of epigallocatechin Gallate (EGCG) (150mg) and quercetin (200mg) at different proportions on proliferation and induction of apoptosis in human colon cancer cells (HCT-116). Cell growth, colonogenic, Annexin V in addition cell cycle were detected in response to phytomolecules. Data obtained showed that, the colony formation was inhibited significantly in CRC starting from the lowest concentration tested of 10 µg/mL resulting in no colonies as visualized by a phase-contrast microscope. Data showed a significant elevation in the annexin V at 100 µg/mL EGCG(25.85%) and 150 µg/mL quercetin (48.35%). Moreover, cell cycle analysis showed that this combination caused cell cycle arrest at the G1 phase at concentration of 100 µg/mL (72.7%) and 150 µg/mL (75.25%). The combined effect of epigallocatechin Gallate and quercetin exert antiproliferative activity against CRC, it is promising in alternative conventional chemotherapeutic agent.

O câncer colorretal (CCR) é um dos cânceres mais comuns, levando a comorbidades e mortalidade em todo o mundo. O racional do presente estudo foi avaliar a combinação de galato de epigalocatequina e quercetina como um agente antitumoral potente como agente de comentário para protocolo terapêutico. O presente estudo investigou o efeito de galato de epigalocatequina (EGCG) (150 mg) e quercetina (200 mg) em diferentes proporções na proliferação e indução de apoptose em células de câncer de cólon humano (HCT-116). O crescimento celular, colonogênico, anexina V, além do ciclo celular foram detectados em resposta a fitomoléculas. Os dados obtidos mostraram que a formação de colônias foi inibida significativamente no CRC a partir da concentração mais baixa testada de 10 µg/mL, resultando em nenhuma colônia conforme visualizado por um microscópio de contraste de fase. Os dados mostraram uma elevação significativa na anexina V a 100 µg/mL de EGCG (25,85%) e 150 µg/mL de quercetina (48,35%). Além disso, a análise do ciclo celular mostrou que essa combinação causou parada do ciclo celular na fase G1 na concentração de 100 µg/mL (72,7%) e 150 µg/mL (75,25%). O efeito combinado da epigalocatequina galato e quercetina exerce atividade antiproliferativa contra o CCR, é promissor como agente quimioterápico alternativo convencional.

Enhancement of Annexin V in response to combination of epigallocatechin gallate and quercetin as a potent arrest the cell cycle of colorectal cancer / Aumento da anexina V em resposta à combinação de galato de epigalocatequina e quercetina como uma potente parada do ciclo celular do câncer colorretal

Colorectal cancer (CRC) is one of the most common cancers leading to comorbidities and mortalities globally. The rational of current study was to evaluate the combined epigallocatechin gallate and quercetin as a potent antitumor agent as commentary agent for therapeutic protocol. The present study investigated the effect of epigallocatechin Gallate (EGCG) (150mg) and quercetin (200mg) at different proportions on proliferation and induction of apoptosis in human colon cancer cells (HCT-116). Cell growth, colonogenic, Annexin V in addition cell cycle were detected in response to phytomolecules. Data obtained showed that, the colony formation was inhibited significantly in CRC starting from the lowest concentration tested of 10 µg/mL resulting in no colonies as visualized by a phase-contrast microscope. Data showed a significant elevation in the annexin V at 100 µg/mL EGCG(25.85%) and 150 µg/mL quercetin (48.35%). Moreover, cell cycle analysis showed that this combination caused cell cycle arrest at the G1 phase at concentration of 100 µg/mL (72.7%) and 150 µg/mL (75.25%). The combined effect of epigallocatechin Gallate and quercetin exert antiproliferative activity against CRC, it is promising in alternative conventional chemotherapeutic agent.(AU)

O câncer colorretal (CCR) é um dos cânceres mais comuns, levando a comorbidades e mortalidade em todo o mundo. O racional do presente estudo foi avaliar a combinação de galato de epigalocatequina e quercetina como um agente antitumoral potente como agente de comentário para protocolo terapêutico. O presente estudo investigou o efeito de galato de epigalocatequina (EGCG) (150 mg) e quercetina (200 mg) em diferentes proporções na proliferação e indução de apoptose em células de câncer de cólon humano (HCT-116). O crescimento celular, colonogênico, anexina V, além do ciclo celular foram detectados em resposta a fitomoléculas. Os dados obtidos mostraram que a formação de colônias foi inibida significativamente no CRC a partir da concentração mais baixa testada de 10 µg/mL, resultando em nenhuma colônia conforme visualizado por um microscópio de contraste de fase. Os dados mostraram uma elevação significativa na anexina V a 100 µg/mL de EGCG (25,85%) e 150 µg/mL de quercetina (48,35%). Além disso, a análise do ciclo celular mostrou que essa combinação causou parada do ciclo celular na fase G1 na concentração de 100 µg/mL (72,7%) e 150 µg/mL (75,25%). O efeito combinado da epigalocatequina galato e quercetina exerce atividade antiproliferativa contra o CCR, é promissor como agente quimioterápico alternativo convencional.(AU)

Quercetin associated with dimethylsulfoxide has a curative effect on experimental colon anastomosis injury

Purpose To examine the effects of quercetin on healing of experimental colon anastomosis injury in early and late period. Methods Eighty male Wistar-Albino rats were divided into 8 groups. For all groups, left colons of the rats were resected and for the rest end-to-end anastomosis was performed. Two of the groups for which the experiment protocol was ended on the 3rd and 7th day following the anastomosis were not administered with either quercetin or dimethylsulfoxide DMSO, whereas two other groups were administered with DMSO only, and four other groups were administered with quercetin dissolved in DMSO in doses of 20 and 100 mg/kg during the protocol. At the end of the study, anastomosis line was resected, histopathological evaluation was performed and bursting pressure, malondialdehyde, superoxide dismutase, catalase, and hydroxyproline levels were measured. Results Quercetin significantly increased hydroxyproline, superoxide dismutase, catalase levels, histopathological healing score, bursting pressure values and decreased malondialdehyde level in early period. It also significantly increased superoxide dismutase, catalase, and hydroxyproline levels and decreased malondialdehyde level in late period. Conclusion It was seen that quercetin speeds up the injury healing process and reveals an antioxidant effect, specifically in early period.(AU)

Uso de quercetina em meio de refrigeração de sêmen equino a base de leite em pó desnatado / Use of quercetin in equine semen cooling medium based on skimded milk poder

A biotécnica de refrigeração de sêmen equino para o trasporte de material genético está amplamente distribuida. Com a necessidade da manutenção da viabilidade espermática é indispensável a autilização e diluentes para este fim. O objetivo deste estudo foi avaliar se a adição de quercetina, que é um flavonóide antioxidante, em diluentes espermáticos altera a viabilidade dos espermatozoides equinos refrigerados. Foram avaliados cinco ejaculados de um garanhão, por meio da análise de motilidade e vigor espermático, em diferentes períodos de refrigeração em quatro meios de diluição: BotuSêmen®; BotuSêmen® adicionado de 20 µg/mL de quercetina (SIGMA®); Solução aquosa com 10% leite em pó desnatado (Molico®) ou Solução aquosa com 10% leite em pó desnatado (Molico®) adicionado de 20 µg/mL de quercetina (SIGMA®). Por meio dos resultados obtidos no presente estudo, foi possível verificar que não há diferença estatística entre a motilidade espermática de sêmen de garanhão diluído em BotuSêmen® em relação ao diluído solução aquosa com 10% leite em pó desnatado (Molico®) pelo período de refrigeração avaliado (36h).

The biotechnique of equine semen cooling for the transfer of genetic material is widely distributed. With the need to maintain sperm viability, it is indispensable to use and thinners for this purpose. The aim of this study was to evaluate whether the addition of quercetin, which is an anti-oxidant flavonoid, in spermatic diluents alters the viability of refrigerated equine spermatozoa. Five ejaculates of a stallion were evaluated by analysis of motility and spermatic vigor in different cooling periods in four dilution media: BotuSêmen® ; BotuSêmen® added 20 µg/mL of quercetin (SIGMA® ); Aqueous solution with 10% skimmilk powder (Molico® ) or Aqueous solution with 10% skimmilk powder (Molico® ) added 20 µg/mL quercetin (SIGMA® ). Through the data obtained, it was possible to verify that there is no statistical difference between the sperm motility of diluted stallion semen in BotuSêmen® in relation to the diluted aqueous solution with 10% skimmed milk powder (Molico® ) for the evaluated refrigeration period (36h).

Uso de quercetina em meio de refrigeração de sêmen equino a base de leite em pó desnatado / Use of quercetin in equine semen cooling medium based on skimded milk poder

A biotécnica de refrigeração de sêmen equino para o trasporte de material genético está amplamente distribuida. Com a necessidade da manutenção da viabilidade espermática é indispensável a autilização e diluentes para este fim. O objetivo deste estudo foi avaliar se a adição de quercetina, que é um flavonóide antioxidante, em diluentes espermáticos altera a viabilidade dos espermatozoides equinos refrigerados. Foram avaliados cinco ejaculados de um garanhão, por meio da análise de motilidade e vigor espermático, em diferentes períodos de refrigeração em quatro meios de diluição: BotuSêmen®; BotuSêmen® adicionado de 20 µg/mL de quercetina (SIGMA®); Solução aquosa com 10% leite em pó desnatado (Molico®) ou Solução aquosa com 10% leite em pó desnatado (Molico®) adicionado de 20 µg/mL de quercetina (SIGMA®). Por meio dos resultados obtidos no presente estudo, foi possível verificar que não há diferença estatística entre a motilidade espermática de sêmen de garanhão diluído em BotuSêmen® em relação ao diluído solução aquosa com 10% leite em pó desnatado (Molico®) pelo período de refrigeração avaliado (36h).(AU)

The biotechnique of equine semen cooling for the transfer of genetic material is widely distributed. With the need to maintain sperm viability, it is indispensable to use and thinners for this purpose. The aim of this study was to evaluate whether the addition of quercetin, which is an anti-oxidant flavonoid, in spermatic diluents alters the viability of refrigerated equine spermatozoa. Five ejaculates of a stallion were evaluated by analysis of motility and spermatic vigor in different cooling periods in four dilution media: BotuSêmen® ; BotuSêmen® added 20 µg/mL of quercetin (SIGMA® ); Aqueous solution with 10% skimmilk powder (Molico® ) or Aqueous solution with 10% skimmilk powder (Molico® ) added 20 µg/mL quercetin (SIGMA® ). Through the data obtained, it was possible to verify that there is no statistical difference between the sperm motility of diluted stallion semen in BotuSêmen® in relation to the diluted aqueous solution with 10% skimmed milk powder (Molico® ) for the evaluated refrigeration period (36h).(AU)

Effects of treatment with quercetin on the quality of cryopreserved bovine semen

Background: Semen cryopreservation is one of the most common biotechnologies in the reproduction of animals ofagricultural interest, especially bulls. However, cryopreservation can be harmful to sperm cells, with susceptibility tooxidative stress being one of the causes. The addition of antioxidants such as quercetin may inhibit and/or reduce suchdamage, reducing fertility. Quercetin can increasing sperm motility and interaction capacity between spermatozoa-oocyte,to increase cellular metabolism and reduced DNA fragmentation and oxidation following thawing. Therefore, the objective of this study was to evaluate the protective effect of quercetin on the metabolism of bovine semen following thawing.Materials, Methods & Results: Three Brahman bulls in reproduction age and previously considered fit for reproductionwere used. The semen samples were collected via the electroejaculation method, and the samples were homogenized toform pooled semen from three ejaculates, which was diluted in Tris-yolk egg-glicerol diluent medium. Quercetin was addedto diluent, to final concentrations of 0, 5, 10, 15 and 20 μg.mL-1 in each group. The samples were kept frozen in strawsof 500 μL, with concentration of 40,000,000 spermatozoid / mL for 15 days and were thawed in water at 36°C for 30 s.All the tests was performed in five replicates. The cell metabolism status was evaluated by quantification of superoxideradical production with a nitroblue tetrazolium test (NBT) and scanning spectrophotometry. By spermatic evaluation, thefollowing parameters were evaluated via the computerized system of sperm analysis (CASA): total motility (TM, %),progressive motility (PM, %), velocity curved line (VCL, mm/s), velocity straight line (VSL, mm/s), velocity averagepath (VAP, mm/s), distance curved line (DCL, mm), distance straight line (DSL, mm), distance average path (DAP, mm),amplitude of lateral head displacement (ALH, mm), beat cross frequency (BCF, Hz), wobble...

Effects of treatment with quercetin on the quality of cryopreserved bovine semen

Background: Semen cryopreservation is one of the most common biotechnologies in the reproduction of animals ofagricultural interest, especially bulls. However, cryopreservation can be harmful to sperm cells, with susceptibility tooxidative stress being one of the causes. The addition of antioxidants such as quercetin may inhibit and/or reduce suchdamage, reducing fertility. Quercetin can increasing sperm motility and interaction capacity between spermatozoa-oocyte,to increase cellular metabolism and reduced DNA fragmentation and oxidation following thawing. Therefore, the objective of this study was to evaluate the protective effect of quercetin on the metabolism of bovine semen following thawing.Materials, Methods & Results: Three Brahman bulls in reproduction age and previously considered fit for reproductionwere used. The semen samples were collected via the electroejaculation method, and the samples were homogenized toform pooled semen from three ejaculates, which was diluted in Tris-yolk egg-glicerol diluent medium. Quercetin was addedto diluent, to final concentrations of 0, 5, 10, 15 and 20 μg.mL-1 in each group. The samples were kept frozen in strawsof 500 μL, with concentration of 40,000,000 spermatozoid / mL for 15 days and were thawed in water at 36°C for 30 s.All the tests was performed in five replicates. The cell metabolism status was evaluated by quantification of superoxideradical production with a nitroblue tetrazolium test (NBT) and scanning spectrophotometry. By spermatic evaluation, thefollowing parameters were evaluated via the computerized system of sperm analysis (CASA): total motility (TM, %),progressive motility (PM, %), velocity curved line (VCL, mm/s), velocity straight line (VSL, mm/s), velocity averagepath (VAP, mm/s), distance curved line (DCL, mm), distance straight line (DSL, mm), distance average path (DAP, mm),amplitude of lateral head displacement (ALH, mm), beat cross frequency (BCF, Hz), wobble...(AU)

Alternativas fitoterápicas para o tratamento da leishmaniose

[...] Na procura de novos compostos com atividade leishmanicida destacam-se os alcalóides como a classe que tem maior número de compostos, dentre outros como os acetogeninas, flavonóides e componentes de óleos essenciais. Neste estudo foram utilizados alcalóides e acetogeninas extraídos da semente da Annona muricata (graviola); rutina e quercetina isolados das sementes Dimorphandra gardneriana (faveira), o eugenol, timol e seus derivados sintéticos, além da cumarina isolada do caule e cerne Platymiscium floribundum (sacambu). Foi realizado um screening in vitro com as formas promastigotas e amastigotas de L. i. chagasi, L. major, L. donovani e L. mexicana. Utilizou-se o método colorimétrico MTT ou cepas dependentes de Luciferase para a avaliação da efetividade das substâncias em promastigotas. E para a avaliação em amastigotas foi utilizado o método de leishmania in situ. Para os testes in vivo foram utilizados camundongos BALB/c infectados com promastigotas de L. i. chagasi na concentração de 107, via intraperitoneal. Os animais foram divididos em grupos de cinco, e tratados com 100 ?g/Kg dos derivados do eugenol e timol assim com as acetogeninas isoladas das sementes da graviola. Após 30 dias da infecção deu-se inicío ao tratamento em todos os grupos. Para o controle positivo foi utilizando um veículo de solubilização por via oral e para o grupo negativo foi utilizado o antimonial N-metil-glucamina administrado 60mg/kg/dia por via intramuscular durante 30 dias. Os grupos de animais tratados e não tratados foram eutanasiados após 30 dias de tratamento, e retirados, de maneira asséptica, o baço e o fígado, para a determinação da carga parasitária e para os estudos posteriores da quantificação da parasitemia por qPCR.

[...] Alkaloids stand out in search of new compounds with leishmanicidal activity , as a class with the greatest number of compounds, among others like acetogenins, flavonoids and components of Essential Oils. In this study, alkaloids and acetogenins extracted from the seed of Annona muricata (soursop) , rutin and quercetin isolated from the seeds of Dimorphandra gardneriana (faveira) , eugenol, thymol and their synthetic derivates, in addition of coumarin, isolated from the trunk heartwood Platymiscium floribundum (sacambu) were used. In vitro Screening was conducted with promastigotes an d amastigotes forms of L. i. chagasi , L. major , L. donovani and L. mexicana, using either the MTT colorimetric method or Luciferase dependent strains to the evaluation of these studied substances effectiveness against promastigotes, and the in situ leishma nia to the assessment against amastigotes. To the in vivo tests BALB/c infected with promastigotes of L. i. chagasi were used at the concentration of 10 7 intrapetitoneally. The animal were divided into groups of five, and treated with 100 μg/Kg of thymol a nd eugenol derivatives, and the acetogenins isolated from graviola. Thrity days post infections began the treatment in all groups. As positive control an oral solubilization vehicle was used and to the negative group N - methyl - glucamine antimony 60mg/kg/d ay administered intramuscularly for 30 days. The treated and not treated groups were euthanized after 30 days of treatment, and had spleen and liver aseptically removed, to the determination of parasite load and further studies of the measurement of parasitemia by qPCR.

Alternativas fitoterápicas para o tratamento da leishmaniose

[...] Na procura de novos compostos com atividade leishmanicida destacam-se os alcalóides como a classe que tem maior número de compostos, dentre outros como os acetogeninas, flavonóides e componentes de óleos essenciais. Neste estudo foram utilizados alcalóides e acetogeninas extraídos da semente da Annona muricata (graviola); rutina e quercetina isolados das sementes Dimorphandra gardneriana (faveira), o eugenol, timol e seus derivados sintéticos, além da cumarina isolada do caule e cerne Platymiscium floribundum (sacambu). Foi realizado um screening in vitro com as formas promastigotas e amastigotas de L. i. chagasi, L. major, L. donovani e L. mexicana. Utilizou-se o método colorimétrico MTT ou cepas dependentes de Luciferase para a avaliação da efetividade das substâncias em promastigotas. E para a avaliação em amastigotas foi utilizado o método de leishmania in situ. Para os testes in vivo foram utilizados camundongos BALB/c infectados com promastigotas de L. i. chagasi na concentração de 107, via intraperitoneal. Os animais foram divididos em grupos de cinco, e tratados com 100 ?g/Kg dos derivados do eugenol e timol assim com as acetogeninas isoladas das sementes da graviola. Após 30 dias da infecção deu-se inicío ao tratamento em todos os grupos. Para o controle positivo foi utilizando um veículo de solubilização por via oral e para o grupo negativo foi utilizado o antimonial N-metil-glucamina administrado 60mg/kg/dia por via intramuscular durante 30 dias. Os grupos de animais tratados e não tratados foram eutanasiados após 30 dias de tratamento, e retirados, de maneira asséptica, o baço e o fígado, para a determinação da carga parasitária e para os estudos posteriores da quantificação da parasitemia por qPCR.(AU)

[...] Alkaloids stand out in search of new compounds with leishmanicidal activity , as a class with the greatest number of compounds, among others like acetogenins, flavonoids and components of Essential Oils. In this study, alkaloids and acetogenins extracted from the seed of Annona muricata (soursop) , rutin and quercetin isolated from the seeds of Dimorphandra gardneriana (faveira) , eugenol, thymol and their synthetic derivates, in addition of coumarin, isolated from the trunk heartwood Platymiscium floribundum (sacambu) were used. In vitro Screening was conducted with promastigotes an d amastigotes forms of L. i. chagasi , L. major , L. donovani and L. mexicana, using either the MTT colorimetric method or Luciferase dependent strains to the evaluation of these studied substances effectiveness against promastigotes, and the in situ leishma nia to the assessment against amastigotes. To the in vivo tests BALB/c infected with promastigotes of L. i. chagasi were used at the concentration of 10 7 intrapetitoneally. The animal were divided into groups of five, and treated with 100 μg/Kg of thymol a nd eugenol derivatives, and the acetogenins isolated from graviola. Thrity days post infections began the treatment in all groups. As positive control an oral solubilization vehicle was used and to the negative group N - methyl - glucamine antimony 60mg/kg/d ay administered intramuscularly for 30 days. The treated and not treated groups were euthanized after 30 days of treatment, and had spleen and liver aseptically removed, to the determination of parasite load and further studies of the measurement of parasitemia by qPCR.(AU)