ATIVIDADE FITOESTROGÊNICA DE Morus nigra L., MORACEAE, EM RATAS OVARIECTOMIZADAS

Detalhes bibliográficos
Autor(a) principal: SILVA, Selma do Nascimento
Data de Publicação: 2012
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFMA
Texto Completo: http://tedebc.ufma.br:8080/jspui/handle/tede/68
Resumo: The hypoestrogenism in climacteric is associated with vasomotor symptoms, cardiovascular disease, osteoporosis and urogenital changes. At this stage of life of women, hormone replacement HRT can alleviate some consequences of decreased estrogen caused by ovarian failure. However, estrogen therapy may cause adverse effects such as breast tenderness, uterine bleeding and increase the relative risk for cancers of the breast and endometrium. Morus nigra L. (mulberry) is a plant species most used in Brazil for the treatment of menopausal symptoms. Thus, this study aims to assess the likely effects phytoestrogenics the hydroalcoholic extract (HE) from the leaves of M. nigra in ovariectomized female rats. Therefore, the dried leaves were pulverized and soaked in 70% ethanol in the proportion 1:3 (v/v) to obtain the HE (yield=21.90%). The HE was subjected to evaluation of the antioxidant activity by capturing free radical 2,2-diphenyl-1-picryl-hidrazil, analyzed by liquid chromatography coupled to mass spectrum (LC-MS/MS) for identification of compound and then partitioned with hexane, chloroform, ethyl acetate and methanol/water. The safety of extract was determined by the test of acute toxicity in mice at doses of 0.1 to 10.0 g/kg orally (p.o.). To evaluate the estrogenic activity of the extract from M. nigra leaves, the rats were divided into two control groups: sham-operated (SHAM) and ovariectomized (OVX), which received 0.1 mL/100 g saline, and two test groups: ovariectomized and treated with a solution estroprogestative (OVX-EP-50g/kg) and ovariectomized and treated with HE M. nigra 500mg/kg (OVX-HE500), n = 8-10, daily, p.o., for 14 weeks. Throughout the treatment period were analyzed the frequency stage of the estrous cycle, food intake and body weight. At the end of treatment were evaluated biochemical parameters and hormone, histomorphometry of the uterus, vagina and breast. Furthermore, the influence of M. nigra on the proliferation of breast tumor cell line MCF-7 was determined by MTT method. HE showed high antioxidant activity when compared to standard quercetin. The analysis by LC-MS/MS EH compared with literature data allowed the identification of flavonoids (kaempferol and quercetin) and quinic acid derivatives (caffeoylquinic acid and isomers dicaffeoylquinic acid). In the analysis of the estrous cycle, the group OVX-HE500 showed an increase in proestrous and estrous phases at 15.25% and 26.6%, respectively, when compared to OVX group. Ovariectomy caused an increase in body mass, which was prevented by treatment with HE and EP solution. The weight of abdominal adipose tissue was also significantly lower in groups OVX-HE and OVX-EP compared to the OVX group. Ovariectomy also induced atrophy of uterine tissue (OVX group) compared to SHAM group, indicating the efficiency of the surgical procedure, and the administration of EP significantly increased uterine weight compared with OVX group. Average uterine weight of the OVX-HE group was also higher than the OVX group, but smaller than the OVX-EP group. In the histological analysis, it was observed that the characteristics of the squamous epithelium of the vagina of OVX-EP group (57.79 ± 1.49m), relative to thickness, were similar to that of SHAM group (50 66 ± 1.60m). After 14 weeks of administration of HE was a partial reversal of vaginal atrophy (37.34 ± 1.77m), when compared to the OVX group (12.92 ± 0.53m), showing maturation of this tissue with the treatment, however, the HE did not alter breast tissue, unlike the stimulus EP-induced. Regarding biochemistry was observed that the treatments (HE and EP) reduced concentrations of triglycerides in 27.5% and 23.8% respectively, when compared to OVX. In in vitro tests, the data indicate that the HE M. nigra acts as a weak phytoestrogen and protects against cell proliferation of human breast carcinoma (MCF-7). In acute toxicity study, the treatment of mice with HE did not produce behavioral changes or deaths. Together, the data demonstrate that the HE M. nigra L. has beneficial effects in models of induced menopause in rats, decreased uterine and vaginal atrophy, without changing the mammary structure, improving triglyceride levels and shows up secure and potent oxidant activity. These effects may be related to their flavonoid constituents, and thus the plant species may be useful in controlling symptoms of menopause as an alternative to Hormone Replacement Therapy.
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spelling BORGES, Antônio Carlos RomãoCPF:54858723510http://lattes.cnpq.br/4315209704773266BORGES, Marilene Oliveira da RochaCPF:22729860215http://lattes.cnpq.br/3283574594616838CPF:14758725415http://lattes.cnpq.br/0400234542205258SILVA, Selma do Nascimento2016-08-16T18:18:42Z2015-03-102012-10-02http://tedebc.ufma.br:8080/jspui/handle/tede/68The hypoestrogenism in climacteric is associated with vasomotor symptoms, cardiovascular disease, osteoporosis and urogenital changes. At this stage of life of women, hormone replacement HRT can alleviate some consequences of decreased estrogen caused by ovarian failure. However, estrogen therapy may cause adverse effects such as breast tenderness, uterine bleeding and increase the relative risk for cancers of the breast and endometrium. Morus nigra L. (mulberry) is a plant species most used in Brazil for the treatment of menopausal symptoms. Thus, this study aims to assess the likely effects phytoestrogenics the hydroalcoholic extract (HE) from the leaves of M. nigra in ovariectomized female rats. Therefore, the dried leaves were pulverized and soaked in 70% ethanol in the proportion 1:3 (v/v) to obtain the HE (yield=21.90%). The HE was subjected to evaluation of the antioxidant activity by capturing free radical 2,2-diphenyl-1-picryl-hidrazil, analyzed by liquid chromatography coupled to mass spectrum (LC-MS/MS) for identification of compound and then partitioned with hexane, chloroform, ethyl acetate and methanol/water. The safety of extract was determined by the test of acute toxicity in mice at doses of 0.1 to 10.0 g/kg orally (p.o.). To evaluate the estrogenic activity of the extract from M. nigra leaves, the rats were divided into two control groups: sham-operated (SHAM) and ovariectomized (OVX), which received 0.1 mL/100 g saline, and two test groups: ovariectomized and treated with a solution estroprogestative (OVX-EP-50g/kg) and ovariectomized and treated with HE M. nigra 500mg/kg (OVX-HE500), n = 8-10, daily, p.o., for 14 weeks. Throughout the treatment period were analyzed the frequency stage of the estrous cycle, food intake and body weight. At the end of treatment were evaluated biochemical parameters and hormone, histomorphometry of the uterus, vagina and breast. Furthermore, the influence of M. nigra on the proliferation of breast tumor cell line MCF-7 was determined by MTT method. HE showed high antioxidant activity when compared to standard quercetin. The analysis by LC-MS/MS EH compared with literature data allowed the identification of flavonoids (kaempferol and quercetin) and quinic acid derivatives (caffeoylquinic acid and isomers dicaffeoylquinic acid). In the analysis of the estrous cycle, the group OVX-HE500 showed an increase in proestrous and estrous phases at 15.25% and 26.6%, respectively, when compared to OVX group. Ovariectomy caused an increase in body mass, which was prevented by treatment with HE and EP solution. The weight of abdominal adipose tissue was also significantly lower in groups OVX-HE and OVX-EP compared to the OVX group. Ovariectomy also induced atrophy of uterine tissue (OVX group) compared to SHAM group, indicating the efficiency of the surgical procedure, and the administration of EP significantly increased uterine weight compared with OVX group. Average uterine weight of the OVX-HE group was also higher than the OVX group, but smaller than the OVX-EP group. In the histological analysis, it was observed that the characteristics of the squamous epithelium of the vagina of OVX-EP group (57.79 ± 1.49m), relative to thickness, were similar to that of SHAM group (50 66 ± 1.60m). After 14 weeks of administration of HE was a partial reversal of vaginal atrophy (37.34 ± 1.77m), when compared to the OVX group (12.92 ± 0.53m), showing maturation of this tissue with the treatment, however, the HE did not alter breast tissue, unlike the stimulus EP-induced. Regarding biochemistry was observed that the treatments (HE and EP) reduced concentrations of triglycerides in 27.5% and 23.8% respectively, when compared to OVX. In in vitro tests, the data indicate that the HE M. nigra acts as a weak phytoestrogen and protects against cell proliferation of human breast carcinoma (MCF-7). In acute toxicity study, the treatment of mice with HE did not produce behavioral changes or deaths. Together, the data demonstrate that the HE M. nigra L. has beneficial effects in models of induced menopause in rats, decreased uterine and vaginal atrophy, without changing the mammary structure, improving triglyceride levels and shows up secure and potent oxidant activity. These effects may be related to their flavonoid constituents, and thus the plant species may be useful in controlling symptoms of menopause as an alternative to Hormone Replacement Therapy.O hipoestrogenismo no climatério associa-se com sintomas vasomotores, doenças cardiovasculares, osteoporose e alterações urogenitais. Nesta fase da vida da mulher, a reposição hormonal pode amenizar algumas consequências da diminuição estrogênica ocasionada pela falência ovariana. Porém, a terapia estrogênica pode ocasionar efeitos adversos como mastalgia, sangramentos uterinos, além de aumentar o risco relativo para neoplasias de mama e endométrio. Morus nigra L. (amora) é uma das espécies vegetais mais utilizadas no Brasil para o tratamento dos sintomas do climatério. Assim, o presente estudo objetiva avaliar os prováveis efeitos fitoestrogênicos do extrato hidroalcoólico (EH) das folhas de M. nigra em ratas Wistar ovariectomizadas. Para tanto, as folhas secas foram pulverizadas e maceradas em etanol a 70% na proporção 1:3 (v/v), para obtenção do EH (rendimento=21,90%). O EH foi submetido à avaliação da atividade antioxidante pela captura do radical livre 2,2-difenil-1-picril-hidrazila, analisado por cromatografia líquida acoplada ao espectro de massa (LC-MS/MS) para identificação de composto e, em seguida, particionado com hexano, clorofórmio, acetato de etila e metanol/água. A segurança do extrato foi determinada pelo teste de toxidade aguda em camundongos, nas doses de 0,1 10,0g/kg, por via oral (v.o.). Para avaliar a atividade estrogênica do extrato das folhas de M. nigra, as ratas foram divididas em dois grupos controle: falso-operados (SHAM) e ratas ovariectomizadas (OVX), que receberam 0,1mL/100g de solução salina; e dois grupos teste: ovariectomizadas e tratadas com solução estroprogestativa (OVX-EP-50g/Kg) e ovariectomizadas e tratadas com EH de M. nigra 500mg/kg (OVX-EH500), n=8-10, diariamente, por v.o., durante 14 semanas. Durante todo o período de tratamento foram analisadas a frequência das fases do ciclo estral, a ingestão de alimentos e o peso corporal. Ao final do tratamento foram avaliados os parâmetros bioquímicos e hormonais, histomorfometria do útero, vagina e mama. Além disso, a influência de M. nigra sobre a proliferação de células tumorais de mama da linhagem MCF-7 foi determinada pelo método MTT. O EH apresentou alta atividade antioxidante quando comparada ao padrão quercetina. Na análise do EH por LC-MS/MS em comparação com dados da literatura permitiu a identificação de flavonoides (caempferol e quercetina) e derivados do ácido quínico (ácido cafeoilquínico e isômeros de ácido dicafeoilquínico). Na análise do ciclo estral, o grupo OVX-EH500 apresentou um aumento nas fases estro e proestro em 15,25% e 26,6%, respectivamente, quando comparado ao grupo OVX. A ovariectomia promoveu um aumento no peso corporal, que foi inibido pelo tratamento com o EH e solução EP. O peso do tecido adiposo abdominal também foi significativamente menor nos grupos OVX-EP e OVX-EH, quando comparados ao grupo OVX. A ovariectomia também induziu atrofia do tecido uterino (Grupo OVX) em comparação ao grupo SHAM, indicando a eficiência do procedimento cirúrgico; e a administração de EP aumentou significativamente o peso do útero em comparação com grupo OVX. A média do peso uterino do grupo OVX-EH também foi maior do que o grupo OVX, porém menor que o grupo OVX-EP. Quanto à análise histológica, observou-se que as características do epitélio escamoso da vagina do grupo OVX-EP (57,79 ± 1,49m), em relação à espessura, se assemelharam à das ratas do grupo SHAM (50,66 ± 1,60m). Após 14 semanas de administração de EH houve uma reversão parcial da atrofia vaginal (37,34 ± 1,77m), quando comparado ao grupo OVX (12,92 ± 0,53 m), mostrando maturação deste tecido com o tratamento; entretanto, o EH não alterou o tecido mamário, diferente do estímulo induzido pelo EP. Em relação à bioquímica foi observado que os tratamentos (EH e EP) reduziram as concentrações de triglicérides em 27,5% e 23,8% respectivamente, quando comparado ao grupo OVX. Nos testes in vitro, os dados indicam que o EH de M. nigra atua como um fraco fitoestrógeno e protege contra a proliferação de células de carcinoma de mama humano (MCF-7). No estudo toxicológico agudo, o tratamento de camundongos com o EH não produziu alterações comportamentais nem mortes. Em conjunto, os dados demonstram que o EH de M.nigra apresenta efeitos benéficos em modelos de hipoestrogenismo induzida em ratas, diminuindo a atrofia uterina e vaginal, sem alterar a estrutura mamária, melhorando os níveis de triglicérides, tendo potencial antioxidante, além de mostrar-se seguro. Esses efeitos podem estar relacionados com seus constituintes flavonoídicos, e dessa forma, a espécie vegetal pode ser útil no controle de sintomas da menopausa como uma alternativa para Terapia de Reposição Hormonal.Made available in DSpace on 2016-08-16T18:18:42Z (GMT). No. of bitstreams: 1 TESE SELMA DO NASCIMENTO SILVA.pdf: 3613537 bytes, checksum: e7528662a8a3d2ec2dc6bf7166cc0e52 (MD5) Previous issue date: 2012-10-02application/pdfporUniversidade Federal do MaranhãoPROGRAMA DE PÓS-GRADUAÇÃO EM BIOTECNOLOGIA - RENORBIO/CCBSUFMABRFertilizaçãoMorus nigra L.amora-pretaterapia hormonalflavonoidesMorus nigra L.hormone therapyovariectomized ratsflavonoidsLC-MS/MSCIENCIAS BIOLOGICAS::FARMACOLOGIAATIVIDADE FITOESTROGÊNICA DE Morus nigra L., MORACEAE, EM RATAS OVARIECTOMIZADASACTIVITY OF FITOESTROGÊNICA, MORUS NIGRA L. MORACEAE, IN RATS VARIECTOMIZEDinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações da UFMAinstname:Universidade Federal do Maranhão (UFMA)instacron:UFMASILVA, Selma do Nascimento. ATIVIDADE FITOESTROGÊNICA DE Morus nigra L., MORACEAE, EM RATAS OVARIECTOMIZADAS. 2012. 125 f. Tese( Programa de Pós-graduação em Biotecnologia - RENORBIO/CCBS) - Universidade Federal do Maranhão, São Luís, 2012.ORIGINALTESE SELMA DO NASCIMENTO SILVA.pdfapplication/pdf3613537http://tedebc.ufma.br:8080/bitstream/tede/68/1/TESE+SELMA+DO+NASCIMENTO+SILVA.pdfe7528662a8a3d2ec2dc6bf7166cc0e52MD51tede/682022-12-21 16:04:48.017oai:tede2:tede/68Biblioteca Digital de Teses e Dissertaçõeshttps://tedebc.ufma.br/jspui/PUBhttp://tedebc.ufma.br:8080/oai/requestrepositorio@ufma.br||repositorio@ufma.bropendoar:21312022-12-21T19:04:48Biblioteca Digital de Teses e Dissertações da UFMA - Universidade Federal do Maranhão (UFMA)false
dc.title.por.fl_str_mv ATIVIDADE FITOESTROGÊNICA DE Morus nigra L., MORACEAE, EM RATAS OVARIECTOMIZADAS
dc.title.alternative.eng.fl_str_mv ACTIVITY OF FITOESTROGÊNICA, MORUS NIGRA L. MORACEAE, IN RATS VARIECTOMIZED
title ATIVIDADE FITOESTROGÊNICA DE Morus nigra L., MORACEAE, EM RATAS OVARIECTOMIZADAS
spellingShingle ATIVIDADE FITOESTROGÊNICA DE Morus nigra L., MORACEAE, EM RATAS OVARIECTOMIZADAS
SILVA, Selma do Nascimento
Morus nigra L.
amora-preta
terapia hormonal
flavonoides
Morus nigra L.
hormone therapy
ovariectomized rats
flavonoids
LC-MS/MS
CIENCIAS BIOLOGICAS::FARMACOLOGIA
title_short ATIVIDADE FITOESTROGÊNICA DE Morus nigra L., MORACEAE, EM RATAS OVARIECTOMIZADAS
title_full ATIVIDADE FITOESTROGÊNICA DE Morus nigra L., MORACEAE, EM RATAS OVARIECTOMIZADAS
title_fullStr ATIVIDADE FITOESTROGÊNICA DE Morus nigra L., MORACEAE, EM RATAS OVARIECTOMIZADAS
title_full_unstemmed ATIVIDADE FITOESTROGÊNICA DE Morus nigra L., MORACEAE, EM RATAS OVARIECTOMIZADAS
title_sort ATIVIDADE FITOESTROGÊNICA DE Morus nigra L., MORACEAE, EM RATAS OVARIECTOMIZADAS
author SILVA, Selma do Nascimento
author_facet SILVA, Selma do Nascimento
author_role author
dc.contributor.advisor1.fl_str_mv BORGES, Antônio Carlos Romão
dc.contributor.advisor1ID.fl_str_mv CPF:54858723510
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/4315209704773266
dc.contributor.advisor-co1.fl_str_mv BORGES, Marilene Oliveira da Rocha
dc.contributor.advisor-co1ID.fl_str_mv CPF:22729860215
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/3283574594616838
dc.contributor.authorID.fl_str_mv CPF:14758725415
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/0400234542205258
dc.contributor.author.fl_str_mv SILVA, Selma do Nascimento
contributor_str_mv BORGES, Antônio Carlos Romão
BORGES, Marilene Oliveira da Rocha
dc.subject.por.fl_str_mv Morus nigra L.
amora-preta
terapia hormonal
flavonoides
topic Morus nigra L.
amora-preta
terapia hormonal
flavonoides
Morus nigra L.
hormone therapy
ovariectomized rats
flavonoids
LC-MS/MS
CIENCIAS BIOLOGICAS::FARMACOLOGIA
dc.subject.eng.fl_str_mv Morus nigra L.
hormone therapy
ovariectomized rats
flavonoids
LC-MS/MS
dc.subject.cnpq.fl_str_mv CIENCIAS BIOLOGICAS::FARMACOLOGIA
description The hypoestrogenism in climacteric is associated with vasomotor symptoms, cardiovascular disease, osteoporosis and urogenital changes. At this stage of life of women, hormone replacement HRT can alleviate some consequences of decreased estrogen caused by ovarian failure. However, estrogen therapy may cause adverse effects such as breast tenderness, uterine bleeding and increase the relative risk for cancers of the breast and endometrium. Morus nigra L. (mulberry) is a plant species most used in Brazil for the treatment of menopausal symptoms. Thus, this study aims to assess the likely effects phytoestrogenics the hydroalcoholic extract (HE) from the leaves of M. nigra in ovariectomized female rats. Therefore, the dried leaves were pulverized and soaked in 70% ethanol in the proportion 1:3 (v/v) to obtain the HE (yield=21.90%). The HE was subjected to evaluation of the antioxidant activity by capturing free radical 2,2-diphenyl-1-picryl-hidrazil, analyzed by liquid chromatography coupled to mass spectrum (LC-MS/MS) for identification of compound and then partitioned with hexane, chloroform, ethyl acetate and methanol/water. The safety of extract was determined by the test of acute toxicity in mice at doses of 0.1 to 10.0 g/kg orally (p.o.). To evaluate the estrogenic activity of the extract from M. nigra leaves, the rats were divided into two control groups: sham-operated (SHAM) and ovariectomized (OVX), which received 0.1 mL/100 g saline, and two test groups: ovariectomized and treated with a solution estroprogestative (OVX-EP-50g/kg) and ovariectomized and treated with HE M. nigra 500mg/kg (OVX-HE500), n = 8-10, daily, p.o., for 14 weeks. Throughout the treatment period were analyzed the frequency stage of the estrous cycle, food intake and body weight. At the end of treatment were evaluated biochemical parameters and hormone, histomorphometry of the uterus, vagina and breast. Furthermore, the influence of M. nigra on the proliferation of breast tumor cell line MCF-7 was determined by MTT method. HE showed high antioxidant activity when compared to standard quercetin. The analysis by LC-MS/MS EH compared with literature data allowed the identification of flavonoids (kaempferol and quercetin) and quinic acid derivatives (caffeoylquinic acid and isomers dicaffeoylquinic acid). In the analysis of the estrous cycle, the group OVX-HE500 showed an increase in proestrous and estrous phases at 15.25% and 26.6%, respectively, when compared to OVX group. Ovariectomy caused an increase in body mass, which was prevented by treatment with HE and EP solution. The weight of abdominal adipose tissue was also significantly lower in groups OVX-HE and OVX-EP compared to the OVX group. Ovariectomy also induced atrophy of uterine tissue (OVX group) compared to SHAM group, indicating the efficiency of the surgical procedure, and the administration of EP significantly increased uterine weight compared with OVX group. Average uterine weight of the OVX-HE group was also higher than the OVX group, but smaller than the OVX-EP group. In the histological analysis, it was observed that the characteristics of the squamous epithelium of the vagina of OVX-EP group (57.79 ± 1.49m), relative to thickness, were similar to that of SHAM group (50 66 ± 1.60m). After 14 weeks of administration of HE was a partial reversal of vaginal atrophy (37.34 ± 1.77m), when compared to the OVX group (12.92 ± 0.53m), showing maturation of this tissue with the treatment, however, the HE did not alter breast tissue, unlike the stimulus EP-induced. Regarding biochemistry was observed that the treatments (HE and EP) reduced concentrations of triglycerides in 27.5% and 23.8% respectively, when compared to OVX. In in vitro tests, the data indicate that the HE M. nigra acts as a weak phytoestrogen and protects against cell proliferation of human breast carcinoma (MCF-7). In acute toxicity study, the treatment of mice with HE did not produce behavioral changes or deaths. Together, the data demonstrate that the HE M. nigra L. has beneficial effects in models of induced menopause in rats, decreased uterine and vaginal atrophy, without changing the mammary structure, improving triglyceride levels and shows up secure and potent oxidant activity. These effects may be related to their flavonoid constituents, and thus the plant species may be useful in controlling symptoms of menopause as an alternative to Hormone Replacement Therapy.
publishDate 2012
dc.date.issued.fl_str_mv 2012-10-02
dc.date.available.fl_str_mv 2015-03-10
dc.date.accessioned.fl_str_mv 2016-08-16T18:18:42Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
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dc.identifier.uri.fl_str_mv http://tedebc.ufma.br:8080/jspui/handle/tede/68
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dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
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dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal do Maranhão
dc.publisher.program.fl_str_mv PROGRAMA DE PÓS-GRADUAÇÃO EM BIOTECNOLOGIA - RENORBIO/CCBS
dc.publisher.initials.fl_str_mv UFMA
dc.publisher.country.fl_str_mv BR
dc.publisher.department.fl_str_mv Fertilização
publisher.none.fl_str_mv Universidade Federal do Maranhão
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