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Chemical and biological characterization of a steroidogenic protein from Dioscorea opposita ThunbWong, Kam-lok, 黃金樂 January 2011 (has links)
Menopause is the period during which the level of estrogen secreted by the ovaries gradually declines and patients of menopausal syndrome may experience osteoporosis, cognitive decline, hot flush, mood disorder, night sweat, depression, nervous tension and insomnia. The report published by the World Health Organization in 1990 stated that the total population of postmenopausal women in the world was 476 million. By 2030, the predicted population will reach 1200 million. The current way to relieve menopausal syndrome is hormone replacement therapy (HRT) which restores the endogenous estrogen level by administration of supplementary exogenous estrogen or estrogen plus progestin. However, various studies showed that HRT might increase the incidence of breast cancer and ovarian cancer. There is a potential risk of using HRT for menopausal syndrome.
The novel protein DO isolated in this study could be a potential alternative of HRT for the treatment of menopausal syndrome. DO isolated from Chinese yam Dioscorea opposite Thunb. had adistinctive N-terminal sequence Gly-Ile-Gly-Lys-Ile-Thr-Thr-
Tyr-Trp-Gly-Gln-Tyr-Ser-Asp-Glu-Pro-Ser-Leu-Thr-Glu-Ala, indicating that DO might be a novel protein.DO was an acid stable and thermal stable protein. As determined by electro-chemiluminescence immunoassay (ECLIA) detection, DO could stimulate estradiol biosynthesis in rat granulosa cells in vitro as well as inducing both estradiol and progesterone secretion in female aging Sprague Darley (SD)-rat model in vivo. From the results of real-time PCR and Western blot, the possible mechanism might be through up-regulating the expression of ovarian follicle stimulating hormone receptor(FSHR)and aromatase. DO also exerted a beneficial effect on the cognitive function through elevating the translational level of brain derived neurotrophic factor (BDNF)and TrkB gp 145 receptor in the prefrontal cortex of female aging SD-rat model.
Though further pharmacological studies are required, the results in this study suggested that DO could be a safer potential alternative for HRT in the treatment of menopausal syndrome with beneficial effects on hormone levels and cognitive function. / published_or_final_version / Chinese Medicine / Master / Master of Philosophy
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Erxian decoction for menopause: systematic review and mechanistic study in estradiol bio-synthesis in vitroChen, Haiyong., 陳海勇. January 2008 (has links)
published_or_final_version / Chinese Medicine / Master / Master of Philosophy
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Elucidation of the action mechanism of erxian decoction, a Chinese medicinal formula for menopause: frompharmacological approach to analytical approachCheung, Ho-pan., 張浩斌. January 2013 (has links)
As the aging in reproductive system proceeds, females will eventually enter the period of menopause, during which a series of physiological changes occurs. The decline of estrogen level during menopausal transition is thought to associate with various menopausal symptoms. Although hormone replacement therapy can be adopted to deal with the estrogen-deficient state, side effects such as cancer risk cannot be overlooked.
Alternatively, Erxian Decoction (EXD), a Chinese medicine formula for treating menopausal symptoms has been used clinically for more than 60 years without adverse effects reported. Some pharmacological properties of EXD have been reported in previous research, which are thought to be contributed by its multiple bioactive components. Thus in the present study, the pharmacological properties of EXD have been further evaluated. The drug compatibility of Traditional Chinese Medicine (TCM) formula, EXD, was also demonstrated. At last, a novel approach for identification of bioactive components from Chinese medicine formula was introduced using EXD as study model.
To evaluate the multiple pharmacological properties of EXD, proteins involved in steroidogenesis in ovaries of aged female rats were measured by immunoblotting analysis. On top of that, serum lipid profiles and the related proteins were determined by colorimetric assay and immunoblotting analysis respectively. Also, anti-osteoporotic properties and drug compatibility of EXD were evaluated by in vitro methods such as proliferation assay, osteoclast differentiation assay, ELISA assay or immunoblotting analysis. Lastly, a novel approach for identification of bioactive components in relation to the subsequent bioactivity from traditional Chinese medicinal formula was introduced using HPLC profiles.
From the results, it was demonstrated that EXD can modulate steroidogenesis in aged female rat model at least through up-regulation of ovarian aromatase, protein kinase B and estrogen receptor beta at protein level. Besides, EXD also exerts antihyperlipidemic effects in aged female rats as reflected from the decreased serum total cholesterol and LDL-cholesterol levels via regulation of HMG CoA reductase and LDL-receptor, the key proteins for cholesterol synthesis and LDL-cholesterol clearance. In vitro study has also demonstrated the anti-osteoporotic properties of EXD through stimulation of osteoblast proliferation and inhibition of proliferation and differentiation of osteoclast precursor cells. The later was proved to be mediated by down-regulation of NFATc1 proteins, a key protein for osteoclastogenesis. The roles of the drugs categories according to the drug compatibility of traditional Chinese medicine contributing to the optimal anti-osteoporotic properties of EXD were also elucidated.
Since the diverse pharmacological properties of a Chinese medicinal formula are often the results of the effects of complex bioactive constituents in the extract, yet identification of the bioactive components has been a tedious task. Thus in the last part of the study, a novel approach for identification of bioactive component from Chinese medicinal formula has been developed. By comparing the HPLC profiles of EXD extracted by different decoction method in relation to their pharmacological properties, six bioactive chemicals were successfully identified which may contribute to the stimulatory effect of EXD on ovarian aromatase and hepatic catalase expression. / published_or_final_version / Chinese Medicine / Master / Master of Philosophy
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Determinação do 17 β-estradiol em formas farmacêuticas utilizadas na terapia de reposição hormonal / Determination of 17 β-estradiol in dosage forms used in hormone replacement therapyYakabe, Clarice 30 September 2004 (has links)
Atualmente, na terapia de reposição hormonal (TRH), o estrogênio é o tratamento de escolha para os sintomas da menopausa. Para o tratamento adequado, um dos estrogênios amplamente usado em TRH é o 17β-estradiol, o qual está disponível em diversas formas farmacêuticas. Um método espectrofotométrico na região do visível foi desenvolvido para quantificação do 17β-estradiol em gel, comprimido e adesivo transdérmico. O método baseou - se na reação de diazotação da sulfanilamida seguida de acoplamento ao 17β-estradiol, produzindo um azo composto de cor marrom - avermelhada. O comportamento da reação foi verificado quanto à concentração dos reagentes, espectro de absorção, estabilidade do produto colorido. A linearidade foi observada no intervalo de concentração entre 10,0 a 28,0 µg/mL, com um coeficiente de correlação de 0,9996. O limite de detecção foi de 0,76 µg/mL e o limite de quantificação, de 2,30 µg/mL. O método padronizado foi aplicado na amostra comercial e simulado, e os resultados do desvio padrão relativo foram abaixo de 2%, e a médias da porcentagem de recuperação do padrão variaram de 98 a 101%. O método colori métrico demonstrou ser preciso e exato, podendo ser usado para determinação quantitativa do 17β-estradiol nas formas farmacêuticas estudadas. No método espectrofotométrico na região do UV, o 17β-estradiol foi extraído com etanol absoluto. Determinados os parâmetros analíticos, o método mostrou-se muito simples, rápido, preciso, exato e mais econômico que o método colorimétrico, porém sendo aplicável somente aos comprimidos. / Nowadays, estrogen - based hormonal replacement therapy (HRT) is the treatment of choice for menopause symptoms. For adequate treatment, one of the most widely used estrogens in HRT is 17β-estradiol, which is available in several pharmaceutical dosage forms. A visible spectrophotometric method was developed for quantification of 17β-estradiol in gel, tablets and patches. The method was based on the diazotization reaction of sulfanilamide and azo coupling with 17β-estradiol, forming the brown - reddish azo compound. The reaction behavior was observed for reagent concentration, absorption spectra and color stability. The linearity of the method was observed in a concentration range from 10,0 to 28,0 µg/mL, with correlation coefficient of 0,9996. The detection limit was 0,76 µg/mL and the quantitation limit was 2,30 µg/mL. The standardized method was applied to dosage formulations and simulated samples; the relative standard deviations (RSD) were below of 2% and the average percentage recoveries of standards varied from 98 to 101%. The colorimetric method showed precise, accurate and inexpensive, and can be used for quantitative determination of 17β-estradiol in pharmaceutical formulations studied. On the UV spectrophometric method, the 17β-estradiol was extracted with ethanol absolute. After determining analytical parameters, this method showed very simple, fast, precise, accurate and more economic than colorimetric method but only be applied for tablet formulations.
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Determinação do 17 β-estradiol em formas farmacêuticas utilizadas na terapia de reposição hormonal / Determination of 17 β-estradiol in dosage forms used in hormone replacement therapyClarice Yakabe 30 September 2004 (has links)
Atualmente, na terapia de reposição hormonal (TRH), o estrogênio é o tratamento de escolha para os sintomas da menopausa. Para o tratamento adequado, um dos estrogênios amplamente usado em TRH é o 17β-estradiol, o qual está disponível em diversas formas farmacêuticas. Um método espectrofotométrico na região do visível foi desenvolvido para quantificação do 17β-estradiol em gel, comprimido e adesivo transdérmico. O método baseou - se na reação de diazotação da sulfanilamida seguida de acoplamento ao 17β-estradiol, produzindo um azo composto de cor marrom - avermelhada. O comportamento da reação foi verificado quanto à concentração dos reagentes, espectro de absorção, estabilidade do produto colorido. A linearidade foi observada no intervalo de concentração entre 10,0 a 28,0 µg/mL, com um coeficiente de correlação de 0,9996. O limite de detecção foi de 0,76 µg/mL e o limite de quantificação, de 2,30 µg/mL. O método padronizado foi aplicado na amostra comercial e simulado, e os resultados do desvio padrão relativo foram abaixo de 2%, e a médias da porcentagem de recuperação do padrão variaram de 98 a 101%. O método colori métrico demonstrou ser preciso e exato, podendo ser usado para determinação quantitativa do 17β-estradiol nas formas farmacêuticas estudadas. No método espectrofotométrico na região do UV, o 17β-estradiol foi extraído com etanol absoluto. Determinados os parâmetros analíticos, o método mostrou-se muito simples, rápido, preciso, exato e mais econômico que o método colorimétrico, porém sendo aplicável somente aos comprimidos. / Nowadays, estrogen - based hormonal replacement therapy (HRT) is the treatment of choice for menopause symptoms. For adequate treatment, one of the most widely used estrogens in HRT is 17β-estradiol, which is available in several pharmaceutical dosage forms. A visible spectrophotometric method was developed for quantification of 17β-estradiol in gel, tablets and patches. The method was based on the diazotization reaction of sulfanilamide and azo coupling with 17β-estradiol, forming the brown - reddish azo compound. The reaction behavior was observed for reagent concentration, absorption spectra and color stability. The linearity of the method was observed in a concentration range from 10,0 to 28,0 µg/mL, with correlation coefficient of 0,9996. The detection limit was 0,76 µg/mL and the quantitation limit was 2,30 µg/mL. The standardized method was applied to dosage formulations and simulated samples; the relative standard deviations (RSD) were below of 2% and the average percentage recoveries of standards varied from 98 to 101%. The colorimetric method showed precise, accurate and inexpensive, and can be used for quantitative determination of 17β-estradiol in pharmaceutical formulations studied. On the UV spectrophometric method, the 17β-estradiol was extracted with ethanol absolute. After determining analytical parameters, this method showed very simple, fast, precise, accurate and more economic than colorimetric method but only be applied for tablet formulations.
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