Hyperprolactinemia (hyperPRL) is a highly prevalent adverse side effect in antipsychotic therapy as most antipsychotic drugs are dopamine D2 receptor antagonists. Peony-Glycyrrhiza Decoction (PGD, 芍藥甘草湯) is a classic Chinese herbal formula initially used to treat muscle pain and spasm. Our pilot clinical studies have confirmed the effectiveness of PGD in alleviating antipsychotic-induced hyperPRL in patients with schizophrenia. In the present study, we further examined the effects of PGD, its individual herbal preparations and major compounds in suppressing prolactin (PRL) hyperactivity in in vitro and in vivo models and underlying mechanisms. PGD treatment significantly suppressed PRL secretion in MMQ cells, an exemplary model of hyperPRL that is derived from pituitary adenoma cells. PGD also suppressed PRL synthesis of MMQ cells in a dose-dependent manner; however, these suppressive effects were completely abolished by pretreatment with 10 μM haloperidol, a dopamine D2 receptor antagonist. PGD did not affect hyperactive PRL in GH3 cells that lack the D2 receptor expression, but PGD significantly increased the expressions of the D2 receptor and dopamine transporters (DAT) in PC12 cells. In the rat model of hyperPRL produced by repeated injection with the dopamine blocker metoclopramide (MCP), PGD (5 - 10 g/kg daily) treatment for 14 days significantly reduced elevated serum PRL and the reduced magnitude was similar to that of 0.6 mg/kg bromocriptine (BMT), a dopamine D2 receptor agonist used for treating hyperPRL. Both PGD and BMT did not alter serum estradiol, but PGD reversed MCP-induced decreased serum progesterone to control level, whereas BMT did not. Similar to BMT, PGD treatment displayed a great effect in reversing the MCP-induced reduction of the expressions of D2 receptor, DAT and tyrosine hydroxylase in both the pituitary and the hypothalamus, in particular the arcuate nucleus, but both had least effects on the expressions of PRL in the pituitary and hypothalamus. The anti-hyperPRL effects of individual herbal preparation and major compounds of PGD were further examined in cultured cells. The three herbal preparations, Paeoniae Radix (PR) alone, Glycyrrhiza Radix (GR) alone and the pooled PR and GR individual preparation (PR+GR), and the three major constituents 18β-glycyrrhetinic acid (GA), liquiritigenin (LQ) and paeoniflorin (PF), were tested. All these preparations and constituents displayed significant effects in suppressing PRL hyperactivity and enhancing the expressions of dopamine mediators. However, PR had the most robust anti-hyperPRL effects compared to PGD and other preparations and constituents. The present study provides experimental evidence confirming the clinical effects of PGD in suppressing antipsychotic-associated hyperPRL. Not only D2 receptor is involved in the anti-hyperPRL effect of PGD, it is also associated with the modulation of other dopamine mediators and sex hormones. The finding that the magnitudes of the anti-hyperPRL effects of PGD and of combined PR and GR are similar suggests that boiling the herbs together or or separately shows the same effects. Additionally, PR preparation appears to be more efficacious in reducing hyperPRL compared to GR preparation which deserves to be further investigated. / published_or_final_version / Chinese Medicine / Doctoral / Doctor of Philosophy

Thesis (Ph.D.)--Tufts University, 2001. / Adviser: Shuk-mei Ho. Submitted to the Dept. of Biology. Includes bibliographical references (leaves 178-196). Access restricted to members of the Tufts University community. Also available via the World Wide Web;

Background: Prolactin-secreting tumours( prolactinomas) are the commonest type of pituitary tumour, accounting for approximately 30 to 40 %% of all pituitary adenomas. Although there is ample epidemiologic and clinic data from Industrialised countries there remains sparse data from Africa. Specifically, the clinical presentation, and hormonal deficiencies and treatment outcomes in the South Africa have not been described. Methods: A retrospective study of all patients with a diagnosis of prolactinoma attending the Endocrine and Pituitary Clinics at Groote Schuur Hospital over a 12-month period, between March 2019-March 2020. Patients folders were reviewed to retrieve the following information: demographic data, clinical presentation, clinical signs, prolactinoma phenotype, hormonal deficiencies, treatment modalities and clinical outcomes. Results: Over a 12-month period 52 patients were included in this study, females 73% (n=38), mean age of all participants was 46.1 ± 14.6 years. A macroprolactinoma was present in 67.3% (n=35) of patients and 32.7% (n=17) of patients had a microprolactinoma. In the macroprolactinoma group: the common presenting symptoms were headache 88.6% (n=33), altered vision 40% (n=14) and , in females, amenorrhoea 63.6% (n=14) but a cranial nerve palsy 17.1% (n=6) and apoplexy 5.7% (n=2) were uncommon. . In the microprolactinoma group the common presenting symptoms included amenorrhoea 75% (n=12), galactorrhoea 70.6% (n=12), headache 64.7% (n=11). On presentation the majority of patients with a macroadenoma had at least one hormonal abnormality with hypogonadism 73.1% (n=19) being most common, followed by hypothyroidism 53.8% (n=14) and hypoadrenalism 30% (n=8). Over 50% of patients with a giant adenoma had panhypopituitarism with hypogonadism in 100%, hypothyroidism in 77.8% (n=7) and hypoadrenalism in 66.7% (n=6). Hormonal deficiencies in the microadenoma group on presentation included hypogonadism 64.7% (n=11), hypothyroidism 35.3 (n=6) and one patient had hypoadrenalism. All patients received medical treatment, however, in the macroadenoma group 4 patients required surgical debulking of the tumour, 3 patients required a ventriculo-peritoneal (VP) shunt for hydrocephalus and 2 patients required radiation. After a median follow-up of 46.5 months, the median prolactin level decreased from 322.5 ug/l (94.0-4282.0) at presentation to 17.5 ug/l (8.6-82.5) at follow-up. In parallel there was a reduction of 12.2 ±9.7 mm in tumour size after a mean of 59.8 ±53.3 months. There was resolution of hypogonadism in 56.4% (n=22), of hypothyroidism in 2.7% (n=2) and hypoadrenalism only resolved in 1 patient. Conclusions: Most patients with a prolactinoma are symptomatic and have at least one hormone deficiency on presentation. With medical management most patients experienced a reduction in prolactin levels and tumour size. . This was associated with the resolution of hypogonadism in the majority, however, hypothyroidism and hypoadrenalism are unlikely to resolve despite a reduction in tumour size.

Introdução: A prolactina (PRL) exerce efeitos imunoestimulatórios in vitro e in vivo, porém a literatura é controversa quanto ao papel deste hormônio na atividade do Lúpus Eritematoso Sistêmico (LES). A macroprolactina possui menor atividade biológica in vivo e poderia explicar os resultados díspares. Objetivos: avaliar a prevalência de hiperprolactinemia e macroprolactinemia em pacientes lúpicas; analisar a correlação entre a atividade do LES e PRL, e interferência da macroprolactina nesta associação. Casuística e Métodos: Em 73 mulheres com LES ativo foi dosada a PRL pelo Immulite 2000®, e a macroprolactina pelo método do Polietilenoglicol (momento 1); em 62 destas pacientes foi colhida uma segunda amostra com a menor atividade do LES ao longo do tratamento (momento 2). Os controles foram 29 mulheres hígidas no menacme (grupo C) e 34 gestantes no terceiro trimestre (grupo G). Resultados: Houve 15 casos (20,55%) de hiperprolactinemia nas lúpicas, e nenhum entre as mulheres hígidas (p = 0,005). Todas as gestantes apresentaram hiperprolactinemia. A concentração de PRL foi maior (Med = 11,70 ng/ml) (p = 0,01) no LES do que no grupo C (Med = 8,81ng/ml), e correlacionou-se com a atividade da doença pelo SLEDAI (r = 0,41; p = 0,0003) no momento 1. No LES muito ativo os níveis de PRL foram maiores do que na doença inativa (Med = 17,10 ng/ml vs. Med = 8,36 ng/ml) (p< 0,01), e moderadamente ativa (Med = 7,75 ng/ml) (p < 0,05). Dentre as lúpicas hiperprolactinêmicas, 04 casos (26,7%) foram devidos à macroprolactina, e nas gestantes, 02 casos (5,9%). O LES foi tão ativo na macroprolactinemia quanto nos casos pela forma monomérica, porém a correlação entre PRL e SLEDAI foi maior para a PRL livre (r = 0,44; p = 0,0001). O tratamento das pacientes lúpicas hiperprolactinêmicas resultou em diminuição da concentração de PRL (Me momento 1 = 56,71 ± 43,87 ng/ml vs. Me momento 2 = 18,68 ± 24,20 ng/ml) (p = 0,015). Conclusões: pacientes lúpicas apresentam hiperprolactinemia mais frequentemente do que mulheres hígidas, e a PRL correlaciona-se com a atividade do LES. A macroprolactinemia não é marcador de doença inativa/pouco ativa. / Introduction: Prolactin (PRL) is a hormone with widespread influences in the cells of the immune system, which have been demonstrated by several in vitro and in vivo studies. However, the role of this hormone in the pathogenesis of Systemic Lupus Erythematosus (SLE) is controversial within the medical literature. The potentially lower biological activity of macroprolactin could explain the disparity of the results. Methods: PRL levels were determined by chemo luminescence method (Immulite 2000®) in 73 women with active SLE (group L), while the screening for macroprolactinemia was determined by the polyethylene glycol precipitation method (first moment). Sixty two of these patients had their PRL levels determined in a second occasion, when the disease was inactive or with the lowest activity observed after treatment (second moment). The control groups were 29 healthy women (group C) and 34 third-trimester healthy pregnant (group P). The levels of PRL were correlated with the SLE Disease Activity Index (SLEDAI). Results: In the study group there were 15 (20.55%) cases of hyperprolactinemia, while in the group C there were none (p = 0,005). All pregnant women presented hyperprolactinemia. Prolactin levels were higher in group L (Med = 11,70 ng/ml) then in group C (Med = 8,81ng/ml) (p = 0,01) and correlated with the SLEDAI in the first moment (r = 0,41; p = 0,0003). We also detected that PRL levels were higher at highly active SLE (SLEDAI ¡Ý 11) than when the disease was inactive (SLEDAI = 0) (Med = 17,10 ng/ml vs. Med = 8,36 ng/ml) (p< 0,01) or moderately active SLE (6 ¡Ü SLEDAI ¡Ü 10) (Med = 7,75 ng/ml) (p<0,05). In the 15 patients of group L with hyperprolactinemia, there were 04 cases of macroprolactinemia (26.7%), while 02 subjects in group P presented it (5.9%). SLE was as active in the patients with hyperprolactinemia caused by the monomeric form of the hormone, as in the ones with macroprolactinemia. The correlation of the PRL levels and the SLEDAI was, nevertheless, stronger for free PRL (r = 0,44; p = 0,0001). The SLE treatment in the hyperprolactinemic patients reduced PRL levels from 56,71 ng/ml (sd = 43,87) to 18,68 ng/ml (± 24,20) (p = 0,015). Discussion: the frequency of hyperprolactinemia is higher in SLE than in the general population, and the levels of PRL correlate with the activity of the disease. Macroprolactin is also associated to active SLE.

Introdução: A prolactina (PRL) exerce efeitos imunoestimulatórios in vitro e in vivo, porém a literatura é controversa quanto ao papel deste hormônio na atividade do Lúpus Eritematoso Sistêmico (LES). A macroprolactina possui menor atividade biológica in vivo e poderia explicar os resultados díspares. Objetivos: avaliar a prevalência de hiperprolactinemia e macroprolactinemia em pacientes lúpicas; analisar a correlação entre a atividade do LES e PRL, e interferência da macroprolactina nesta associação. Casuística e Métodos: Em 73 mulheres com LES ativo foi dosada a PRL pelo Immulite 2000®, e a macroprolactina pelo método do Polietilenoglicol (momento 1); em 62 destas pacientes foi colhida uma segunda amostra com a menor atividade do LES ao longo do tratamento (momento 2). Os controles foram 29 mulheres hígidas no menacme (grupo C) e 34 gestantes no terceiro trimestre (grupo G). Resultados: Houve 15 casos (20,55%) de hiperprolactinemia nas lúpicas, e nenhum entre as mulheres hígidas (p = 0,005). Todas as gestantes apresentaram hiperprolactinemia. A concentração de PRL foi maior (Med = 11,70 ng/ml) (p = 0,01) no LES do que no grupo C (Med = 8,81ng/ml), e correlacionou-se com a atividade da doença pelo SLEDAI (r = 0,41; p = 0,0003) no momento 1. No LES muito ativo os níveis de PRL foram maiores do que na doença inativa (Med = 17,10 ng/ml vs. Med = 8,36 ng/ml) (p< 0,01), e moderadamente ativa (Med = 7,75 ng/ml) (p < 0,05). Dentre as lúpicas hiperprolactinêmicas, 04 casos (26,7%) foram devidos à macroprolactina, e nas gestantes, 02 casos (5,9%). O LES foi tão ativo na macroprolactinemia quanto nos casos pela forma monomérica, porém a correlação entre PRL e SLEDAI foi maior para a PRL livre (r = 0,44; p = 0,0001). O tratamento das pacientes lúpicas hiperprolactinêmicas resultou em diminuição da concentração de PRL (Me momento 1 = 56,71 ± 43,87 ng/ml vs. Me momento 2 = 18,68 ± 24,20 ng/ml) (p = 0,015). Conclusões: pacientes lúpicas apresentam hiperprolactinemia mais frequentemente do que mulheres hígidas, e a PRL correlaciona-se com a atividade do LES. A macroprolactinemia não é marcador de doença inativa/pouco ativa. / Introduction: Prolactin (PRL) is a hormone with widespread influences in the cells of the immune system, which have been demonstrated by several in vitro and in vivo studies. However, the role of this hormone in the pathogenesis of Systemic Lupus Erythematosus (SLE) is controversial within the medical literature. The potentially lower biological activity of macroprolactin could explain the disparity of the results. Methods: PRL levels were determined by chemo luminescence method (Immulite 2000®) in 73 women with active SLE (group L), while the screening for macroprolactinemia was determined by the polyethylene glycol precipitation method (first moment). Sixty two of these patients had their PRL levels determined in a second occasion, when the disease was inactive or with the lowest activity observed after treatment (second moment). The control groups were 29 healthy women (group C) and 34 third-trimester healthy pregnant (group P). The levels of PRL were correlated with the SLE Disease Activity Index (SLEDAI). Results: In the study group there were 15 (20.55%) cases of hyperprolactinemia, while in the group C there were none (p = 0,005). All pregnant women presented hyperprolactinemia. Prolactin levels were higher in group L (Med = 11,70 ng/ml) then in group C (Med = 8,81ng/ml) (p = 0,01) and correlated with the SLEDAI in the first moment (r = 0,41; p = 0,0003). We also detected that PRL levels were higher at highly active SLE (SLEDAI ¡Ý 11) than when the disease was inactive (SLEDAI = 0) (Med = 17,10 ng/ml vs. Med = 8,36 ng/ml) (p< 0,01) or moderately active SLE (6 ¡Ü SLEDAI ¡Ü 10) (Med = 7,75 ng/ml) (p<0,05). In the 15 patients of group L with hyperprolactinemia, there were 04 cases of macroprolactinemia (26.7%), while 02 subjects in group P presented it (5.9%). SLE was as active in the patients with hyperprolactinemia caused by the monomeric form of the hormone, as in the ones with macroprolactinemia. The correlation of the PRL levels and the SLEDAI was, nevertheless, stronger for free PRL (r = 0,44; p = 0,0001). The SLE treatment in the hyperprolactinemic patients reduced PRL levels from 56,71 ng/ml (sd = 43,87) to 18,68 ng/ml (± 24,20) (p = 0,015). Discussion: the frequency of hyperprolactinemia is higher in SLE than in the general population, and the levels of PRL correlate with the activity of the disease. Macroprolactin is also associated to active SLE.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2016-04-06T14:30:58Z No. of bitstreams: 2 Tese - Eliane Gouveia de Morais Sanchez - 2015.pdf: 1194460 bytes, checksum: 67eee577cf08aa1bdca9f0cbdddad7f1 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-04-06T14:33:14Z (GMT) No. of bitstreams: 2 Tese - Eliane Gouveia de Morais Sanchez - 2015.pdf: 1194460 bytes, checksum: 67eee577cf08aa1bdca9f0cbdddad7f1 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2016-04-06T14:33:14Z (GMT). No. of bitstreams: 2 Tese - Eliane Gouveia de Morais Sanchez - 2015.pdf: 1194460 bytes, checksum: 67eee577cf08aa1bdca9f0cbdddad7f1 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-10-29 / Infertility reflects the inability of a couple to achieve pregnancy after one year of sexual intercourse without contraception. Most common causes are related to ovulatory disorders such as hyperprolactinaemia, polycystic ovary syndrome, thyroid dysfunction, corpus luteum deficiency, among others, and can be identified mainly by ultrasound and hormonal dosage. The overall objective of this study was to evaluate the influence of prolactin ovulation in infertile women suffering from regular cycles. It is case-control study, whose sample consisted of 343 women with age range 20-40 years old, attended from 2000 to 2014 in LabRep-HC / UFG and an office of Obstetrics and Gynecology, private network in Goiânia, Goiás, Brazil. Data collection was performed by analyzing the physical records stored in Medical Records and Health Information Service (SAMIS) and electronics made available by the database (Sisfert) (© Approbato, 2013). The patients were classified according to the ovulation state measured by progestogen dosage (≥ 5.65 ng / ml and 5.65 - 9.9 ng / ml) and ovulation by monitoring the ultrasound being divided into four groups: (I ) probable ovulation, (II) likely anovulation. In Groups I and II were compared with the percentages of patients who do not ovulate with normal prolactin (3 to 20 ng / ml) versus moderately elevated prolactin (21 to 29 ng / ml). In group III were evaluated Normal progesterone levels (≥ 10 ng / mL) versus low progesterone (Group IV) (5.65 - 9.9 ng / ml) was considered as LUF (non-luteinized ruptured follicle). The groups were comparable for comparable as to age, body mass index (BMI), duration of infertility, FSH (ng / ml) TSH (mIU / l), LH (IU / l) and oestradiol (ng / dL) . SPSS Statistics 20.0 software and Bioestat (version 5.3) were used to for data analysis and chi-square test (X2) to assess differences between proportions. Where it is not for statistical analysis were calculated mean and standard deviation of the variables under study. The results demonstrated that moderate elevation of prolactin (21-29 ng / ml) caused a significant reduction (p = 0.03) in the ovulation rate of infertile patients with regular cycles considering as a criterion for ovulation progesterone levels ≥5, 65 ng / ml. When evaluated the influence of low progesterone on ovulation monitored by ultrasound was observed that these levels can significantly reduce the percentage of ovulation. It is concluded that a moderate hyperprolactinaemia and low progesterone can negatively influence the regulation of ovulation in infertile women with regular menstrual cycles. / A infertilidade reflete a incapacidade de um casal conseguir gravidez após um ano de relações sexuais sem contracepção. Causas mais comuns estão relacionadas a disfunções ovulatórias como a hiperprolactinemia, síndrome de ovário policístico, disfunções da tireóide, deficiência de corpo lúteo, entre outras, e que podem ser identificadas, principalmente, por dosagem hormonal e ultrassonografia. O objetivo geral desse estudo foi avaliar a influência dos níveis de prolactina na ovulação de mulheres inférteis portadoras de ciclos regulares. Trata-se de estudo caso-controle, cuja amostra foi composta por 343 mulheres com faixa etária compreendida de 20 a 40 anos, atendidas no período de 2000 a 2014 no LabRep-HC/UFG e em um consultório de Ginecologia e Obstetrícia da rede particular em Goiânia, Goiás, Brasil. A coleta de dados foi feita pela análise dos prontuários físicos armazenados no Serviço de Arquivo Médico e Informações em Saúde (SAMIS) e eletrônicos disponibilizados pelo banco de dados (Sisfert) (©Approbato, 2013). As pacientes foram classificadas de acordo com o estado ovulatório avaliado pela dosagem de progesterona (≥ 5,65 ng/ml e de 5,65 – 9,9 ng/ml) e monitorização da ovulação pelo ultrassom sendo divididas em quatro grupos: (I) provável ovulação, (II) provável anovulação. Nos grupos I e II foram comparadas as porcentagens de pacientes que não ovulavam com prolactina normal (3 a 20 ng/ml) versus prolactina moderadamente elevada (21 a 29 ng/ml). No grupo III foram avaliados os níveis de progesterona normal (≥ 10 ng/ml) versus progesterona baixa (Grupo IV) (5,65 – 9,9 ng/ml) considerada como LUF (Folículo Luteinizado não-roto). Os grupos foram pareados para a comparabilidade quanto a idade, índice de massa corporal (IMC), duração da infertilidade, FSH (ng/ml), TSH (mUI/l), LH (UI/l) e estradiol (ng/dl). Os programas SPSS Statistics 20.0 e Bioestat (versão 5.3) foram utilizados para para a análise dos dados e o teste Qui quadrado (X2) para avaliar as diferenças entre proporções. Onde não coube análise estatística foram calculadas média e desvio padrão das variáveis em estudo. Os resultados demonstraram que a elevação moderada da prolactina (21-29 ng/ml) provocou a redução significativa (p=0,03) na porcentagem de ovulação das pacientes inférteis portadoras de ciclos regulares considerando como critério de ovulação níveis de progesterona ≥5,65 ng/ml. Quando avaliada a influência da progesterona baixa sobre a ovulação monitorada pelo ultrassom foi observado que esses níveis podem reduzir de forma significativa a porcentagem de ovulação. Conclui-se que a hiperprolactinemia moderada e a progesterona baixa podem influenciar negativamente na regulação da ovulação de mulheres inférteis com ciclos regulares.

INTRODUÇÃO: A prolactina é um hormônio polipeptídico, que possui reconhecida ação sistêmica, principalmente na fisiologia da reprodução, porém, seu desequilíbrio, em especial a hiperprolactinemia, é cada vez mais frequente na prática clínica. Apesar de ser um distúrbio relativamente comum, ainda existem dúvidas quanto aos efeitos moleculares da hiperprolactinemia no trato genital, particularmente no útero, e também na hipófise. O presente estudo teve por objetivo verificar os efeitos da hiperprolactinemia induzida pela metoclopramida na expressão gênica da prolactina e de seus receptores no útero e na hipófise de camundongo fêmea. MÉTODOS: Utilizaram-se 49 camundongos fêmeas (Wistar), randomicamente divididas em 7 grupos contendo 7 animais cada: 1) SS não ovariectomizadas que receberam solução salina (veículo); 2) M não ovariectomizadas tratadas com metoclopramida; 3) OSS ovariectomizadas tratadas com solução salina (veículo); 4) OM ovariectomizadas tratadas com metoclopramida; 5) OME ovariectomizadas tratadas com metoclopramida e 17-estradiol; 6) OMP ovariectomizadas tratadas com metoclopramida e progesterona micronizada; 7) OMEP ovariectomizadas tratadas com metoclopramida, 17-estradiol e progesterona micronizada. Após 50 dias os animais foram sacrificados sendo retirados o útero e a hipófise de cada animal para extração do ácido ribonucleico total, que foi utilizado para a síntese de ácido desoxirribonucleico complementar e avaliação da expressão gênica da prolactina e das diferentes isoformas de seus receptores, por reação em cadeia da polimerase em tempo real. RESULTADOS: Na hipófise, em animais não ovariectomizados, o tratamento com metoclopramida aumentou a expressão do gene que codifica a prolactina em relação ao tratamento apenas com o veículo. Nos animais castrados, a progesterona isoladamente ou associada ao estrogênio determinou o incremento do RNA mensageiro da prolactina em relação aos outros animais castrados que receberam outras combinações de tratamento. Este efeito foi semelhante ao da metoclopramida em animais com os ovários intactos. Em relação ao receptor de prolactina, o estrogênio e a progesterona, isoladamente, foram responsáveis pelo incremento da isoforma S2. No útero houve aumento na expressão de RNA mensageiro de prolactina após tratamento com metoclopramida ou com tratamento isolado ou combinado de estrogênio e progesterona. A ovariectomia determinou a redução da expressão das isoformas S1 e S2 do receptor de prolactina de todas as isoformas estudadas. Já o tratamento estroprogestativo determinou elevação da formas S3 e L do receptor, enquanto com a progesterona isoladamente causou apenas o incremento da forma L do receptor da prolactina no útero dos animais castrados. CONCLUSÕES: Nossos dados sugerem que o tratamento com metoclopramida altera de forma diferente a expressão de prolactina e de seus receptores quando se compara o resultado da hipófise em relação ao útero em camundongos fêmeas castrados e tratados com esteróides sexuais / INTRODUCTION: Prolactin is a polypeptide hormone with a recognized systemic action mainly on reproductive physiology. However, prolactin imbalance, particularly hyperprolactinemia, is increasingly more frequent in clinical practice. Although it is a comparatively common disorder, there are still doubts about the molecular effects of hyperprolactinemia on the genital tract especially in the uterus and the pituitary. The present study aimed at verifying the effects of metoclopramide-induced hyperprolactinemia on the gene expression of prolactin and its receptors in the uterus and pituitary of the female mouse. METHODS: Forty-nine female Wistar mice were randomized to 7 equal-sized groups as follows: 1) SS nonoophorectomized mice treated with saline solution (vehicle); 2) M nonoophorectomized mice treated with metoclopramide; 3) OSS oophorectomized mice treated with saline solution (vehicle); 4) OM oophorectomized mice treated with metoclopramide; 5) OME oophorectomized mice treated with metoclopramide and 17-estradiol; 6) OMP oophorectomized mice treated with metoclopramide and micronized progesterone; 7) OMEP oophorectomized mice treated with metoclopramide, 17-estradiol, and micronized progesterone. The animals were sacrificed 50 days after the end of the treatment, and the uterus and pituitary of each animal were removed for extraction of total ribonucleic acid, which was then used for synthesizing complementary deoxyribonucleic acid and for evaluating the gene expression of prolactin and the different isoforms of its receptors by the real-time polymerase chain reaction. RESULTS: In the pituitary of the nonoophorectomized mice, the treatment with metoclopramide against that with vehicle alone increased the expression of the prolactin-encoding gene. In the castrated animals, progesterone by itself or in conjunction with estrogen determined a raise in prolactin messenger RNA as opposed to the two other treatments with different combinations. This effect was similar to that produced by metoclopramide in animals with intact ovaries. Estrogen and progesterone, acting independently of each other, were responsible for the increase in the S2 isoform of the prolactin receptor. In the uterus, there was heightened expression of prolactin messenger RNA under the effect of the treatment with metoclopramide or with estrogen and/or progesterone. Oophorectomy caused a greater reduction in expression of the prolactin receptor S1 and S2 isoforms than in the other isoforms. However, the combined estrogen plus progesterone treatment led to an increase in the S3 and L forms of the receptor, while progesterone alone resulted solely in a higher expression of the L form of the prolactin receptor in the endometrium of the castrated mice. CONCLUSION: Our data suggest that metoclopramide treatment induces different changes in the expression of prolactin and its receptors according to whether the effect occurs in the pituitary or the uterus of castrated female mice treated with sex steroids

A macroprolactinemia é condição freqüente na hiperprolactinemia e em geral, sem impacto clínico. Os dados sobre a atividade biológica da macroprolactina (bbPRL) são controversos e baseados em bioensaio heterólogo com células de rato Nb2. A atividade biológica da bbPRL é observada in vitro e não in vivo, provavelmente porque seu alto peso molecular evita sua passagem pelos capilares. A bioatividade da bbPRL talvez varie de acordo com a especificidade do receptor de prolactina (PRLR). Avaliamos a bioatividade da bbPRL de indivíduos macroprolactinêmicos (Grupo I, n = 18) e da PRL monomérica (mPRL) de pacientes hiperprolactinêmicos sem bbPRL (Grupo II, n = 5) em Nb2 e em células Ba/F-LLP, transfectadas com o PRLR humano. Enquanto ambos ensaios apresentam resultados similares para a atividade de mPRL, nossos resultados indicam que a atividade da bbPRL é presente em ensaio heterólogo e não em ensaio homólogo. O ensaio Ba/F-LLP é sensível e apresenta melhor correlação com a atividade in vivo da bbPRL / Macroprolactinemia is a frequent finding in hyperprolactinemic individuals, usually without clinical impact. Data on biological activity of macroprolactin (bbPRL) is mostly based on a heterologous bioassay (Nb2 cell). Biological activity of bbPRL observed in vitro but not in vivo maybe due to its high molecular weight preventing its passage through capillary barrier. Alternatively, bbPRL bioactivity may differ depending on the PRL receptor species specificity. BbPRL from macroprolactinemic individuals and monomeric PRL (mPRL) from hyperprolactinemic patients without macroprolactinemia were tested in two bioassays: Nb2 and in Ba/F-LLP, which expresses human prolactin receptor. While both bioassays achieve similar results considering mPRL activity, our results indicate that bbPRL displays activity in a heterologous but not in a homologous bioassay, consistently with the apparent absence of bbPRL bioactivity in vivo

INTRODUÇÃO: A prolactina é um hormônio polipeptídico, que possui reconhecida ação sistêmica, principalmente na fisiologia da reprodução, porém, seu desequilíbrio, em especial a hiperprolactinemia, é cada vez mais frequente na prática clínica. Apesar de ser um distúrbio relativamente comum, ainda existem dúvidas quanto aos efeitos moleculares da hiperprolactinemia no trato genital, particularmente no útero, e também na hipófise. O presente estudo teve por objetivo verificar os efeitos da hiperprolactinemia induzida pela metoclopramida na expressão gênica da prolactina e de seus receptores no útero e na hipófise de camundongo fêmea. MÉTODOS: Utilizaram-se 49 camundongos fêmeas (Wistar), randomicamente divididas em 7 grupos contendo 7 animais cada: 1) SS não ovariectomizadas que receberam solução salina (veículo); 2) M não ovariectomizadas tratadas com metoclopramida; 3) OSS ovariectomizadas tratadas com solução salina (veículo); 4) OM ovariectomizadas tratadas com metoclopramida; 5) OME ovariectomizadas tratadas com metoclopramida e 17-estradiol; 6) OMP ovariectomizadas tratadas com metoclopramida e progesterona micronizada; 7) OMEP ovariectomizadas tratadas com metoclopramida, 17-estradiol e progesterona micronizada. Após 50 dias os animais foram sacrificados sendo retirados o útero e a hipófise de cada animal para extração do ácido ribonucleico total, que foi utilizado para a síntese de ácido desoxirribonucleico complementar e avaliação da expressão gênica da prolactina e das diferentes isoformas de seus receptores, por reação em cadeia da polimerase em tempo real. RESULTADOS: Na hipófise, em animais não ovariectomizados, o tratamento com metoclopramida aumentou a expressão do gene que codifica a prolactina em relação ao tratamento apenas com o veículo. Nos animais castrados, a progesterona isoladamente ou associada ao estrogênio determinou o incremento do RNA mensageiro da prolactina em relação aos outros animais castrados que receberam outras combinações de tratamento. Este efeito foi semelhante ao da metoclopramida em animais com os ovários intactos. Em relação ao receptor de prolactina, o estrogênio e a progesterona, isoladamente, foram responsáveis pelo incremento da isoforma S2. No útero houve aumento na expressão de RNA mensageiro de prolactina após tratamento com metoclopramida ou com tratamento isolado ou combinado de estrogênio e progesterona. A ovariectomia determinou a redução da expressão das isoformas S1 e S2 do receptor de prolactina de todas as isoformas estudadas. Já o tratamento estroprogestativo determinou elevação da formas S3 e L do receptor, enquanto com a progesterona isoladamente causou apenas o incremento da forma L do receptor da prolactina no útero dos animais castrados. CONCLUSÕES: Nossos dados sugerem que o tratamento com metoclopramida altera de forma diferente a expressão de prolactina e de seus receptores quando se compara o resultado da hipófise em relação ao útero em camundongos fêmeas castrados e tratados com esteróides sexuais / INTRODUCTION: Prolactin is a polypeptide hormone with a recognized systemic action mainly on reproductive physiology. However, prolactin imbalance, particularly hyperprolactinemia, is increasingly more frequent in clinical practice. Although it is a comparatively common disorder, there are still doubts about the molecular effects of hyperprolactinemia on the genital tract especially in the uterus and the pituitary. The present study aimed at verifying the effects of metoclopramide-induced hyperprolactinemia on the gene expression of prolactin and its receptors in the uterus and pituitary of the female mouse. METHODS: Forty-nine female Wistar mice were randomized to 7 equal-sized groups as follows: 1) SS nonoophorectomized mice treated with saline solution (vehicle); 2) M nonoophorectomized mice treated with metoclopramide; 3) OSS oophorectomized mice treated with saline solution (vehicle); 4) OM oophorectomized mice treated with metoclopramide; 5) OME oophorectomized mice treated with metoclopramide and 17-estradiol; 6) OMP oophorectomized mice treated with metoclopramide and micronized progesterone; 7) OMEP oophorectomized mice treated with metoclopramide, 17-estradiol, and micronized progesterone. The animals were sacrificed 50 days after the end of the treatment, and the uterus and pituitary of each animal were removed for extraction of total ribonucleic acid, which was then used for synthesizing complementary deoxyribonucleic acid and for evaluating the gene expression of prolactin and the different isoforms of its receptors by the real-time polymerase chain reaction. RESULTS: In the pituitary of the nonoophorectomized mice, the treatment with metoclopramide against that with vehicle alone increased the expression of the prolactin-encoding gene. In the castrated animals, progesterone by itself or in conjunction with estrogen determined a raise in prolactin messenger RNA as opposed to the two other treatments with different combinations. This effect was similar to that produced by metoclopramide in animals with intact ovaries. Estrogen and progesterone, acting independently of each other, were responsible for the increase in the S2 isoform of the prolactin receptor. In the uterus, there was heightened expression of prolactin messenger RNA under the effect of the treatment with metoclopramide or with estrogen and/or progesterone. Oophorectomy caused a greater reduction in expression of the prolactin receptor S1 and S2 isoforms than in the other isoforms. However, the combined estrogen plus progesterone treatment led to an increase in the S3 and L forms of the receptor, while progesterone alone resulted solely in a higher expression of the L form of the prolactin receptor in the endometrium of the castrated mice. CONCLUSION: Our data suggest that metoclopramide treatment induces different changes in the expression of prolactin and its receptors according to whether the effect occurs in the pituitary or the uterus of castrated female mice treated with sex steroids

A macroprolactinemia é condição freqüente na hiperprolactinemia e em geral, sem impacto clínico. Os dados sobre a atividade biológica da macroprolactina (bbPRL) são controversos e baseados em bioensaio heterólogo com células de rato Nb2. A atividade biológica da bbPRL é observada in vitro e não in vivo, provavelmente porque seu alto peso molecular evita sua passagem pelos capilares. A bioatividade da bbPRL talvez varie de acordo com a especificidade do receptor de prolactina (PRLR). Avaliamos a bioatividade da bbPRL de indivíduos macroprolactinêmicos (Grupo I, n = 18) e da PRL monomérica (mPRL) de pacientes hiperprolactinêmicos sem bbPRL (Grupo II, n = 5) em Nb2 e em células Ba/F-LLP, transfectadas com o PRLR humano. Enquanto ambos ensaios apresentam resultados similares para a atividade de mPRL, nossos resultados indicam que a atividade da bbPRL é presente em ensaio heterólogo e não em ensaio homólogo. O ensaio Ba/F-LLP é sensível e apresenta melhor correlação com a atividade in vivo da bbPRL / Macroprolactinemia is a frequent finding in hyperprolactinemic individuals, usually without clinical impact. Data on biological activity of macroprolactin (bbPRL) is mostly based on a heterologous bioassay (Nb2 cell). Biological activity of bbPRL observed in vitro but not in vivo maybe due to its high molecular weight preventing its passage through capillary barrier. Alternatively, bbPRL bioactivity may differ depending on the PRL receptor species specificity. BbPRL from macroprolactinemic individuals and monomeric PRL (mPRL) from hyperprolactinemic patients without macroprolactinemia were tested in two bioassays: Nb2 and in Ba/F-LLP, which expresses human prolactin receptor. While both bioassays achieve similar results considering mPRL activity, our results indicate that bbPRL displays activity in a heterologous but not in a homologous bioassay, consistently with the apparent absence of bbPRL bioactivity in vivo