Treatment with Raloxifene Induces the Expression of Kisspeptin, Insulin, and Androgen Receptors in Bones of Castrated Adult Female Rats / Tratamento com raloxifeno induz à expressão dos receptores de kisspeptina, insulina e androgênio em ossos de ratas adultas castradas

Abstract Objective To evaluate the effects of estrogen, raloxifene and genistein on the expression of KISS1 (kisspeptin), KISS1R (kisspeptin receptor), AR (androgen receptor) and INSR (insulin receptor) in the bones of ovariectomized rats. Methods Forty-eight adult rats were randomly divided into 6 groups, containing 8 animals each: G1-nonovariectomized control; G2-ovariectomized and treated with conjugated equine estrogens (50 µg/Kg/day); G3-ovariectomized and treated with raloxifene (0.75 mg/kg/day); G4-ovariectomized animal that received soy extract with genistein (300 mg/kg/day); G5-ovariectomized animal that received estrogen and genistein; and G6-ovariectomized animal that received estrogen and raloxifene. Three months after surgery, the castrated animals received the drugs orally daily for 120 days. All animals were sacrificed after this period, by deepening the anesthesia. The left tibia was removed for total RNA extraction and analysis of gene expression of KISS1, KISS1R, AR and INSR, by quantitative real-time polymerase chain reaction (qRT-PCR). Results KISS1 was not detected in any of the treated groups. KISS1R, INSR and AR showed higher expression in the G3 group (p < 0.001), while lower levels of transcripts for these genes were observed in G4 and G5. G2 animals showed hypoexpression of the evaluated genes. Conclusion The results indicate that raloxifene, alone or combined with estrogen, was able to induce the expression of genes associated with the recovery of bone tissue homeostasis in ovariectomized rats.

Resumo Objetivo Avaliar os efeitos do estrogênio, raloxifeno e genisteína na expressão de KISS1 (kisspeptina), KISS1R (receptor da kisspeptina), AR (receptor de androgênio) e INSR (receptor de insulina) nos ossos de ratas ovariectomizadas. Métodos Quarenta e oito ratas adultas foram divididas aleatoriamente em 6 grupos, contendo 8 animais cada: G1-controle não ovariectomizado); G2-ovariectomizado e tratado com estrogênios conjugados equinos (50 µg/Kg/dia); G3-ovariectomizado e tratado com raloxifeno (0,75 mg/kg/dia); G4-ovariectomizado que recebeu extrato de soja com genisteína (300 mg/kg/dia); G5-ovariectomizado que recebeu estrogênio e genisteína; e G6-ovariectomizado que recebeu estrogênio e raloxifeno. Após 3 meses da cirurgia, os animais castrados receberam os fármacos diariamente por via oral, durante 120 dias. Todos os animais foram sacrificados após esse período, por aprofundamento da anestesia. A tíbia esquerda foi removida para extração de RNA total e análise da expressão gênica de KISS1, KISS1R, AR e INSR, por reação de cadeia de polimerase quantitativa em tempo real (quantitative real-time polymerase chain reaction, qRT-PCR, em inglês). Resultados KISS1 não foi detectado em nenhum dos grupos tratados. KISS1R, INSR e AR mostraram maior expressão no grupo G3 (p < 0,001), enquanto menores níveis de transcritos para esses genes foram observados em G4 e G5. Os animais de G2 apresentaram hipoexpressão dos genes avaliados. Conclusão Os resultados indicam que o raloxifeno, isolado ou combinado com estrogênio, foi capaz de induzir a expressão de genes associados à recuperação da homeostase do tecido ósseo em ratas ovariectomizadas.

Thermosensitive Copolymeric PLGA-PEG-PLGA Nanomicelles of Raloxifene: Synthesis, Formulation, and In-vitro Characterization

The objective of the present research study has been to synthesize (PLGA-PEG-PLGA), a thermolabile and biodegradable triblock copolymer. The polymer was subjected to differential evaluation using NMR, XRD, and FTIR techniques for characterization. Raloxifene, a potential therapeutic for breast cancer, exerts poor water solubility and a low fraction of bioavailability owing to its pharmacokinetic properties. The synthesized polymer was used to load raloxifene-encapsulating polymeric micelles, which were then subjected to several evaluations with the aim of improving solubility and bioavailability standards. Zeta potential for the developed formulation was approximately -0.73 mV, indicating a relatively neutral surface charge. It subsequently emerged that the sizes of the blank and raloxifene-loaded micelles were 40.18 and 42.18 nm, respectively. The drug encapsulating capacity and% drug loading capacity were determined to be 73.4 ± 0.34% and drug loading 6.04 ± 0.002% (w/w), respectively. In-vitro study illustrated a 94.37% sustained release drug profile for 120 hours. In this research study, polymeric raloxifene nanomicelles were successfully prepared to enhance solubility, bioavailability, and antitumor effect due to the smaller size of the micelles.

Effects of estrogen and raloxifene on synaptic density in the hippocampal CA1 region of ovariectomized rats

Abstract Introduction The CA1 region of the hippocampus has an important role in learning and memory. It has been shown that estrogen deficiency may reduce the synaptic density in the region and that hormone replacement therapy may attenuate the reduction. Objectives This study aimed to evaluate the effects of estrogen and raloxifene on the synaptic density profile in the CA1 region of the hippocampus in ovariectomized rats. Methods Sixty ovariectomized three-month-old virgin rats were randomized into six groups (n = 10). Treatments started either three days (early treatment) or sixty days (late treatment) after ovariectomy. The groups received propylene glycol vehicle (0.5 mL/animal/day), equine conjugated estrogens (50 μg/animal/day), or raloxifene (3 mg/kg/day) either early or late after ovariectomy. The drugs were administered orally by gavage for 30 days. At the end of the treatments, the animals were anesthetized and transcardially perfused with ether and saline solution. The brains were removed and prepared for analysis under transmission electron microscopy and later fixed. Results Results showed a significant increase in the synaptic density profile of the hippocampal CA1 region in both the early estrogen (0.534 ± 0.026 µ/m2) and the early raloxifene (0.437 ± 0.012 µ/m2) treatment groups compared to the early or late vehicle-treated control groups (0.338 ± 0.038 µ/m2 and 0.277 ± 0.015 µ/m2 respectively). Conclusions The present data suggest that the raloxifene effect may be lower than that of estrogen, even early or late treatment, on synaptic density in the hippocampus.

Efeito do BioGran® funcionalizado ao raloxifeno através do processamento ultrassônico no reparo ósseo em defeitos críticos de calvárias de ratos / Effect of BioGran® functionalized to Raloxifene through ultrasonic processing in bone repair in critical defects in rat calvaria

Este estudo teve como objetivo analisar o processo de formação óssea, bem como a microarquitetura óssea promovido pela associação entre o BioGran® e diferentes concentrações de raloxifeno através do método da sonoquímica em defeitos críticos realizados em calvária de rato. Em um total de doze ratos machos, um defeito crítico de 5 mm de diâmetro foi feito e preenchido com BioGran® 100% (Bg), BioGran® 90% associado à Raloxifeno 10%, e BioGran® 80% associado à Raloxifeno 20%. Aos 14 e 24 dias pós-operatório, foram aplicados os fluorocromos calceína e alizarina, de modo respectivo. A eutanásia ocorreu aos 30 dias após a realização do procedimento cirúrgico para enxertia dos biomateriais. Análise de microtomografia computadorizada (micro-CT) através dos parâmetros de superfiície de intersecção (i.S), fração de volume ósseo (BV/TV) e densidade de conectividade (Conn.Dn) e por microscopia confocal a laser por meio da dinâmica óssea, superfície de mineralização ativa e a taxa de aposição mineral (MAR). Os dados foram analisados através de análise estatística utilizando o teste de normalidade Shapiro-Wilk e o pós teste de Turkey (p<0.05). Para os parâmetros de micro-CT avaliados os menores valores foram encontrados no grupo BG+RL10% (p<0,05), valores similares foram encontrados entre os grupos BG e BG+RL20%. A microscopia confocal evidenciou melhor mineralização óssea e maior taxa de aposição mineral (MAR) para o Grupo BG+RL20% (p<0,05%). Conclusão: A concentração de Raloxifeno a 20% combinado ao BioGran® pelo método da sonoquímica parece ter acelerado o reparo ósseo(AU)

This study aimed to analyze the bone formation process, as well as the bone microarchitecture promoted by the association between BioGran® and different concentrations of raloxifene through the sonochemistry method in critical defects performed in rat calvaria. In a total of twelve male rats, a critical defect of 5 mm in diameter was made and filled with BioGran® 100% (Bg), BioGran® 90% associated with Raloxifene 10%, and BioGran® 80% associated with Raloxifene 20%. At 14 and 24 days postoperatively, the fluorochromes calcein and alizarin were applied, respectively. Euthanasia occurred 30 days after the surgical procedure for grafting biomaterials. Analysis of computed microtomography (microCT) through the parameters of intersection surface (iS), bone volume fraction (BV / TV) and connectivity density (Conn.Dn) and by confocal microscopy through bone dynamics, surface of active mineralization and the mineral apposition rate (MAR). Data were analyzed through statistical using the Shapiro-Wilk normality test and the Turkey post-test (p <0.05). For the me microCT parameters evaluated, the lowest values were found in the BG + RL10% group (p<0.05), similar values were found between the BG and BG + RL20% groups. Confocal microscopy showed better bone mineralization and higher mineral apposition rate (MAR) for the BG + RL20% Group (p < 0.05%). Conclusion: The concentration of Raloxifene at 20% combined with BioGran® by the sonochemistry method seems to have accelerated bone repair(AU)

Análise do reparo ósseo da superfície de implante revestida com raloxifeno através das análises topográfica, biomecânica, histológica, histométrica e imunoistoquímica / Analysis of bone repair of the implant surface coated with raloxifene through topographic, biomechanical, histological, histometric and immunohistochemical analyzes

Introdução: A fim de melhorar a osseointegração, pesquisas tem buscado tratamentos para superfícies de implantes. Objetivo: Avaliar resposta óssea na interface osso/implante, em modelo padronizado em tíbia de rato, de superfícies de implantes de titânio comercialmente puro (Ti-cp) tratadas com raloxifeno (RLX). Materiais e Métodos: Foram utilizados 144 implantes de Ti-cp, divididos em dois grupos (n=36) de acordo com o tratamento de superfície: 1-Controle (CRT); e 2-RLX. Após tratamentos, cada animal recebeu um implante em cada tíbia. Os animais foram eutanaziados após 7, 15, 30 e 40 dias para análises histológica, histométrica e imunoistoquimica. Dados quantitativos foram submetidos à ANOVA dois fatores e pós-teste de Tukey (α=0,05). Resultados: A análise histológica aos 7 dias do grupo RLX apresentou maior quantidade de tecido conjuntivo comparada ao grupo CRT. Nos demais períodos, a modelação óssea foi semelhante. A análise histométrica, em relação à área óssea neoformada (AON), apresentou diferença significante entre RLX e CRT após 7 (p=0,005) e 40 (p=0,04) dias. Em relação à análise de extensão linear de contato entre tecido ósseo e superfície do implante (ELCOI), houve diferença significante entre RLX e CRT após 7 e 15 dias (p=0,003). Na análise imunoistoquímica aos 40 dias, RLX apresentou menos matriz óssea mineralizada e AON em relação ao CRT. Conclusão: Uso do RLX para modificação de superfície de implante demonstrou características de AON semelhantes ao CRT(AU)

Background: To improve osseointegration, researches have sought implant surface treatments. Purpose: To evaluate bone response at bone/implant interface, in standardized rat tibia model, of commercially pure titanium (cp-Ti) implant treated with raloxifene (RLX). Materials and Method: 144 cp-Ti implants were divided into two groups (n=36) according to the surface treatment: 1-Control (CRT); and 2-RLX. After treatments, each animal received one implant in each tibia. The animals were euthanized after 7, 15, 30 and 40 days for histological, histometric and immunohistochemical analysis. Quantitative data were submitted to two-way ANOVA and Tukey's test (α=0.05). Results: Histological analysis at 7 days in RLX showed greater amount of connective tissue compared to CRT. In other timepoints, bone remodeling was similar. Histometric analysis, regarding the newly formed bone area (NBA), showed significant difference between RLX and CRT after 7 (p=0.005) and 40 (p=0.04) days. Regarding the analysis of linear extension of contact between bone tissue and implant surface (LECBI), there was significant difference between RLX and CRT after 7 and 15 days (p=0.003). In the immunohistochemical analysis at 40 days, RLX showed less mineralized bone matrix and NBA compared to CRT. Conclusion: Use of RLX to modify implant surface demonstrated NBA characteristics similar to CRT(AU)

Effects of raloxifene combined with low-dose conjugated estrogen on the endometrium in menopausal women at high risk for breast cancer

OBJECTIVES: To compare the effects of low-dose conjugated estrogen (CE), raloxifene, and the combination thereof on the endometrium of postmenopausal women. METHODS: Postmenopausal women between 45 and 60 years of age, with Gail score≥1.67 and no endometrial disorders, were randomly assigned to receive low-dose CE (0.3 mg), raloxifene (60 mg), or combined therapy for 1 year. Transvaginal ultrasound was performed at baseline and every 3 months; the Kupperman Index was assessed at baseline and every 6 months. Endometrial biopsies were performed if endometrial thickness (ET) was ≥5 mm or if vaginal bleeding occurred. The primary outcome was the occurrence of ET≥5 mm over the one-year period. RESULTS: Seventy-three women were randomly assigned and analyzed on an intent-to-treat basis. Eight, three, and four women in the CE, raloxifene, and combination groups, respectively, exhibited ET≥5 mm. No genital bleeding was reported in the combination group. Endometrial biopsy revealed atrophy or polyps in all groups, with one patient in the CE group exhibiting a proliferative endometrium without atypia. At 6 months, there was a progressive increase in mean ET in the CE group, but not in the other two groups, with statistically significant differences at 6, 9, and 12 months. Mean scores for vasomotor symptoms and Kupperman Index favored the CE and combination groups over raloxifene. CONCLUSION: Combined raloxifene and low-dose CE decreased the severity of menopausal symptoms to a similar extent as CE alone and had similar effects as raloxifene alone on the endometrium.

Efeito do pré-tratamento do treinamento de força e raloxifeno na periestropausa sobre a cicatrização óssea / Effect of pre-treatment of strength training and raloxifene in periestropause on bone healing

Há evidências de que o treinamento de força (TF) e raloxifeno (Ral) durante a periestropausa promove melhor qualidade óssea. No entanto, nos perguntamos se os benefícios esqueléticos do TF ou Ral realizados durante a periestropausa persistiriam após a fratura. Portanto, o presente estudo teve como objetivo analisar a influência do pré-tratamento de TF e administração de Ral durante o período de periestropausa na cicatrização óssea após osteotomia unilateral total. Durante 120 dias, ratas Wistar senescentes (18 a 21 meses completos) realizaram TF em escada três vezes por semana, receberam Ral (2,3mg/Kg/dia) via gavagem, ou realizaram TF associado ao tratamento com Ral. Após este período, os tratamentos foram interrompidos e a osteotomia total foi realizada na tíbia esquerda em todos os animais, que foram eutanasiados 1 ou 8 semanas após a cirurgia. A administração de Ral durante a periestropausa piorou o perfil bioquímico e oxidativo, diminuiu a expressão gênica de marcadores relacionados à reabsorção e remodelação óssea, que afetaram negativamente as propriedades físicoquímicas, levando a alterações na microarquitetura e massa dos calos ósseos, bem como na diminuição da resistência do calo à deformação torcional, resultando em menor qualidade do tecido durante a cicatrização óssea. Por outro lado, o TF realizado antes da osteotomia resultou em melhor cicatrização óssea, melhorando o perfil bioquímico e oxidativo, alterando o perfil genético em favor da formação e reabsorção óssea, bem como as propriedades físico-químicas do calo. Essas alterações levaram a uma melhor microarquitetura e massa óssea e aumentaram a resistência do calo à deformação torcional, confirmando seu efeito benéfico na qualidade do tecido ósseo, proporcionando aceleração da consolidação óssea.A combinação de terapias, nessa intensidade de exercício e dosagem da droga, o efeito negativo de Ral, superou o efeito positivo do TF, levando à diminuição da qualidade do tecido no processo de cicatrização óssea. Portanto, os resultados indicam que, além de excelente terapia não farmacológica e ação na prevenção da osteoporose, o TF realizado no período de envelhecimento pode aumentar a qualidade óssea no início da cicatrização e proporcionar melhor consolidação óssea. Além disso, o efeito anti-reabsortivo de Ral mostrado neste modelo atrasou o processo de reparo ósseo, trazendo considerável preocupação clínica(AU)

There is evidence that strength training (ST) and raloxifene (Ral) treatment during periestropause promotes better bone quality. We wanted to determine whether the skeletal benefits of ST or Ral treatment, performed during periestropause, would persist after fracture. Therefore, the present study aimed to analyze the influence of pretreatment with ST and administration of Ral during periestropause on bone healing after total unilateral osteotomy. Senescent female Wistar rats between 18­21 months of age, performed ST on a ladder three times per week, were administered Ral by gavage (2.3 mg/kg/day), or an association of both. After 120 days, the treatments were interrupted, and a total osteotomy was performed on the left tibia in all animals. They were euthanized 1 and 8 weeks post-osteotomy. The administration of Ral during periestropause worsened the biochemical and oxidative profile, decreased gene expression of markers related to bone resorption and remodeling, which negatively affected the physicochemical properties; this lead to changes in the bone callus microarchitecture and mass, as well as a decrease in callus resistance to torsional deformation, resulting in lower tissue quality during bone healing. In contrast, ST performed prior to the osteotomy resulted in better bone healing, improvement of the biochemical and oxidative profile, alteration of the genetic profile in favor of bone formation and resorption, as well as the physicochemical properties of the callus. These changes led to better microarchitecture and bone mass and increased callus resistance to torsional deformation, confirming its beneficial effect on the quality of bone tissue, providing acceleration of bone consolidation. The combination of therapies at this exercise intensity and drug dosage showed a negative interaction, where the negative effect of Ral overcame the positive effect of ST, leading to decreased tissue quality in the bone healing process. This study indicates that in addition to excellent nonpharmacological therapy and action in the prevention of osteoporosis, ST performed during the aging period may increase bone quality at the onset of healing and provide improved bone consolidation. Furthermore, the anti-osteoclastogenic effect of Ral shown in this model delayed the bone repair process, resulting in considerable clinical concern(AU)

LC-UV method to assay raloxifene hydrochloride in rat plasma and its application to a pharmacokinetic study

A specific, precise, and accurate LC-UV method was developed and validated to assay raloxifene hydrochloride in rat plasma. Raloxifene was analyzed after liquid-liquid extraction and quantified by reversed phase liquid chromatography (C18 column) using acetonitrile and ammonium acetate buffer 0.05 M (pH 4.0) as mobile phase at a flow rate of 1 mL.min-1 and UV detection at 287 nm. Retention times of raloxifene and internal standard (dexamethasone) were approximately 11 min and 14 min, respectively. Linearity was checked for a concentration range between 25 ng.mL-1 and 1000 ng.mL-1. Intra- and inter-day precision had relative standard deviation lower than 10% and 15%, respectively. Recovery from plasma was higher than 90%. Accuracy values were 98.21%, 99.70%, and 102.70% for lower, medium, and upper limits of quantification, respectively. Limit of quantification was 25 ng.mL-1. Drug stability was analyzed at room temperature using plasma kept in a freezer at -80 °C for 45 days after processing for 6 h and three freeze-thaw cycles. The advantages of the method developed include stability under different conditions and low limit of quantification. Its applicability was confirmed by the analysis of raloxifene levels in plasma samples in a designed pharmacokinetic study in rats after intravenous administration (5 mg.kg-1).

Raloxifene in Reducing the Risk of Postmenopausal Fracture amongst Osteoporotic Subjects

Background: Osteoporosis is a systemic skeletal disease characterized by low bone mass and microarchitectural deterioration of bone tissue, with a consequent decrease in bone mineral density (BMD) and increase in bone fragility and susceptibility to fracture. Hence; we planned the present study to assess the effect of raloxifene in reducing the risk of postmenopausal fracture amongst osteoporotic subjects. Methods: The present study included assessment of effect of raloxifene in reducing the risk of postmenopausal fracture amongst osteoporotic subjects. A total of 120 postmenopausal women were included in the present study. All the subjects were broadly divided into two broad groups with 60 subjects in each group; Group A: Subjects who were given placebo for two years Group B: Subjects who were given raloxifene 60 mg/d for 2 years. Assessment of Risk of Postmenopausal Fracture in all the subjects was done by evaluating the bone mineral density (BMD) at two years follow-up time. All the results were compiled and assessed by SPSS software. Results: Non- significant results were obtained while comparing the adverse effects among subjects of both the study groups. Overall incidence of new vertebral fractures among subjects of group A and group B included 6 and 4 percent respectively. Significant results were obtained while comparing the incidence of new vertebral fractures among subjects of group A and group B respectively. Conclusion: Significant reduction in the risk of fractures occur under the influence of raloxifene in postmenopausal women with osteoporosis.

Prescription design of raloxifene nanoemulsion / 中国药学杂志

OBJECTIVE:To study formulation design of raloxifene nanoemulsion. METHODS: The solubilities of raloxifene in excipients of nanoemulsion were investigated. On the basis, emulsifier and oil were selected by the emulsifying ability. The combination and optimum proportion among co-emulsifier, oil and emulsifier were determined by the pseudo-ternary phase diagram. The effect of raloxifene on nanoemulsion prescription was studied by drug loading. Finally chitosan and carboxylated chitosan were used to regulate the Zeta potential of raloxifene nanoemulsions. RESULTS: The optimum formulation ratio of LOA, IPP, RH40 and ethanol for loading 15 mg raloxifene is 0.167 g∶0.333 g∶0.3 g∶0.2 g, respectively. The Zeta potentials of the nanoemulsions can be changed by chitosan and carboxylated chitosan from -0.954 mV to 20 mV and -13 mV or so,respectively. The effect of different pH and dilution times on stability of formulations were slight. CONCLUSION: The formulations of raloxifene nanoemulsion including positive, negative and near zero Zeta potential were obtained, which have laid a foundation for absorption mechanism study of the raloxifene nanoemulsion.

Neuroprotective effects of selective estrogen receptor modulators in cerebral ischemia / 国际脑血管病杂志

Selective estrogen receptor modulators tamoxifen,raloxifene,and bazedoxifene reduce neuronal death through the mechanisms of anti-inflammation,antioxidant stress,and inhibition of glutamate excitotoxicity.They play a neuroprotective role in cerebral ischemia and may become a new neuroprotective agent for the treatment of ischemic stroke.

The Beneficial and Adverse Effects of Raloxifene in Menopausal Women: A Mini Review

OBJECTIVES: The present mini review aimed to summarize the existing knowledge regarding the beneficial and adverse effects of raloxifene in menopausal women. METHODS: This study is a review of relevant publications about the effects of raloxifene on sleep disorder, depression, venous thromboembolism, the plasma concentration of lipoprotein, breast cancer, and cognitive function among menopausal women. RESULTS: Raloxifene showed no significant effect on depression and sleep disorder. Verbal memory improved with administration of 60 mg/day of raloxifene while a mild cognitive impairment risk reduction by 33% was observed with administration of 120 mg/day of raloxifene. Raloxifene was associated with a 50% decrease in the need for prolapse surgery. The result of a meta-analysis showed a significant decline in the plasma concentration of lipoprotein in the raloxifene group compared to placebo (standardized mean difference, −0.43; 10 trials). A network meta-analysis showed that raloxifene significantly decreased the risk of breast cancer (relative risk, 0.572; 95% confidence interval, 0.327–0.881; P = 0.01). In terms of adverse effects of raloxifene, the odds ratio (OR) was observed to be 1.54 (P = 0.006), indicating 54% increase in the risk of deep vein thrombosis (DVT) while the OR for pulmonary embolism (PE) was 1.05, suggesting a 91% increase in the risk of PE alone (P = 0.03). CONCLUSIONS: Raloxifene had no significant effect on depression and sleep disorder but decreased the concentration of lipoprotein. Raloxifene administration was associated with an increased risk of DVT and PE and a decreased risk of breast cancer and pelvic organ prolapse in postmenopausal women.

Formulation design, absorption mechanism and bioavailability of nanoemulsions for enhancing oral absorption of raloxifene / 药学学报

Oral formulations of nanoemulsions (NE) were systematically designed, and then their effects on oral absorption of raloxifene (RAL), including their absorption mechanisms were investigated. RAL solubility in water and various excipients of NE and oil-water partition coefficient[P(O/W)] of RAL were examined. Next the optimal compatibility between emulsifiers and oils in NE were ascertained by emulsification ability. Proportions of each component and optimal RAL-NE were fully confirmed by a pseudo-ternary phase diagram and drug loading, respectively. RAL-NE quality was evaluated by particle size, zeta potential, morphology, entrapment efficiency and stability in simulated gastrointestinal fluid. A MDCK cell model was used to study the in vitro transport mechanism of RAL-NE. Oral bioavailability of RAL-NE was eventually performed in SD rats. RAL can be classified as BCSⅡ based on the solubility and P(O/W). The best formulation of RAL-NE was composed of linoleic acid (LOA):isopropyl palmitate (IPP):cremophor RH40 (RH40):alcohol as 1.67:3.33:3:2. Drug loading in pre-nanoemulsion was 15 mg·g-1 andentrapment efficiency of RAL in NE was (79.4 ±0.4)%. The particle size, zeta potential and drug content of RAL-NE were maintained in the simulated gastrointestinal fluid. The in vitro transport mechanism of RAL-NE in MDCK cells was mainly clathrin-mediated endocytosis. The oral bioavailability of RAL in RAL-NE relative to RAL-suspension was 171.9%. The best formulation of RAL-NE studied systematically was confirmed to significantly improve the RAL absorption by in vitro and in vivo evaluations (P < 0.05). This paper provides references for oral NE research and development.

A SERM increasing the expression of the osteoblastogenesis and mineralization-related proteins and improving quality of bone tissue in an experimental model of osteoporosis

Abstract Raloxifene is an antiresorptive drug, selective estrogen receptor modulator (SERM) used in the treatment of osteoporosis. Objective To evaluate proteins related to bone repair at the peri-implant bone in a rat model of osteoporosis treated with raloxifene. Material and Methods 72 rats were divided into three groups: SHAM (healthy animals), OVX (ovariectomized animals), and RLX (ovariectomized animals treated with raloxifene). Raloxifene was administered by gavage (1 mg/kg/day). Tibial implantation was performed 30 days after ovariectomy, and animals were euthanized at 14, 42, and 60 days postoperatively. Samples were collected and analyzed by immunohistochemical reactions, molecular analysis, and microtomographic parameters. Results RLX showed intense staining of all investigated proteins at both time points except for RUNX2. These results were similar to SHAM and opposite to OVX, showing mild staining. The PCR gene expression of OC and ALP values for RLX (P<0.05) followed by SHAM and OVX groups. For BSP data, the highest expression was observed in the RLX groups and the lowest expression was observed in the OVX groups (P<0.05). For RUNX2 data, RLX and SHAM groups showed greater values compared to OVX (P<0.05). At 60 days postoperatively, microtomography parameters, related to closed porosity, showed higher values for (Po.N), (Po.V), and (Po) in RLX and SHAM groups, whereas OVX groups showed lower results (P<0.05); (BV) values (P=0.009); regarding total porosity (Po.tot), RLX group had statistically significant lower values than OVX and SHAM groups (P=0.009). Regarding the open porosity (Po.V and Po), the SHAM group presented the highest values, followed by OVX and RLX groups (P<0.05). The Structural Model Index (SMI), RLX group showed a value closer to zero than SHAM group (P<0.05). Conclusions Raloxifene had a positive effect on the expression of osteoblastogenesis/mineralization-related proteins and on micro-CT parameters related to peri-implant bone healing.

Combination Therapy of Raloxifene and Alendronate for Treatment of Osteoporosis in Elderly Women

OBJECTIVES: To evaluate and compare the efficacy and safety of the combination of raloxifene and alendronate with those of monotherapies in elderly women with osteoporosis. METHODS: Sixty-two postmenopausal women (mean age 63.5 ± 0.5 years) attending gynecologic osteoporosis clinics with established osteoporosis were randomly allocated to one of four treatment groups and monitored for 3 years. All patients enrolled in this study, including those in the control group (n = 14), received 1.0 g elemental calcium and 400 units of vitamin D per day. The raloxifene group (n = 16) received raloxifene 60 mg (Evista®) per day; alendronate group (n = 17) received low-dose (5 mg) alendronate with calcitriol 0.5 µg (Maxmarvil®) per day; and the combination therapy group (n = 15) received both raloxifene 60 mg and low-dose (5 mg) alendronate with calcitriol 0.5 µg. Bone mineral density (BMD) was measured in the lumbar spine and hip before and after 3 years of treatment. RESULTS: In patients who received the combined therapy, BMD increased in the lumbar spine and the hip by 7.2% (P<0.001) and 4.8% (P<0.001) at 3 years. For patients in the alendronate group, the increases were 6.7% (P<0.001) and 3.1% (P<0.01) respectively, for the raloxifene group, the increases were 4.36% (P<0.001) and 1.9% (P<0.05) in the vertebrae and femora, respectively; however, the BMD of patients in the control group decreased by 1.81% (P<0.05) and 1.6% (P<0.05), respectively, after 3 years. Patients who received the combination therapy had significantly higher BMD in both the vertebrae femora (P<0.01) in comparison to that in those treated with raloxifene or alendronate individually. CONCLUSIONS: This 3-year randomized study showed the improved effects of alendronate and raloxifene combination on spine and hip BMD in elderly postmenopausal women with established osteoporosis.

Effects of ovarian induction with Raloxifene versus Clomiphene Citrate on endometrial receptivity in mouse endometrium / 实用医学杂志

Objective To investigate the effects of ovarian induction with raloxifene(RAL)versus clomi-phene citrate(CC)on the endometrial receptivity in mouse endometrium during perimplantation period. Methods 48 female Kun-ming mouse were randomly divided into four groups in equal number:RAL 240 mg group,RAL 180 mg group,CC group,natural conception group(NC),all treated with ovulation induction after drug administration.Successfully mated female mouse were killed,and uterus samples were collected for HE stain-ing and immunohistochemistry. Results HE staining showed that the endometrial morphology in the RAL 180 mg group and RAL 240 mg group and NC group were better than that of CC group.The expressions of COX-2 and LP-AR3 in the RAL 180 mg group and RAL 240 mg group were similar to NC group,without significant difference among the three groups(P > 0.05). But in the CC group,it was statistically significantly lower than other three groups(P<0.05),indicating ovarian induction with RAL did not decrease the expressions of COX-2 and LPAR3 in endometrium. Conclusion The mechanism of ovulation induction with RAL is similar to CC,but RAL has fewer adverse effects on the endometrial receptivity compared with CC.

Comparison of Raloxifene and Clomiphene in the Treatment of Polycystic Ovary Syndrome Complicated with Infertility / 中国药房

OBJECTIVE:To compare the clinical efficacy of raloxifene and clomiphene in the treatment of polycystic ovary syndrome(PCOS)complicated with infertility. METHODS:In retrospective study,clinical information of 137 PCOS patients with infertility in our hospital during Jun. 2012-Feb. 2015 were selected and divided into raloxifene group(70 cases)and clomiphene group(67 cases)according to therapy plan. Raloxifene group received Raloxifene tablet 120 mg at the fifth day of menstrual cy-cle,qd,for consecutive 5 d. Clomiphene group received Clomiphene citrate capsule 100 mg at the fifth day of menstrual cycle, qd,for consecutive 5 d. Both groups received B-ultrasound to detect the follicular diameter and endometrial development at the twelfth day of menstrual cycle. When endometrial thickness was ≥7 mm and the diameter of one follicle was ≥18 mm at least,all patients received abdominal subcutaneous injection of Triptorelin acetate injection 0.1 mg to induce ovulation;after 48 h,ovulation was confirmed by B ultrasound. The patients with insufficient luteal function should take dydrogesterone 10 mg as appropriate,q12 h,till 15 d after ovulation. The endometrial thickness,the number of follicles with diameter≥18 mm and follicular maturation time were compared between 2 groups on the day of injection. The sex hormone levels before treatment and on the day of injection,as well as ovulation,outcome indexes and the occurrence of ADR in 2 groups were compared. RESULTS:The endometrial thickness of raloxifene group was greater than that of clomiphene group;the number of follicles with diameter ≥18 mm was more than clo-miphene group;the follicular maturation time was shorter than clomiphene group,with statistical significance(P0.05). Compared to before treatment,there was no statistical significance in serum levels of P,E2,FSH or LH in 2 groups on the day of injection(P>0.05). Serum levels of LH in 2 groups were decreased significantly,and serum level of LH in raloxifene group was significantly lower than in clomiphene group,with statistical significance(P0.05). There was no statistical significance in the total incidence of ADR (5.71% vs. 8.96%)between 2 groups(P>0.05). CONCLUSIONS:Raloxifene has better ovulation induction effect than clomi-phene,and it can improve the sex hormone levels of PCOS patients with infertility,with few ADR,and similarity in long-term clinical effect with clomiphene.

Influência do treinamento de força associado ou não ao raloxifeno sobre o perfil transcricional e microestrutural ósseo de ratas Wistar naturalmente envelhecidas / Influence of strength training associated or not with raloxifene on transcriptional and microstructural bone profile of naturally aged female Wistar rats

A ocorrência de doenças crônicas e degenerativas é significativamente maior nos organismos durante o envelhecimento, dentre elas, a osteoporose, que resulta em aumento no número de fraturas. As fraturas são as consequências mais dramáticas da osteoporose, sendo que do colo do fêmur é a mais severa, com maior incidência de morbidades e mortalidade. A menor concentração plasmática de estrogênio nas mulheres menopausadas, exerce ação primordial no desenvolvimento desta doença. Desta maneira, o objetivo deste estudo foi estudar a prevenção da osteoporose em decorrência do envelhecimento reprodutivo feminino, especificamente no período de periestropausa, utilizando treinamento de força (TF), raloxifeno (Ral) ou combinação de TF e Ral. Durante 120 dias, ratas Wistar no período do envelhecimento (18 a 21 meses) realizaram TF em escada três vezes por semana, receberam Ral (1mg/Kg/dia) por gavagem, ou realizaram TF associado ao tratamento com Ral. Microarquitetura óssea cortical e trabecular, densidade mineral óssea areal (DMOa), força óssea, imunoistoquímica (OCN, TRAP e SOST) e superfície de osteoclastos do colo do fêmur foram avaliadas, além de PCR (Runx2, Sp7, Alp, Bsp, Ocn, Rank, Rankl, Opg, Trap e Ctsk) e Western Blot (p-ERα e TRAP) do tecido ósseo de todo o fêmur. Os resultados demonstram que os tratamentos modularam o ciclo de remodelamento ósseo de maneiras diferentes: TF estimulou RNAm de marcadores osteoblásticos e osteoclásticos, enquanto Ral diminuiu marcadores osteoclásticos e TF associado a Ral aumentou marcadores osteoblásticos e diminuiu osteoclásticos. Ambos tratamentos resultaram em melhora da microarquitetura trabecular do colo do fêmur de ratas na periestropausa, todavia, apenas o TF foi capaz de melhorar além da microarquitetura trabecular, a cortical e força óssea. Desta maneira, sugerimos que a realização de TF, utilização de Ral ou a associação de TF e Ral durante a periestropausa são intervenções válidas na prevenção de osteoporose em decorrência do envelhecimento reprodutivo feminino, porém os efeitos do TF parecem ser superiores. Levando em consideração que a carga mecânica gerada pelo TF age também em tecidos não esqueléticos, concluímos que TF pode ser intervenção sistêmica para osteoporose. Esses resultados adicionam novas informações à literatura sobre terapêuticas preventivas para osteoporose e fornecem informações relevantes para estudos pré-clínicos(AU)

The association of aging with osteoporosis results in an increased number of fractures. In these fractures, the femoral neck is involved in 75% of affected women and is one of the most dramatic possible consequences. The aim of this study was to prevent female osteoporosis using strength training (ST), raloxifene (Ral) or a combination of ST plus Ral during the natural female aging process, specifically in the periestropause period. For 120 total days, aging female Wistar rats at 18-21 months of age performed ST on three times per week, and Ral was administered daily by gavage (1mg/kg/day). Bone microarchitecture, areal bone mineral density (aBMD), bone strength of the femoral neck, immunohistochemistry, western blotting (p-ERα and TRAP) and RT-PCR were assessed. We found that the treatments modulate the bone remodeling cycle in different ways. Both ST and Ral treatment resulted in improved bone microarchitecture in the femoral neck of rats in late periestropause. However, only ST improved cortical microarchitecture and bone strength in the femoral neck. In addition, ST stimulated mRNA levels of osteoclastic and osteoblastic markers, while Ral decreased mRNA levels of osteoclastic markers. The combined ST plus Ral therapy increased osteoblastic markers and decreased osteoclast markers. In this way, we suggest that SF, the use of Ral or the association of ST and Ral during periestropause are valid interventions in the prevention of osteoporosis due to female reproductive aging, but the effects of ST seem to be superior, taking into account that the mechanical load generated by ST also acts on nonskeletal tissues, we conclude that ST can be a systemic intervention for osteoporosis. These results add new information to the literature on preventive therapies for osteoporosis and provide relevant information for preclinical studies(AU)

Ginecomastia: manifestaciones, etiología, abordaje diagnóstico y tratamiento / Gynecomastia: manifestations, etiology, diagnosis and treatment approach

La ginecomastia es una entidad de consulta frecuente, definida por la proliferación del tejido glandular mamario en los hombres, en la mayoría de los casos benigna y se puede presentar a cualquier edad. Debido a su etología variada y repercusiones tanto físicas como emocionales, debe realizarse un abordaje completo para descartar causas patológicas y realizar un tratamiento dirigido en los casos que lo ameritan.

Gynecomastia is a common condition defined as the proliferation of male breast glandular tissue. It is benign in most cases and it can appear at any age. Due to its distinct etiology and physical/psychologic outcomes, a complete approach must be performed to rule out pathologic causes, and to offer precise treatment.

Raloxifene Administration in Women Treated with Long Term Gonadotropin-releasing Hormone Agonist for Severe Endometriosis: Effects on Bone Mineral Density

OBJECTIVES: To evaluate the efficacy of raloxifene in preventing bone loss associated with long term gonadotropin-releasing hormone agonist (GnRH-a) administration. METHODS: Twenty-two premenopausal women with severe endometriosis were treated with leuprolide acetate depot at a dosage of 3.75 mg/4 weeks, for 48 weeks. Bone mineral density (BMD) was evaluated at admission, and after 12 treatment cycles. RESULTS: At cycle 12 of GnRH-a plus raloxifene treatment, lumbar spine, trochanter femoral neck, and Ward's BMD differed from before the treatment. A year after treatment, the lumbar spine and trochanter decreased slightly, but were not significantly different. CONCLUSIONS: Our study shows that the administration of GnRH-a plus raloxifene in pre-menopausal women with severe endometriosis, is an effective long-term treatment to prevent bone loss.