Analysis of Q-markers of Arisaema Cum Bile Acting on Stroke Based on UPLC-QTOF-MS/MS and TCMIP v2.0 / 中国实验方剂学杂志

ObjectiveTo predict the possible quality markers （Q-markers） of Arisaema Cum Bile in the prevention and treatment of stroke based on ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spetrometry （UPLC-QTOF-MS/MS） and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP） v2.0. MethodUPLC-QTOF-MS/MS was employed with the mobile phase of 0.1% formic acid aqueous solution （A）-0.1% formic acid acetonitrile solution （B） for gradient elution （0-3 min， 0.2%-5%B； 3-5 min， 5%-8%B； 5-8 min， 8%-10%B； 8-14 min， 10%-25%B； 14-18 min， 25%-50%B； 18-20 min， 50%-70%B； 20-21 min， 70%-98%B； 21-23 min， 98%B； 23-24 min， 98%-0.2%B； 24-26 min， 0.2%B）， the flow rate of 0.5 mL·min-1 and electrospray ionization （ESI）. High quality MS/MS data were scanned in positive and negative ion modes with scanning range of m/z 50-1 500. A local database of the chemical constituents in Arisaema Cum Bile was established by UNIFI 1.8. Then the chemical constituents in Arisaema Cum Bile were characterized by matching with the local database and comparing with the reference substances and literature information. TCMIP v2.0 was used to obtain the targets corresponding to the identified components of Arisaema Cum Bile and stroke， and the "disease-formula" correlation analysis was carried out to screen the core targets by topological eigenvalues. DAVID 6.8 was used for enrichment analysis of Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway of core targets. According to the "five principles" of Q-markers and combined with literature reports， the Q-markers of Arisaema Cum Bile in the prevention and treatment of stroke were predicted， and the core components acting on these target genes were obtained. Cytoscape 3.8.0 was employed to draw the network diagram of "medicinal materials-active ingredients-target genes-pathways". Finally， AutoDock Vina 1.2.2 was used to calculate and verify the molecular docking between the candidate components and the key targets. ResultA total of 76 chemical components was identified in positive and negative ion modes， 85 core targets were collected for Arisaema Cum Bile in the prevention and treatment of stroke. A total of 31 stroke-related pathways， 23 target genes and 9 main active components of Arisaema Cum Bile acting on these genes were screened， and then we determined 4 possible Q-markers for Arisaema Cum Bile in the prevention and treatment of stroke according to the "five principles". ConclusionThe possible Q-markers of Arisaema Cum Bile for stroke are gallic acid， apigenin-6，8-di-C-glucoside， apigenin and cholic acid， and the target of these four components may be estrogen receptor alpha （ESR1）.

Estrogen Receptor-A in Medial Preoptic Area Contributes to Sex Difference of Mice in Response to Sevoflurane Anesthesia / 神经科学通报·英文版

A growing number of studies have identified sex differences in response to general anesthesia; however, the underlying neural mechanisms are unclear. The medial preoptic area (MPA), an important sexually dimorphic structure and a critical hub for regulating consciousness transition, is enriched with estrogen receptor alpha (ERα), particularly in neuronal clusters that participate in regulating sleep. We found that male mice were more sensitive to sevoflurane. Pharmacological inhibition of ERα in the MPA abolished the sex differences in sevoflurane anesthesia, in particular by extending the induction time and facilitating emergence in males but not in females. Suppression of ERα in vitro inhibited GABAergic and glutamatergic neurons of the MPA in males but not in females. Furthermore, ERα knockdown in GABAergic neurons of the male MPA was sufficient to eliminate sex differences during sevoflurane anesthesia. Collectively, MPA ERα positively regulates the activity of MPA GABAergic neurons in males but not in females, which contributes to the sex difference of mice in sevoflurane anesthesia.

Câncer gástrico com expressão positiva do receptor de estrogênio alfa: uma série de casos de um único centro ocidental / Gastric cancer with positive expression of estrogen receptor alpha: a case series from a single western center

RESUMO - RACIONAL: Apesar do avanço nas terapias, o prognóstico de pacientes com câncer gástrico (CG) avançado permanece ruim. Vários estudos demonstraram a expressão do receptor de estrogênio alfa (REa), porém seu significado no CG permanece controverso. OBJETIVO: relatar uma série de casos de CG com expressão de REa-positivo, e descrever suas características clínicopatológicas e prognóstico. MÉTODOS: Avaliamos retrospectivamente os pacientes com CG submetidos à gastrectomia com intenção curativa entre 2009 e 2019. A expressão do REa foi avaliada por imuno-histoquímica por meio da construção de microarranjos de tecido (TMA). Pacientes com adenocarcinoma gástrico ERa-negativos serviram como grupo comparação. RESULTADOS: No período selecionado, foram identificados 6 (1,8%) CG REa-positivos entre os 345 CG analisados. Todos os ERa-positivos eram homens, com idades entre 34-78 anos, tinham CG do tipo difuso de Lauren e pN+. Comparado aos REa-negativos, os CG REa-positivos associaram-se a maior diâmetro (p=0,031), gastrectomia total (p=0,012), tipo de Lauren difuso/misto (p=0,012), presença de invasão perineural (p=0,030) e metástase linfonodal (p=0,215). O estágio final foi o IIA em um caso; IIIA em três e IIIB em dois casos. Entre os 6 pacientes REa -positivos, 3 tiveram recorrência da doença (peritoneal) e morreram. Não houve diferença significativa na sobrevida entre os grupos REa-positivo e negativo. CONCLUSÃO: A expressão do REa é menos comum no CG, estando associada à histologia difusa e presença de metástases linfonodal, podendo servir como um marcador relacionado à progressão tumoral e pior prognóstico. Além disso, uma alta taxa de recorrência peritoneal foi observada em pacientes ERa-positivos.

ABSTRACT - BACKGROUND: Despite advances in therapies, the prognosis of patients with advanced gastric cancer (GC) remains poor. Several studies have demonstrated the expression of estrogen receptor alpha (ERa); however, its significance in GC remains controversial. AIM: The present study aims to report a case series of GC with ERa-positive expression and describe their clinicopathological characteristics and prognosis. METHODS: We retrospectively evaluated patients with GC who underwent gastrectomy with curative intent between 2009 and 2019. ERa expression was assessed by immunohistochemistry through tissue microarray construction. Patients with ERa-negative gastric adenocarcinoma served as a comparison group. RESULTS: During the selected period, 6 (1.8%) ERa-positive GC were identified among the 345 GC patients analyzed. All ERa-positive patients were men, aged 34-78 years, and had Lauren diffuse GC and pN+ status. Compared with ERa-negative patients, ERa-positive patients had larger tumor size (p=0.031), total gastrectomy (p=0.012), diffuse/mixed Lauren type (p=0.012), presence of perineural invasion (p=0.030), and lymph node metastasis (p=0.215). The final stage was IIA in one case, IIIA in three cases, and IIIB in two cases. Among the six ERa-positive patients, three had disease recurrence (peritoneal) and died. There was no significant difference in survival between ERa-positive and ERa-negative groups. CONCLUSIONS: ERa expression is less common in GC, is associated with diffuse histology and presence of lymph node metastasis, and may be a marker related to tumor progression and worse prognosis. Also, a high rate of peritoneal recurrence was observed in ERa-positive patients.

Expression of estrogen receptor alpha protein and BRAF V600E protein in thyroid papillary carcinoma and its clinical significance / 中华普通外科杂志

Objective:To investigate the expression of estrogen receptor alpha (ERα) protein and BRAF V600E protein in thyroid papillary carcinoma (PTC) and their relationship with clinical factors of PTC. Methods:The expression of ERα and BRAF V600E protein in 1 105 PTC patients was detected by immunohistochemistry. The relationship among ERα, BRAF V600E protein and clinical factors were analyzed. Results:Positive ERα protein was correlated with maleness (χ 2= 6.087, P=0.001), age< 45 years old (χ 2=5.197, P=0.023) and multifocal tumors (χ 2=4.446, P=0.035). Positive BRAF V600E protein was correlated with positive ERα protein (χ 2=6.209, P=0.013), Hashimoto thyroiditis (χ 2=29.388, P<0.001), no lateral lymph node metastasis (χ 2=6.849, P=0.009) and multifocal tumors (χ 2=9.596, P=0.035). Conclusions:ERα expression is more common in male patients, patients younger than 45 years of age, those with multifocal tumors and positive BRAF V600E protein. BRAF V600E protein may inhibit Hashimoto's thyroiditis, tumor growth and the occurrence of lateral lymph node metastasis, and promote the occurrence of multiple focal tumors.

Role of ERRα in regulating ATP synthesis in hiPSCs-derived cardiomyocytes / 解放军医学杂志

Objective: To investigate the role of estrogen-related receptor alpha (ERRα) in regulating the adenosine triphosphate (ATP) synthesis in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Methods: Undifferentiated human induced pluripotent stem cells (hiPSCs) were induced to differentiate into cardiomyocytes by sequential transient activation/inhibition of Wnt signaling pathway. Immunofluorescence was used to detect the expression of pluripotency markers sex determining region Y-box 2 (SOX2), stage specific embryonic antigen 4 (SSEA4) and tumor resistance antigen 1-60 (TRA-1-60) in hiPSCs and the expression of cardiac specific markers cardiac troponin T (cTnT) and connexin 43 (Cx43) in hiPSCCMs, respectively; RT-qPCR was used to detect the mRNA expression levels of cardiac troponin T2 (TNNT2) and myosin heavy chain 6 (MYH6) in hiPSCs and hiPSCs-CMs; Western blotting was performed to detect the protein expression levels of ERRα, cytochrome C (CytC) and mitochondrial pyruvate carrier 1 (MPC1) in hiPSCs-CMs. Additionally, the ERRα-specific inhibitor XCT790 was used to treat the hiPSC-CMs, and then the protein expressions of ERRα, CytC and MPC1 were detected by Western blotting, and the changes of cell viability, intracellular ATP content and mitochondrial membrane potential were measured by assay kits. Results: Immunofluorescence results showed that hiPSCs expressed SOX2, SSEA4 and TRA-1-60, while hiPSC-CMs expressed cTnT and Cx43; compared with hiPSCs, the mRNA levels of TNNT2 and MYH6 in hiPSC-CMs increased significantly (82.820 ± 2.005 vs. 1.001 ± 0.029, 90982.000 ± 1968.000 vs. 1.003 ± 0.053, respectively, P<0.05), and intracellular ATP content and protein expression levels of ERRα, CytC and MPC1 also increased significantly [(9.905 ± 1.286) nmol/mg protein vs. (4.582 ± 0.141) nmol/mg protein, 5.392 ± 0.313 vs. 1.050 ± 0.076, 8.954 ± 0.293 vs. 1.071 ± 0.067, 2.605 ± 0.088 vs. 1.031 ± 0.091, respectively] with significant differences (P<0.05). Furthermore, compared with the control group, 10 μmol/L XCT790 could effectively inhibit the protein activity of ERRα in hiPSC-CMs without cytotoxicity, and reduced intracellular ATP content and mitochondrial membrane potential [(4.903 ± 1.158) nmol/mg protein vs. (9.310 ± 0.980) nmol/mg protein, 1.407 ± 0.022 vs. 1.977 ± 0.093, respectively], meanwhile down-regulated the protein expression levels of MPC1 and CytC in hiPSC-CMs (0.705 ± 0.019 vs. 0.897 ± 0.011, 0.594 ± 0.021 vs. 0.797 ± 0.025, respectively, P<0.05). Conclusions: The increase of ATP content after differentiation of hiPSCs into cardiomyocytes is related to the increase of ERRα expression. In hiPSC-CMs, ERRα may regulate the ATP synthesis though regulating the mitochondrial membrane potential and the protein expression of CytC and MPC1.

Mechanism of traditional Chinese medicine balancing Yin-Yang by targeting ERα/ERβ and its application in treatment of menopausal syndrome / 中国中药杂志

The coordination and unification of Yin and Yang are the basis of normal human life activities. Along with the age growth and aging of the body, women will suffer from menopausal syndrome during menopause. In addition to the significant changes in the genital system, there are also pathological manifestations in estrogen target points including bone, nerve and cardiovascular systems, due to the imbalance of Yin and Yang. Besides the insufficiency of estrogen, the main cause of menopausal syndrome is the changes in the response of target organs to estrogen. In other words, the biological effects mediated by estrogen receptor(ER) alpha and beta subtypes in target cells are often different or even opposite; the changes of expression level and ratio of ERα and ERβ are also important causes for the abnormal estrogenic effects in target organs and the imbalance of Yin and Yang of the body. Therefore, on one hand, the therapeutic mechanism of drugs is ER-mediated estrogenic effect. On the other hand, the drugs have a regulatory effect on ER subtype expression in target cells and Yin-Yang state in target organs and even organisms, so as to cause further changes in the response of target cells to estrogen or estrogenic components, and exert its therapeutic effects. This paper reviews the pharmacological mechanism of gynecological traditional Chinese medicine in harmonizing Yin and Yang in estrogen-positive target cells and the clinical efficacy in the following aspects, including estrogen and its mechanism, the estrogenic effect of ER in traditional Chinese medicine and the mechanism of ER subtype in balancing Yin and Yang and mediating and regulating the main target tissues in menopausal syndrome treatment.

Association of estrogen receptor α gene polymorphisms with hepatitis B cirrhosis / 临床肝胆病杂志

At present, hepatitis B cirrhosis still has a major impact on public health and national development. The development and progression of hepatitis B cirrhosis is associated with estrogen. Estrogen acts by binding to estrogen receptors, and the expression and functional status of estrogen receptors are affected by estrogen receptor gene polymorphisms. This article mainly introduces the structure and biological characteristics of estrogen and estrogen receptor α gene and their association with the pathogenesis of hepatitis B cirrhosis, and it is pointed out that the abnormal expression of estrogen may be associated with the development and progression of hepatitis B cirrhosis. However, due to the differences in genetic loci, environment, and location between studies, the results of the association of estrogen receptor α gene polymorphisms with hepatitis B cirrhosis may not be completely consistent.

Current status and future of non-genomic effects of phytoestrogens / 药学学报

Phytoestrogens exhibit various pharmacological estrogen-like effects, such as in the prevention and treatment of osteoporosis, cardiovascular diseases, tumors, etc., but the specific mechanism is still unclear. In recent years, estrogen receptor alpha-mediated rapid non-genomic effects have been identified to play an important role in the pathogenesis of estrogen-related diseases. The research of phytoestrogens exerting pharmacological effects through non-genomic effects has also received increasing attention. This article summarizes the research progress in estrogen receptor alpha-mediated non-genomic effects and analyzes the possible involvement of rapid non-genomic effects in certain pharmacological effects of phytoestrogens. The future prospects of estrogen receptor-mediated non-genomic effects by phytoestrogens are also discussed.

Effect of triptolide on immune function and tumor cell proliferation in patients cervical cancer / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To explore the effects of triptolide on immune function and tumor cell proliferation in patients with cervical cancer. Methods: Sixty-two patients with cervical cancer admitted in the Department of Obstetrics and Gynecology of the Second Affiliated Hospital of Hebei North College between July 2015 and April 2018 were randomly divided into the control group (n=31) and the observation group (n=31). All patients received routine treatment after laparoscopy, while those in the observation group received additional triptolide. The treatment efficacy, serum immune cells, inflammatory factors and the levels of cyclinD1, estrogen receptor α (ERα ) were observed and compared between the two groups. Results: The total remission rate of the patients in the observation group was 87.10%, significantly higher than 61.29% in the control group (P<0.05). After treatment, the levels of CD3+ and CD4+ T lymphocytes and CD4+ /CD8+ T lymphocytes in the two groups increased significantly, with more obvious increase in the observation group than that in the control group (P<0.01). The levels of CD8+ and programmed cell death-ligand 1 (PD-L1) T lymphocytes in the two groups decreased significantly after treatment, with a more obvious decrease in observation group than that in control group (P<0.01). After treatment, the levels of interleukin-6 (IL-6), IL-10 and tumor necrosis factor-α (TNF-α) in the two groups decreased, and those in the observation group were significantly lower than those in the control group (P<0.05). After treatment, the positive expression rate of cyclinD1 decreased and the positive expression rate of ER α increased in both groups (all P<0.05), with no significant difference between the two groups (all P>0.05). Conclusion: On the basis of routine surgical treatment, triptolide can effectively improve the immune function, reduce the inflammatory response, inhibit the proliferation of tumor cells and regulate the expression of cancer-related factors in patients with cervical cancer, which has a certain therapeutic effect on cervical cancer.

Association between decreased expression of estrogen receptor alpha, androgen receptor and phosphatase and tensin homolog immunoexpression in the canine prostate / Associação entre diminuição da expressão dos receptores de estrógeno alfa e andrógeno, e imunoexpressão de fosfatase e tensina homóloga na próstata canina

Canine prostate gland is a hormonal dependent organ and its imbalance of estrogen and androgen receptor expressions are directly associated with the development of different diseases. Due to the lack of information regarding the behavior of the aforementioned receptors in canine prostate cancer (PC), this study aimed to identify estrogen receptor alpha (ERα), androgen receptor (AR), Ki67 and phosphatase and tensin homolog (PTEN) protein expressions in canine PC by immunohistochemistry. We found nuclear expression of ERα and AR in the epithelial cells of normal canine samples and a loss of protein expression in PC samples. Normal samples showed Ki67 expression in a few basal cells and the PC samples showed the highest mean of positive cells (253.1). Canine prostate cancer showed a high proliferative index, which was associated with independence of hormonal actuation. PTEN showed positive nuclear and cytoplasmic expression in normal canine samples and a loss in PC. Loss of ERα, AR and PTEN indicated that canine PC exhibits the same immunohistochemical phenotype as in human patients with PC resistant to hormonal therapy. Therefore, canine PC should be considered as a model to study human PC resistant to hormonal therapy.(AU)

A glândula prostática canina é um órgão dependente de hormônio, e o desequilíbrio na expressão dos receptores de estrógeno e andrógeno estão diretamente associados com o desenvolvimento de diferentes doenças. Devido à falta de informação sobre o comportamento desses receptores no câncer prostático canino (PC), este estudo tem por objetivo identificar a expressão proteica através da técnica de imuno-histoquímica do receptor de estrógeno alfa (REα), receptor de andrógeno (RA), Ki67 e fosfatase e tensina homóloga (PTEN). Foi encontrado nas células epiteliais prostáticas normais caninas a expressão nuclear de REα e RA, e perda de expressão proteica nas amostras de PC. As amostras normais apresentaram expressão de Ki67 em poucas células basais e as amostras de PC apresentaram a maior média de células positivas (253,1). O câncer de próstata canino apresentou uma taxa alta de proliferação, o qual foi associado com a atuação independente de hormônio. As amostras de próstatas caninas normais revelaram marcação nuclear e citoplasmática da proteína PTEN e perda nas amostras de PC. A perda de REα, RA e PTEN indicam que as amostras de PC exibem o mesmo fenótipo imuno-histoquímico de pacientes humanos com câncer prostático resistente a terapia hormonal. Sendo assim, o PC canino deve ser considerado um modelo para estudos de câncer prostático humano resistente a terapia hormonal.(AU)

The correlation between Erα gene polymorphism and the distribution pattern of fascia syndrome in women with knee osteoarthritis / 国际中医中药杂志

Objective@#To study the correlation between Erα gene polymorphism and the distribution pattern of TCM meridian syndrome in women with knee osteoarthritis.@*Methods@#A total of 120 Knee osteoarthritis (KOA) female patients admitted to our hospital between January 2018 and December 2018 were selected as the observation group, and 120 healthy female subjects accepted physical examination at our hospital over the same period were selected as the control group. The Erα gene distribution of estrogen receptor was compared between the two groups to identify the differences between KOA women and healthy women. The TCM syndromes and meridians types of the patients in the observation group were defined to evaluate the degree of KOA. The correlation among KOA status, Erαgenotypes, TCM syndromes and meridians was analyzed.@*Results@#The distribution of Erα gene XbaI was significantly different between the observation group and the control group (χ2=35.339, P<0.001). There was a statistical difference in the distribution of XbaI genotypes between mild to moderate and severe patients in the observation group (χ2=29.088, P<0.001). There was a moderate correlation between the Lequesne score and the fascia type in KOA women, and a high correlation between the Lequesne score and the TCM syndrome differentiation type, and a moderate correlation between the Lequesne score and the Erα gene and the XbaI genotype (P<0.05). The Erα gene polymorphisms, meridians and syndrome differentiation were moderately correlated (P<0.05).@*Conclusions@#In female patients with knee osteoarthritis, the conditions are more serious in patients with complex type of tendons, deficiency of liver and kidney, phlegm and blood stasis, Erα gene xx genotype, and there is a clear correlation between the TCM type of tendons and ER gene polymorphism.

Effects of estrogen on bicarbonate secretion of duodenal mucosa / 中华消化杂志

Objective@#To study the effects of estrogen on bicarbonate secretion of duodenal mucosal, and to observe estrogen receptor (ER) subtypes of estrogen.@*Methods@#Sixteen 4-week-old male C57 mice were divided into control group and estrogen group, with eight mice in each group. The mice serum level of estrogen was detected by chemiluminescence. The expression of cystic fibrosis transmembrane conductance regulator (CFTR), solute carrier family 26 (SLC26) A3 and SLC26A6 in the duodenum tissues were determined by real-time polymerase chain reaction (RT-PCR). After SCBN cells treated with estrogen, ERα and ERβ blocking agent, and transfected with silenced ERα and ERβ for 24 and 48 hours, the expression levels of CFTR, SLC26A3 and SLC26A6 mRNA in cells were detected by RT-PCR. The effects of estrogen before and after silenced ERα and ERβ on bicarbonate secretion of SCBN cells were observed by high-speed ion imaging system. T test and rank sum test were used for statistical analysis.@*Results@#Compared with that of control group, the serum estrogen level of estrogen group was significantly high ((4 874±942) pmol/L vs. (657±187) pmol/L, t=-11.579, P<0.01). The expression levels of CFTR, SLC26A3 and SLC26A6 mRNAs in duodenum tissues of estrogen group were higher than those of control group (0.856±0.302 vs. 0.452±0.246, 2.910±1.680 vs. 1.120±0.540, 1.272±0.667 vs. 0.319±0.114), and the differences were statistically significant (t=-2.317, -2.483 and -3.721, all P<0.05). Compared with those treated with estrogen for 24 and 48 hours, the levels of CFTR mRNA and SLC26A6 mRNA were lower after the ERβ blocking agent were added into estrogen for 24 and 48 hours (CFTR mRNA: 0.171±0.059 vs. 0.522±0.260 and 0.111±0.014 vs. 0.578±0.297; SLC26A6 mRNA: 0.486±0.289 vs. 1.118±0.571 and 0.492±0.231 vs. 1.551±0.605), and the differences were statistically significant (tCFTR mRNA=2.974 and 2.655, tSLC26A6 mRNA=2.393 and 3.272; all P<0.05). Compared with those of silenced ERα group, the levels of CFTR mRNA, SLC26A3 mRNA and SLC26A6 mRNA were higher after ERα silenced and then added estrogen for 24 and 48 hours (24 h: 5.073±2.270 vs. 1.185±0.494, 1.796±1.168 vs. 0.468±0.108 and 3.085±1.357 vs. 0.706±0.347; 48 h: 5.025±1.998 vs. 1.185±0.494, 1.557±0.653 vs. 0.468±0.108 and 3.290±1.750 vs. 0.706±0.347), and the differences were statistically significant (t24 h=-4.122, -2.773 and -4.162, t48 h=-4.604, -4.034 and -3.250; all P<0.05). Compared with that of silenced ERα group, the bicarbonate secretion increased after ERα silenced and then added estrogen for 24 and 48 hours (0.72±0.17 and 1.15±0.25 vs. 0.35±0.17), and pH also elevated, and the differences were statistically significant (t=-6.516 and -12.387, both P<0.01).@*Conclusion@#Estrogen mainly up-regulates the expression of bicarbonate transporter protein in duodenal mucosal epithelial cells through ERβ, and promotes the bicarbonate secretion of duodenal mucosa.

Effects of estrogen on bicarbonate secretion of duodenal mucosa / 中华消化杂志

Objective To study the effects of estrogen on bicarbonate secretion of duodenal mucosal , and to observe estrogen receptor (ER) subtypes of estrogen.Methods Sixteen 4-week-old male C57 mice were divided into control group and estrogen group , with eight mice in each group .The mice serum level of estrogen was detected by chemiluminescence .The expression of cystic fibrosis transmembrane conductance regulator (CFTR), solute carrier family 26 (SLC26) A3 and SLC26A6 in the duodenum tissues were determined by real-time polymerase chain reaction ( RT-PCR).After SCBN cells treated with estrogen , ERαand ERβblocking agent, and transfected with silenced ER αand ERβfor 24 and 48 hours, the expression levels of CFTR, SLC26A3 and SLC26A6 mRNA in cells were detected by RT-PCR.The effects of estrogen before and after silenced ER αand ERβon bicarbonate secretion of SCBN cells were observed by high-speed ion imaging system.T test and rank sum test were used for statistical analysis .Results Compared with that of control group , the serum estrogen level of estrogen group was significantly high ((4 874 ±942) pmol/L vs.(657 ±187) pmol/L,t＝-11.579, P?0.01). The expression levels of CFTR, SLC26A3 and SLC26A6 mRNAs in duodenum tissues of estrogen group were higher than those of control group (0.856 ±0.302 vs.0.452 ±0.246, 2.910 ±1.680 vs.1.120 ±0.540, 1.272 ± 0.667 vs.0.319 ±0.114), and the differences were statistically significant ( t ＝-2.317,-2.483 and-3.721, all P?0.05).Compared with those treated with estrogen for 24 and 48 hours, the levels of CFTR mRNA and SLC26A6 mRNA were lower after the ERβblocking agent were added into estrogen for 24 and 48 hours (CFTR mRNA: 0.171 ±0.059 vs.0.522 ±0.260 and 0.111 ±0.014 vs.0.578 ±0.297; SLC26A6 mRNA:0.486 ±0.289 vs.1.118 ±0.571 and 0.492 ±0.231 vs.1.551 ±0.605), and the differences were statistically significant (tCFTR mRNA＝2.974 and 2.655, tSLC26A6 mRNA＝2.393 and 3.272; all P?0.05).Compared with those of silenced ERαgroup, the levels of CFTR mRNA, SLC26A3 mRNA and SLC26A6 mRNA were higher after ERα silenced and then added estrogen for 24 and 48 hours (24 h: 5.073 ±2.270 vs.1.185 ±0.494, 1.796 ±1.168 vs.0.468 ±0.108 and 3.085 ±1.357 vs.0.706 ±0.347; 48 h: 5.025 ±1.998 vs.1.185 ±0.494, 1.557 ± 0.653 vs.0.468 ±0.108 and 3.290 ±1.750 vs.0.706 ±0.347), and the differences were statistically significant (t24 h＝-4.122,-2.773 and -4.162, t48 h ＝-4.604,-4.034 and -3.250; all P?0.05).Compared with that of silenced ERαgroup, the bicarbonate secretion increased after ER αsilenced and then added estrogen for 24 and 48 hours (0.72 ±0.17 and 1.15 ±0.25 vs.0.35 ±0.17), and pH also elevated, and the differences were statistically significant (t＝-6.516 and -12.387, both P?0.01).Conclusion Estrogen mainly up-regulates the expression of bicarbonate transporter protein in duodenal mucosal epithelial cells through ER β, and promotes the bicarbonate secretion of duodenal mucosa .

Effects of metformin on the expression of estrogen synthetase and ER mRNA in uterine leiomyoma tissues / 中华妇产科杂志

Objective To elucidate whether metformin could regulate the mRNA expression level of estrogen synthetase and ER in human uterine leiomyoma tissues. Methods (1) Seventeen pairs of uterine leiomyoma tissues and adjacent myometrium (>2 cm) were collected from patients underwent hysterectomy in Peking University First Hospital between December 2016 and January 2017. Real-time PCR was used to measure the mRNA expression level of estrogen synthetase [including cytochrome P450 cholesterol side chain cleavage enzyme (P450scc), cytochrome P450 17 α-hydroxylase (P450c17), 3-beta-hydroxysteroid dehydrogenase type 2 (3β-HSD-2), 17-beta-hydroxysteroid dehydrogenase type 1 (17β-HSD-1) and aromatase cytochrome P450 (P450arom)] and ER (including ERα and ERβ) in the uterine leiomyoma tissues and adjacent myometrium. (2) Uterine leiomyoma cells derived from uterine leiomyoma tissues were identified by immunocytochemistry method and cultured to the third generation. The treatment groups were cultured with different concentrations of metformin (10, 50 and 100 μmol/L) for 48 hours, and the control group was cultured with deionized water for 48 hours. The mRNA expression level of estrogen synthetase and estrogen receptor subtypes were measured by real-time PCR. Results (1) P450scc, P450c17, 3β-HSD-2, 17β-HSD-1, P450arom mRNA median expression levels were 112, 4, 13, 42 and 194 in the uterine leiomyoma tissues, and were respectively 114, 5, 11, 32 and 6 in the myometrium. Compared to those of the myometrium, 3β-HSD-2 and P450arom mRNA expression levels in the uterine leiomyoma tissue were significantly higher (P<0.05), while there were no significant change of mRNA expression levels among P450scc, P450c17 and 17β-HSD-1 (P>0.05). ERα and ERβ mRNA median expression levels were 208 and 116 in the uterine leiomyoma tissues, and were 24 and 95 in the myometrium. Compared to that of the myometrium, ERα mRNA level in the uterine leiomyoma tissue was significantly higher (P=0.001), while there were no significant change of ERβ mRNA level (P=0.193). (2) After cultured with different concentrations of metformin (10, 50 and 100 μmol/L), the P450arom mRNA levels in the uterine leiomyoma tissues were 9 ± 4, 8 ± 5 and 8 ± 3 respectively in the treatment groups and was 16 ± 5 in the control group. Compared to that of the control group, P450arom mRNA expression levels in the treatment groups were significantly declined (P<0.05). There were no significant different change of mRNA expression levels among 3β-HSD-2, ERα and ERβ between the treatment groups and the control group (P>0.05). Conclusions Metformin could down-regulate the mRNA expression level of aromatase in the uterine leiomyoma cells. These results indicate that metformin may inhibit the local estrogen synthesis and therefore suppress the development of uterine leiomyoma.

Evaluation of estrogen receptor expression in low-grade and high-grade astrocytomas

SUMMARY OBJECTIVE: This study aims to compare estrogen receptor expression between low and high-grade astrocytomas. METHOD: A study using paraffin blocks of glial tumors from the Anatomy Pathology archives of São Marcos Hospital was carried out and began after approval by the Review Board of the Federal University of Piaui. Specimens were histochemically marked with an anti-ER alpha antibody. Brown-stained nuclei were considered positive, regardless of reaction intensity. Data were statistically analyzed using the Mann-Whitney test and Spearman's correlation. Statistical significance was established at p<0.05. RESULTS: The mean percentage of nuclei stained with anti-ER alpha in low-and high-grade astrocytomas was 0.04 and zero, respectively, while Spearman's correlation showed a strong negative association between low and high-grade tumors (p<0.001) and (r= −0.67), respectively. CONCLUSION: In the current study, estrogen receptor expression was positive only in low-grade astrocytomas and nil in high-grade astrocytomas, showing that ER expression declines with the grade of tumor malignancy.

RESUMO OBJETIVO: O objetivo deste estudo é comparar a expressão do receptor de estrogênio entre astrocitomas de baixo e alto grau. MÉTODO: Foi realizado um estudo usando blocos de parafina de tumores gliais dos arquivos de Anatomia Patológica do Hospital São Marcos e iniciado após aprovação pelo Comitê de Ética da Universidade Federal do Piauí. Os espécimes foram marcados histoquimicamente com anticorpo anti-ER alpha. Os núcleos corados em marrom foram considerados positivos, independentemente da intensidade da reação. Os dados foram analisados estatisticamente utilizando o teste de Mann-Whitney e a correlação de Spearman. A significância estatística foi estabelecida em p<0,05. RESULTADOS: A porcentagem média de núcleos corados com anti-ER alfa em astrocitomas de baixo e alto grau foi de 0,04 e zero, respectivamente, enquanto a correlação de Spearman mostrou uma forte correlação negativa entre tumores de baixa e alta qualidade (p<0,001) e (r=-0,67), respectivamente. CONCLUSÕES: No presente estudo, a expressão do receptor de estrogênio foi positiva apenas em astrocitomas de baixo grau e nula em astrocitomas de alto grau, mostrando que a expressão de ER diminui com o grau de malignidade tumoral.

Estrogen receptor-alpha gene XbaI A > G polymorphism influences short-term cognitive decline in healthy oldest-old individuals / Polimorfismo XbaI A > G no gene do receptor do estrogênio-alfa influencia o declínio cognitivo em curto prazo em idosos muito idosos saudáveis

ABSTRACT This prospective study aimed to evaluate the influence of the -351A/G XbaI polymorphism in the estrogen receptor-alpha (ESR-1) gene on global cognitive scores of a community sample of healthy oldest-old individuals within one year of follow up. Methods The individuals were categorized in two groups according to the presence or absence of cognitive decline. Cognitive data were related to genetic information. Results The XbaI -351 AA genotype was more common among cognitive decliners, while -351G allele carriers showed cognitive stability or improvement. Conclusion These results suggest that ESR-1 could be associated with one-year cognitive decline in healthy oldest-old individuals, since the estrogen pathway may be involved with neuroprotection, even in healthy brain aging.

RESUMO Neste estudo prospectivo foi avaliada a influência do polimorfismo -351A/G XbaI do gene do receptor de estrogênio alfa (ESR-1) sobre o desempenho cognitivo global em idosos muito idosos (≥ 75 anos) saudáveis durante um ano. Métodos Os indivíduos foram divididos em dois grupos de acordo com a presença ou ausência de declínio cognitivo. Dados cognitivos foram relacionados à informação genética. Resultados O genótipo XbaI -351 AA foi mais comum entre indivíduos que apresentaram declínio cognitivo, enquanto carreadores do alelo -351G demonstraram estabilidade ou melhora cognitiva. Conclusão Estes resultados sugerem que ESR-1 poderia estar associado ao declínio cognitivo em curto prazo em idosos saudáveis, possivelmente por meio de propriedades neuroprotetoras do estrogênio, mesmo em cérebros idosos saudáveis.

Laminin Modulates the Stem Cell Population in LM05-E Murine Breast Cancer Cells through the Activation of the MAPK/ERK Pathway / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: We investigated the effects of laminin on the fraction of cells with self-renewing capacity in the estrogen-dependent, tamoxifen-sensitive LM05-E breast cancer cell line. We also determined whether laminin affected the response to tamoxifen. MATERIALS AND METHODS: The LM05-E breast cancer cell line was used as a model for all experiments. Aldehyde dehydrogenase (ALDH) activity, clonogenic and mammosphere assays were performed to measure the effects of laminin on modulation of the stem cell subpopulation. Pluripotent gene expression was analyzed by reverse transcriptase–polymerase chain reaction. The involvement of the mitogen-activated protein kinase (MAPK)/ERK pathway was determined using specific inhibitors. The effects of laminin on the response to tamoxifenwere determined and the involvement of α6 integrin was investigated. RESULTS: We found that pretreatment with laminin leads to a decrease in cells with the ability to form mammospheres that was accompanied by a decrease in ALDH activity. Moreover, exposure of mammospheres to laminin reduced the capacity to form secondary mammospheres and decreased the expression of Sox-2, Nanog, and Oct-4. We previously reported that 4-OH-tamoxifen leads to an increase in the expression of these genes in LM05-E cells. Treatment with signaling pathway inhibitors revealed that the MAPK/ERK pathway mediates the effects of laminin. Finally, laminin induced tamoxifen resistance in LM05-E cells through α6 integrin. CONCLUSION: Our results suggest that the final number of cells with self-renewing capacity in estrogen-dependent breast tumors may result from the combined effects of endocrine treatment and microenvironmental cues.

Effects of acacetin on T47D cell proliferation / 中国药理学通报

Aim To investigate the effect of acacetin on cell proliferation and the influence of acacetin on estrogen receptor expression in vitro.Methods The proliferation rates and the cell cycle changes of acace-tin-treated T47D cells were measured by sulforhodam-ine B(SRB)assay and flow cytometry,respectively. Moreover,the mRNA expressions of estrogen receptor-alpha(ERα),estrogen receptor-beta(ERβ)and pro-liferating antigen(Ki67)were determined by quantita-tive real time PCR (qPCR).Western blot was em-ployed to detect the ERαand ERβprotein expression. Results Acacetin significantly promoted the prolifera-tion and increased the amount of cells arrested in S and G2 /M phase under the concentration of 0.001 ~1 0μmol·L -1 .Ki67 mRNA level and the ERαprotein level in T47D cells were remarkably upregulated after acacetin treatment.To clarify which estrogen receptors played a role in acacetin induced the proliferation of T47D cells,the combination treatment of acacetin and ERαinhibitor (MPP)/ERβ inhibitor (PHTPP) was employed.We found that MPP could reverse the cell proliferation,the cell arrested in S and G2 /M phase and the increased Ki67 mRNA level induced by acace-tin.PHTPP also alleviated the T47D cell proliferation induced by acacetin,whereas no significant changes were found in cell cycle and Ki67 mRNA level.Con-clusion Acacetin stimulates the cell proliferation of T47D cells in the concentration from 0.001 μmol · L -1 to 1 0 μmol·L -1 ,which is mainly mediated by ERα.

Expression of estrogen and progesterone receptors in astrocytomas: a literature review

Gliomas are the most common type of primary central nervous system neoplasm. Astrocytomas are the most prevalent type of glioma and these tumors may be influenced by sex steroid hormones. A literature review for the presence of estrogen and progesterone receptors in astrocytomas was conducted in the PubMed database using the following MeSH terms: “estrogen receptor beta” OR “estrogen receptor alpha” OR “estrogen receptor antagonists” OR “progesterone receptors” OR “astrocytoma” OR “glioma” OR “glioblastoma”. Among the 111 articles identified, 13 studies met our inclusion criteria. The majority of reports showed the presence of estrogen and progesterone receptors in astrocytomas. Overall, higher tumor grades were associated with decreased estrogen receptor expression and increased progesterone receptor expression.

Expression and significance of estrogen receptor alpha in renal tissue of IgA nephropathy / 中国医师杂志

Objective To investigate the expression and significance of estrogen receptor alpha in renal tissue of immunoglobulin A (IgA) nephropathy.Methods Fifty renal tissue samples including forty five cases of IgA nephropathy and five cases of normal expression were collected.The expression of estrogen receptor αt (ERα) was detected by immunohistochemistry method,and its relationship with clinical parameters and the degree of glomerular damage were analyzed.Results ERα was located in the glomerular and renal tubules.With the severity of the lesion,the expression of ERα in renal tissue of IgA nephropathy decreased gradually (P =0.001) and its expression was associated with estimated glomerular filtration rate (eGFR),serum creatinine (Scr),and pathologic grade of IgA nephropathy (r =0.876,-0.818,and -0.736,P ＜ 0.05),The expression of ERα was significantly decreased in hypertensive patients with IgA nephropathy,and the difference was statistically significant compared with that in non-hypertensive group (P =0.011).Conclusions The expression of ERα in renal tissue of IgA nephropathy was significantly decreased and there was a correlation between the degree of renal tissue damage,suggesting that ERα might play a role in the development of renal disease.