17β-estradiol配伍DRSP对绝经综合征患者血清FSH及E2调控作用

目的 研究17β-estradiol配伍DRSP治疗绝经综合征患者血清FSH及E2调控作用。方法 选取2013年11月~2016年11月在丽水市人民医院接受治疗的绝经综合征的女性患者共90例。随机将患者分为3组,17β组（30例,单独使用17β-estradiol治疗）、DRSP组（30例,单独使用DRSP治疗）、联合组（30例,17β-estradiol结合DRSP治疗）。使用Kupperman的绝经综合征的评测法,对患者的身体症状量化,治疗后对患者进行比较分析,观察3组患者的治疗有效率情况,患者在治疗前进行的检查项目包含肝肾功能、血尿常规、空腹时血糖含量、卵泡刺激素（follicle-stimulating hormone,FSH）、E2、低密度脂蛋白（low-density lipoprotein,LDL）、总胆固醇（total cholesterol,TC）、心电图、腔内的多普勒超声、乳腺的彩超、高密度脂蛋白（high-density lipoprotein,HDL）等,治疗前按照Kupperman评测表评价一次,在每个疗程结束后再测量一次,并在此进行上述的各项检查。结果 联合组患者在治疗后,除了皮肤上的蚁走感和治疗前没有差异以外,其他的症状的积分差异均都统计学的意义,其中性交痛、头痛、身体感觉异常、容易激动、心悸、关节痛、疲乏、失眠和潮热的症状评分差异显著（P〈0.01）,DRSP组患者在治疗后,性交痛、抑郁和失眠有所改善,差异有统计学意义（P〈0.05）,其他症状均无明显改善;17β组患者在治疗后,头痛、关节疼痛和失眠有所改善,差异有统计学意义（P〈0.05）,其他症状均无明显改善;联合组的总有效率为90.0%,DRSP组的总有效率为73.33%,17β组的总有效率为70.0%,联合组与另外量组对比差异显著（P〈0.05）,治疗前后联合组患者在血TC、E2和FSH对比中差异显著（P〈0.05）,DRSP组患者FSH在治疗前后有差异,其他均无差异,17β组患者E2在治疗前后有差异,其他均无差异。结论 17β-estradiol配伍DRSP治疗绝经综合征患者取得了良好的临床疗效,其体内血清FSH及E2调控作用显著。

17β-estradiol通过促进PSD95/nNOS耦联抑制皮层海马神经元的存活

目的:检测17β-estradiol对皮层海马神经元存活的影响,并判断其与nNOS/PSD95耦联的关系。方法:采用原代培养的ICR小鼠皮层海马神经元,培养7天后分溶剂对照组和给药组分别给予溶剂DMSO和浓度为10μmol/L的17β-estradiol,LDH法判断细胞损伤,测定给药30 min后细胞死亡情况,并拍摄光镜照片观察细胞形态。同时利用免疫共沉淀法检测DMSO组,10nmol/L17β-estradiol组,10μmol/L 17β-estradiol组的nNOS和PSD95的耦联情况,观察药物对其影响。结果:给药30 min后,与对照组相比,10μmol/L17β-estradiol组LDH漏出率增加,同时nNOS/PSD95耦联增加,而10 nmol/L 17β-estradiol对nNOS/PSD95耦联并没有影响。结论:浓度为10μmol/L的17β-estradiol会损伤皮层海马神经元,而且这一结果可能是通过促进nNOS/PSD95耦联实现的。

17β-Estradiol Regulates Cultured Immature Boar Sertoli Cell Proliferation via the cAMP-ERK1/2 Pathway and the Estrogen Receptor β

Estrogen plays an important role in regulating Sertoli cell number in the testis. The objective of the study was to identify whether 17β-estradiol affected the proliferation of cultured, immature boar Sertoli cells via the estrogen receptor β （ERβ） and the cAMP-extracellular signal-regulated kinase （ERK1/2） pathway. Low levels （10-10-10-8 mol L-1） of 17β-estradiol increased cell number, but high levels （10-7-10-6 mol L-1） decreased it （P〈0.05）. Sertoli cell number began to recover for an additional 24 h in the medium without 17β-estradiol （10-6 mol L-l） （P〉0.05）. The effects of 17β-estradiol （10-9 mol L-1） peaked at the first 24 h （P〈0.05）. 17β-estradiol activated ERK1/2 from 5 min to 24 h, but the activiy of ERK1/2 began to decrease after 4 h. Both PD98059 and U0126, two ERK inhibitors, blocked cell division （P〈0.05）. 17β-estradiol （10-10-10-6 mol L-1） dose-dependently increased cAMP production （P 〈 0.05）, and both 17β-estradiol （10-9 mol L-1） and forskolin, which increases cAMP levels, induced cell proliferation and activated ERK1/2 （P〈 0.05）. Rp-cAMP, an antagonist of cAMP, blocked this 17β-estradiol activity （P〈 0.05）. Two estrogen receptor antagonists, ICI 182780 and ERβ antagonist （ERβAnt）, reduced Sertoli cell number, cAMP production and ERK1/2 activation （P〈 0.05）, but ERaAnt did not （P〉 0.05）. Therefore, 17β- estradiol mainly promotes pig Sertoli cell proliferation via ERβ to induce cAMP production and ERK activation to promote cell proliferation.

Xenoestrogens challenge 17β-estradiol protective effects in colon cancer

Several epidemiological,cellular,and molecular studies demonstrate the role of environmental chemicals with endocrine disrupting activities,typical of Westernized societies,in the pathogenesis of numerous diseases including cancer.Nonetheless this information,the design and execution of studies on endocrine disruptors are not yet cognizant that the specific actions of individual hormones often change with development and ageing,they may be different in males and females and may be mediated by different receptors isoforms expressed in different tissues or at different life stages.These statements are particularly true when assessing the hazard of endocrine disruptors against 17β-estradiol(E2)actions in that this hormone is crucial determinant of sexrelated differences in anatomical,physiological,and behavioral traits which characterize male and female physiology.Moreover,E2 is also involved in carcinogenesis.The oncogenic effects of E2 have been investigated extensively in breast and ovarian cancers where hormone-receptor modulators are now an integral part of targeted treatment.Little is known about the E2preventive signalling in colorectal cancer,although this disease is more common in men than women,the difference being more striking amongst pre-menopausal women and age-matched men.This review aims to dissect the role and action mechanisms of E2 in colorectal cancer evaluating the ability of estrogen disruptors(i.e.,xenoestrogens)in impair these E2 actions.Data discussed here lead to define the possible role of xenoestrogens in the impairment and/or activation of E2signals important for colorectal cancer prevention.

Recovery of gonadal development in tiger puffer Takifugu rubripes after exposure to 17β-estradiol during early life stages

The aim of the present study was to investigate the long-term effects of 17l）-estradiol （E2） exposure on gonadal development in the tiger puffer （Taktfugu rubripes）, which has a genetic sex determination system of male homogametic XY-XX. Tiger puffer larvae were exposed to 1, 10 and 100 μg/L E2 from 15 to 100 days post-hatch （dph） and then maintained in clean seawater until 400 dph. Changes in sex ratio, gonadal structure and gonadosomatic index （GSI） were monitored at 100, 160, 270 and 400 dph. Sex-associated single nucleotide polymorphism （SNP） markers were used to analyze the genetic sex of samples, except those at 100 dph. Exposure had a positive effect on the conversion of genetically male gonads into phenotypically female gonads at 100 dph. However, gonads from 60% of genetic XY males in the 1-μg/L E2 group and 100% in the 10-μg/L E2 group developed intersexual gonads at 160 dph; gonads of all genetic XY males in the two treatment groups reverted to testis by 270 dph. While 38%, 57% and 44% of gonads of XY fish in the 100-gg/L E2 group reverted to intersexual gonads at 160, 270 and 400 dph, respectively, none reverted to testis after E2 treatment. In addition, E2 exposure inhibited gonadal growth of both genetic sexes, as indicated by the clear dose-dependent decrease in GSI at 270 and 400 dph. The results showed that exposure to E2 during the early life stages of tiger puffer disrupted gonadal development, but that fish recovered after migration to clean seawater. The study suggests the potential use of tiger puffer as a valuable indicator species to evaluate the effects of environmental estrogens on marine fish, thereby protecting valuable fishery resources.

17β-estradiol inhibits TGF-β-induced collagen gel contraction mediated by human Tenon fibroblasts via Smads and MAPK signaling pathways

AIM:To investigate the impact of 17β-estradiol on the collagen gels contraction(CGC)and inflammation induced by transforming growth factor(TGF)-βin human Tenon fibroblasts(HTFs).METHODS:HTFs were three-dimensionally cultivated in type I collagen-generated gels with or without TGF-β(5 ng/mL),17β-estradiol(12.5 to 100μmol/L),or progesterone(12.5 to 100μmol/L).Then,the collagen gel diameter was determined to assess the contraction,and the development of stress fibers was analyzed using immunofluorescence staining.Immunoblot and gelatin zymography assays were used to analyze matrix metalloproteinases(MMPs)and tissue inhibitors of metalloproteinases(TIMPs)being released into culture supernatants.Enzyme-linked immunosorbent assay(ELISA)and reverse transcription-quantitative polymerase chain reaction(RT-PCR)were used to detect interleukin(IL)-6,monocyte chemoattractant proteins(MCP)-1,and vascular endothelial growth factor(VEGF)in HTFs at the translational and transcriptional levels.The phosphorylation levels of Sma-and Mad-related proteins(Smads),mitogen-activated protein kinases(MAPKs),and protein kinase B(AKT)were measured by immunoblotting.Statistical analysis was performed using either the Tukey-Kramer test or Student’s unpaired t-test to compare the various treatments.RESULTS:The CGC caused by TGF-βin HTFs was significantly inhibited by 17β-estradiol(25 to 100μmol/L),and a statistically significant difference was observed when comparing the normal control group with 17β-estradiol concentrations exceeding 25μmol/L(P<0.05).The suppressive impact of 17β-estradiol became evident 24h after administration and peaked at 72h(P<0.05),whereas progesterone had no impact.Moreover,17β-estradiol attenuated the formation of stress fibers,and the production of MMP-3 and MMP-1 in HTFs stimulated by TGF-β.The expression of MCP-1,IL-6,and VEGF mRNA and protein in HTFs were suppressed by 100μmol/L 17β-estradiol(P<0.01).Additionally,the phosphorylation of Smad2 Smad3,p38,and extracellular signal-regulated kinase(ERK)were downregulated(P<0.01).CONCLUSION:17β-estradiol significantly inhibits the CGC and inflammation caused by TGF-βin HTFs.This inhibition is likely related to the suppression of stress fibers,inhibition of MMPs,and attenuation of Smads and MAPK(ERK and p38)signaling.17β-estradiol may have potential clinical benefits in preventing scar development and inflammation in the conjunctiva.

Relaxation of rabbit cavernous smooth muscle to 17β-estradiol: a non-genomic, NO-independent mechanism

Aim: To investigate whether estrogen was involved in relaxation of rabbit cavernous smooth muscle. Methods: Relaxation response of the rabbit cavernous smooth muscles to 17β-estradiol (0.3, 3, 30 and 300 nmol/L) were observed in vitro. The response of the muscle strips to estrogen after incubation with either actinomycin D (10μmol/L) or L-NAME (10μmol/L) were also evaluated. Inside-out mode of patch clamp in a single smooth muscle cell was applied to investigate the Maxi-K channel activities. Results: Estrogen caused a dose-dependent relaxation of the strips precontracted with norepinephrine. The maximal response was noted about 10 minutes after treatment. The estrogen-induced relaxation was prevented by neither actinomycin D nor L-NAME, suggesting that the response was not mediated by gene transcription or nitric oxide (NO). Application of 17β-estradiol increased the Maxi-K channel activities. Conclusion: 17β-estradiol may be involved in relaxation of rabbit cavernous smooth muscles via a non genomic and NO independent mechanism. 17β-estradiol stimulates Maxi-K channel of the rabbit cavernous myocyte.

Induction of testis-ova in nile tilapia (Oreochromis niloticus) exposed to 17β-estradiol

The efficacy of Endocrine Disrupting Compounds (EDCs), 17β-estradiol was tested on the fish Oreochromis niloticus in order to understand the intersex relationship of fish, in which sequential hermaphrodism can consist of a male changing into a female (protandry) or a female changing into a male (protogyny). The fish were equally divided into 3 groups. The first group was the control group;the second and third groups were treated with 10 and 100 mg L-1 of 17β-estradiol, respectively, for 30 days. The overall result in this experiment had no significant effect on the growth parameters. Among the two treated groups, the low concentration group shows results similar to those of the control groups. The high concentration group shows changes to the male reproductive system with the appearance of the testis-ova present resulting in an intersex condition of the male gonads. With this experiment, it can be concluded that 17β-estradiol at high concentration reveals positive changes towards the male reproductive system of the fish, Oreochromis niloticus.

iTRAQ-based analysis of 17β-estradiol induced proteome in Chinese tongue sole Cynoglossus semilaevis

The phenomenon of sex dimorphism prevails among many fi sh species. It has attracted the general research interest due to its close relationship to fi sh growth and thus aquaculture productivity. In Chinese tongue sole ( Cynoglossus semilaevis ), 17β-estradiol (E2) can be used reportedly to induce the feminization of fi sh, although the detailed regulatory network remained elusive. We treated the tongue sole fry before sex diff erentiation with E2 (10 and 30 μg/L) to identify potential targets of E2 in Chinese tongue sole. The E2 treatment indeed induced genetic male fi sh sex-reversal to phenotypic female. Using an iTRAQ-based comparative proteomic analysis, 409 proteins that diff erentially expressed after E2 induction were identifi ed, including 259 up-regulated and 150 down-regulated proteins. Furthermore, 19 diff erentially expressed proteins identifi ed in the comparative proteomic analysis were selected to assess their transcription and eight of them exhibited the same tendency. Functions of the eight proteins included mainly nucleotide and protein binding. Interestingly, a far upstream element-binding protein 3-like isoform exhibited the signifi cant upregulation both at transcription and translation levels after E2 treatment. This work identifi ed a set of candidate proteins for E2 response and deepened our understanding of E2 regulatory mechanism.

Corrigendum to: Expression profiles of sex-related genes in gonads of genetic male Takifugu rubripes after 17β-estradiol immersion

The original version of this article unfortunately contained a mistake. The publication year in the header on the first page (p.1113) of this article was incorrect. The corrected publication year is given below:

17β-Estradiol Regulates SKP2 Expression in Cultured Immature Boar Sertoli Cells Mainly via Estrogen Receptor β,cAMP-PKA and ERK1/2

Estrogen plays an important role in regulating testicular Sertoli cell number. Furthermore, S-phase kinase-associated protein 2 （SKP2） plays a central role in mammalian cell cycle progression. The objective of this study was to determine whether 17β-estradiol can regulate the expression of SKP2, and the Sertoli cell cycle, via estrogen receptor β （ERβ）, the cyclic adenosine monophosphate （cAMP）-protein kinase A （PKA） and extracellular signal-regulated kinase （ERK1/2） pathway. When cultured immature boar Sertoli cells were treated with 17β-estradiol, a time-dependent increase in SKP2 mRNA and protein level was observed by real-time PCR and Western blot, and 17β-estradiol activity peaked at 30 min. Treatment with ICI182780 and ERβ antagonist reduced 17β-estradiol-induced expression of SKP2 and proliferating cell nuclear antigen （PCNA）, while increasing the protein concentration of p27kip1. However, the effect of ERa antagonist on these parameters was lower than that of ICI 182780 and ERβ. Forskolin had a similar effect as 17β-estradiol on the expression of SKP2, PCNA and p27kip1, Rp-cAMP, H-89 and U0126 treatment reduced 17β-estradiol-induced changes, while H-89 also inhibited ERK1/2 activation. Therefore, 17β-estradiol mainly regulates SKP2 mRNA and protein expression via ERβ-cAMP-PKA and ERK1/2 activation. SKP2 and PCNA expression were positively correlated, while increased SKP2 expression likely resulted in p27kip1 degradation.

Effects of Ovariectomy and 17β-Estradiol Replacement on the Activity of Dopamine D2 Receptors in the Selection of Macronutrients Carbohydrates, Lipids and Proteins in Females Rats

17β-estradiol modulates the activity of D2 receptors in the regulation of food intake and body weight. The functional lack of 17β-estradiol in postmenopausal women could create a dietary imbalance and cause body weight gain. This study aimed to better understand the interferences that could exist between 17β-estradiol, D2 receptors and the selection of carbohydrate, fat and protein consumption, as well as their consequences on body weight gain by using an animal model of the menopause. Ovariectomy exacerbates the consumption of foods rich in lipids. Thus confirming an inhibitory action of 17β-estradiol (E2) on the consumption of these types of foods. This consumption stimulates body weight gain, which is promoted by the high caloric content of these foods and not by the amount consumed. Our results showed a direct involvement of D2 receptors in food choice. This choice would be made according to the two (2) isoforms of the D2 receptors. The D2/BR isoform directs towards a high carbohydrate consumption, without causing a gain in body weight. While D2/SUL, promotes high fat food consumption, causing an increase in body weight. In women, 17β-estradiol modulates the activity ratio between these two D2 receptor isoforms to ensure energy and homeostatic balance, stabilizing food intake and body weight.

Serum 17β-estradiol and oral dryness feeling in menopause

The aim of this study was to compare serum 17β-estradiol of menopausal women with/without Oral Dryness (OD) feeling, and evaluate the re-lationship between serum 17β-estradiol and severity of OD feeling. A case-control study was carried out on 70 selected menopausal women aged 40 - 77 years with or without OD feeling (35 as case, 35 as control) conducted at the Clinic of Oral Medicine, Tehran University of Medical Sciences. Xerostomia inventory (XI) score was used as an index of OD feeling severity. The serum 17β-estradiol concentration was measured by an enzyme immunoassay kit (ELISA). Statistical analysis of Student’s t-test and Spearman correlation coefficient was used. The mean serum concentration of 17β-estradiol was significantly lower in case than control. There was a significant negative correlation between XI score and concentration of 17β-estradiol in menopausal women (r = –0.311, P = 0.004). It seems that there is a negatively slight correlation between OD feeling severity and serum 17β-estradiol in menopausal women.

Impact of 17β-Estradiol on Natural Water’s Heterotrophic Nitrifying Bacteria

In this research, bottom water samples were collected from nature water. After cultivating and selecting, bacteria which could use (NH4)2SO4 as the only nitrogen source had been selected. The bacteria were cultivated in BM cultures with 0, 0.1, 1, 10, 100 ng/L 17β-estradiol (E2), and the initial concentration of E2 is the only difference between cultures of each group. BM culture is a kind of bacteria culture with 100 mg/L of NH4-N as only nitrogen source. Every group’s N- NH4+, N- NO3-concentration and OD600 were measured. The result shows that compared with the control group, in which no E2 was added, the growth of heterotrophic nitrifying bacteria had been promoted when the concentration of E2 was in range of 1 - 100 ng/L. In addition, heterotrophic nitrifying bacteria’s growing speed has a positive correlation between the E2’s concentration. However, low concentration of E2 (like 0.1 ng/L), could inhibit the growth of heterotrophic nitrifying bacteria. Considering the impact of E2 on heterotrophic nitrifying bacteria, it is necessary to intensify the detection of E2 in the future.

Removal of 17β-Estradiol by Electro-Fenton Process

The Electrochemical advanced oxidation method “Electro-Fenton” has been applied to remove 17β-estradiol (17β)- estra-1,3,5(10)-triéne-3,17-diol) in aqueous-acetonitrile mixture. This endocrine disrupting is a steroid hormone, releases from humans, animals and residual pharmaceuticals into the environmental water and usually causes suspected undesirable effects in aquatic organisms. The degradation of this organic compound by Electro-Fenton process was showed using a carbon felt cathode and platinum anode. The evolution of the concentration during treatment was followed up by high performance liquid chromatography (HPLC). The influence of operating conditions on the degradation of 17β-estradiol by Electro-Fenton step, such as initial concentration and catalyst concentration, has been investi- gated and discussed. We showed that the degradation reaction obeyed apparent first-order reaction kinetics, with absolute rate constant determined as 5.12 × 109 M–1 s–1 by competitive kinetics method taking Benzoic Acid as reference compound. The results confirm the efficiency of the Electro-Fenton process to degrade organic pollutant in aqueous-acetonitrile mixture.

Helicobacter pylori and 17β-estradiol induce human intrahepatic biliary epithelial cell abnormal proliferation and oxidative DNA damage

BACKGROUND: Biliary cancers are more common in females, and previous studies have suggested that Helicobacter pylori (H. pylori) exists in the biliary system. However, the effects of H. pylori infection and estrogen on the biological behaviors of human biliary epithelium mucosa remain unknown. The present study aimed to clarify their effects on the proliferation, apoptosis, migration and oxidative DNA damage of a human intrahepatic biliary epithelial cell (HIBEC) line in vitro. METHODS: HIBECs were co -cultured with 17 beta-estradiol (at 10(-9) mol/L, 10(-7) mol/L, and 10(-5) mol/L) and H. pylori (at MOI=0.5:1, 1:1, and 2:1) and continuously passaged until the 15th generation (approximately 45 days). Then, the following assays were performed. HIBEC proliferation was measured using the CCK-8 assay, plate clone-formation assay and by determining Ki-67 expression with immunocytochemistry; cell apoptosis and migration were investigated using Annexin-V/PI and transwell assays, respectively; and reactive oxygen species (ROS) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) production were detected by flow cytometry and immunofluorescence staining combined with confocal laser scanning microscopy, respectively. The results were the basis for evaluating the level of oxidative stress and the related DNA damage in HIBECs. RESULTS: HIBECs maintained a normal morphology and vitality when treated with 17 beta-estradiol (at 10(-9) mol/L) and H. pylori (at MOI=0.5:1 and 1:1). 17 beta-estradiol at 10(-7) mol/L and 10(-5) mol/L and H. pylori at MOI=2:1, by contrast, caused cell death. Compared with controls, HIBECs treated with 17 beta-estradiol (10(-9) mol/L) and H. pylori (MOI=1:1) had a higher up-regulation of proliferation, Ki-67 expression, clone formation, migration activity and the expression of ROS and 8-OHdG and exhibited a down-regulation of apoptosis. The above effects were further increased when 17 beta-estradiol and H. pylori were combined (P<0.05). CONCLUSIONS: H. pylori and 17 beta-estradiol, separately or in combination, promoted cell proliferation and suppressed apoptosis of HIBECs in vitro. The above phenomena might be related to oxidative stress and its subsequent DNA damage with H. pylori and 17 beta-estradiol.

Expression of Nkx3.1 enhances 17β-estradiol anti-tumor action in PC3 human prostate cancer cells

Aim： To explore whether the anti-tumor action of 17β-estradiol is enhanced by re-expression of the homeodomain transcription factor Nkx3.1 in PC3 human prostate cancer cells. Methods： PC3 cells were stably transfected with pcDNA3.1-Nkx3.1-His vector, which carries a full-length cDNA of human Nkx3.1. The PC3 cells stably transfected with vector pcDNA3.1 were set as a control. The expression of Nkx3.1 protein in the cells was confirmed by Western blot analysis. The effect of Nkx3.1 on cell proliferation of PC3 cells was examined with MTT assay. The antiproliferative and apoptotic effects of 17β-estradiol alone or in combination with Nkx3.1 were estimated on PC3 cells by using MTT growth tests and flow cytometric analyses. The expression of apoptosis-related proteins was analyzed using Western blotting. Results： The plasmid carrying Nkx3.1 gene induced high expression of Nkx3.1 protein in PC3 cells. The re-expression of exogenous Nkx3.1 did not cause a significant reduction in cellular proliferation, whereas the expression of Nkx3.1 enhanced the 17β-estradiol anti-proliferative effect in PC3 cells. Nkx3.1 expres- sion promoted 17β-estradiol-induced apoptosis of PC3 cells, as shown by analysis of Bcl-2, Bax, Caspase-3 and poly （ADP-ribose） polymerase expression. Conclusion： The present study demonstrates that re-expression of Nkx3.1 enhances 17β-estradiol anti-tumor action in PC3 human prostate cancer cells. The in vitro study suggests that re- expression of Nkx3.1 is worthy of further consideration as an adjuvant treatment of androgen independent prostate cancer with estrogen anti-tumor therapies.

ASSESSMENT OF 17β-ESTRADIOL IN AQUATIC ENVIRONMENTS AROUND KLANG VALLEY,MALAYSIA

This study was conducted to assess the existing concentration of 17β-estradiol(E2)in the surface water samples collected from rivers and lakes around Klang Valley,Malaysia.E2,which is a natural feminizing chemical produced in female organisms,regularly used to compare with other environmental estrogens because they behave similarly and react effectively as a hormone at a very low concentration.It was found that the average concentration of E2 in the aquatic environment of Klang Valley was(14.08 ±3.67)pg/mL,which was 14 times higher than those in the Japanese aquatic environment in this study.The river system had the average concentration of(20.02±5.26)pg/mL while the lake had an average concentration of(5.91±3.39)pg/mL.The E2 concentration was presumed high if the sources occurred nearby the area.Current levels of E2 in the aquatic environment may possess threats to existing aquatic organisms.Since high level of E2 has been discovered in the aquatic environment around Klang Valley,further studies and monitoring of E2 and other EDCs concentrations are needed to determine their levels in Malaysian aquatic environment and help to control these chemicals pollution in the aquatic environment.

Suppressive effects of 17β-estradiol on hepaUc fibrosis in CCl_4-induced rat model

AIM: To investigate the pathway via which 17β-estradio(β-Est) exerts suppressive effects on rat hepatic fibrosis.METHODS: In vivo study was done in CCI4-induced female hepatofibrotic rats. Fibrosis-suppressive effect of β-Est(20 μg/kg·d) was evaluated in intact and ovariectomized rat models. Six weeks after the treatment, all the rats were sacrificed and specimens of serum or liver tissue were collected for the studies. Serum liver enzymes,fibrosis markers and estradiol levels were determined by standard enzymatic methods, ELISA and RIA, respectively.Degrees of fibrosis and areas of hepatic stellate cells(HSCs) positive for alpha-smooth muscle actin (a-SMA) in the liver were determined by van Gieson (VG) stain and immunohistochemistry. In vitro studies, HSCs were isolated by a combination of pronase-collagenase perfusion and density gradient centrifugation. First-passage HSCs were randomly divided into 10 groups, and different concentrations of β-Est, 2-hydroxyestradiol (2OHE) or 2-methoxyestradiol(2MeOE) were separately added to the cell groups. After incubation for 72 h, the degree of cell proliferation, collagen production, ^-SMA or estrogen receptor (ER) expression was determined by MTT assay, ELISA and immunohistochemistry,respectively.RESULTS: β-Est treatment reduced aspartate aminotransfer-ase (AST), alanine aminotransferase (ALT), hyaluronic acid(HA) and type IV collagen (C IV) in sera, suppressed hepatic collagen content, decreased the areas of HSCs positive for α-SMA significantly in both intact and ovariectomized female hepatofibrotic rats. There was a negative correlation between the percentage of fibrotic area of liver tissue and the serum estradiol level; the calculated correlation coefficient was -0.57 (P<0.01). β-Est and its metabolites concentration-dependently (10^-9 mol/L-10^-7 mol/L) inhibited HSC proliferation and collagen synthesis. At the concentration of 10^-7 mol/L, they could inhibit α-SMA expression. The order of potency was 2MeOE>2OHE>β-Est.CONCLUSION: β-Est may suppress hepatic fibrosis probably via its biologically active metabolites.

17β-Estradiol,through activating the G protein-coupled estrogen receptor,exacerbates the complication of benign prostatic hyperplasia in type 2 diabetes mellitus patients by inducing prostate proliferation

Benign prostatic hyperplasia(BPH)is one of the major chronic complications of type 2 diabetes mellitus(T2DM),and sex steroid hormones are common risk factors for the occurrence of T2DM and BPH.The profiles of sex steroid hormones are simultaneously quantified by LC-MS/MS in the clinical serum of patients,including simple BPH patients,newly diagnosed T2DM patients,T2DM complicated with BPH patients and matched healthy individuals.The G protein-coupled estrogen receptor(GPER)inhibitor G15,GPER knockdown lentivirus,the YAP1 inhibitor verteporfin,YAP1 knockdown/overexpression lentivirus,targeted metabolomics analysis,and Co-IP assays are used to investigate the molecular mechanisms of the disrupted sex steroid hormones homeostasis in the pathological process of T2DM complicated with BPH.The homeostasis of sex steroid hormone is disrupted in the serum of patients,accompanying with the proliferated prostatic epithelial cells(PECs).The sex steroid hormone metabolic profiles of T2DM patients complicated with BPH have the greatest degrees of separation from those of healthy individuals.Elevated 17β-estradiol(E2)is the key contributor to the disrupted sex steroid hormone homeostasis,and is significantly positively related to the clinical characteristics of T2DM patients complicated with BPH.Activating GPER by E2 via Hippo-YAP1 signaling exacerbates high glucose(HG)-induced PECs proliferation through the formation of the YAP1-TEAD4 heterodimer.Knockdown or inhibition of GPER-mediated Hippo-YAP1 signaling suppresses PECs proliferation in HG and E2 co-treated BPH-1 cells.The anti-proliferative effects of verteporfin,an inhibitor of YAP1,are blocked by YAP1 overexpression in HG and E2 co-treated BPH-1 cells.Inactivating E2/GPER/Hippo/YAP1 signaling may be effective at delaying the progression of T2DM complicated with BPH by inhibiting PECs proliferation.