Selective estrogen receptor modulator BC-1 activates antioxidant signaling pathway in vitro via formation of reactive metabolites.

AIM: Benzothiophene compounds are selective estrogen receptor modulators (SERMs), which are recently found to activate antioxidant signaling. In this study the molecular mechanisms of antioxidant signaling activation by benzothiophene compound BC-1 were investigated. METHODS: HepG2 cells were stably transfected with antioxidant response element (ARE)-luciferase reporter (HepG2-ARE cells). The expression of nuclear factor erythroid 2-related factor 2 (Nrf2) in HepG2-ARE cells was suppressed using siRNA. The metabolites of BC-1 in rat liver microsome incubation were analyzed using LC-UV and LC-MS. RESULTS: Addition of BC-1 (5 µmol/L) in HepG2-ARE cells resulted in a 17-fold increase of ARE-luciferase activity. Pretreatment with the estrogen receptor agonist E2 (5 µmol/L) or antagonist ICI 182,780 (5 µmol/L) did not affect BC-1-induced ARE-luciferase activity. However, transfection of the cells with anti-Nrf2 siRNA suppressed this effect by 79%. Addition of BC-1 in rat microsome incubation resulted in formation of di-quinone methides and o-quinones, followed by formation of GSH conjugates. BC-1 analogues with hydrogen (BC-2) or fluorine (BC-3) at the 4' position did not form the di-quinone methides. Both BC-2 and BC-3 showed comparable estrogenic activity with BC-1, but did not induce ARE-luciferase activity in HepG2-ARE cells. CONCLUSION: Benzothiophene compound BC-1 activates ARE signaling via reactive metabolite formation that is independent of estrogen receptors.

Low NRF2 mRNA expression in spermatozoa from men with low sperm motility.

Antioxidant genes and enzymes play important roles in human spermatogenesis. Although low levels of antioxidant enzyme expression are associated with poor sperm quality, it is not clear whether mRNA expression of antioxidant genes is lower in these men than in normozoospermic men. In this study, 55 asthenozoospermic and oligoasthenozoospermic patients and 65 controls were recruited. Quantitative real-time reverse transcription PCR was performed and the abundance of mRNA of four antioxidant genes known to be important to spermatogenesis were evaluated. These genes were nuclear factor erythroid 2-related factor 2 (NRF2), catalase (CAT), glutathione S-transferase Mu 1 (GSTM1), and superoxide dismutase isoenzyme 2 (SOD2). Results showed the level of NRF2 mRNA expression to be significantly lower in patients than in controls (P < 0.001), but no statistically significant difference in the level of SOD2, CAT, or GSTM1 gene expression was observed between the two groups. A significant correlation was observed between the level of NRF2 mRNA expression and specific sperm function parameters, including concentration, progressive motility, immotility, vitality, and morphology (all P < 0.01). NRF2 expression was also found to be associated with seminal SOD activity and mRNA levels of the CAT and SOD2 genes (all P < 0.05). Therefore, our data demonstrated that the level of NRF2 mRNA expression is significantly lower in human males with low sperm motility and correlated with specific sperm function parameters. This suggests that NRF2 is important to spermatogenesis and may serve as a useful biomarker in the prediction of male infertility.

[Atractylenolide I Can Induce Apoptosis of U266 Cells and Enhance Bortezomib Effect].

OBJECTIVE: To investigate the effect of atractylenolide I on proliferation and apoptosis of U266 cells, and anti-multiple myeloma effect of bortezomib. METHODS: Bortezomib, bortezomib combined atractylenolide I and atractylenolide I at different concentrations were added into U266 cells respectively, cellular proliferation toxicity was evaluated by CCK-8 assay, apoptosis and cell cycle were detected by using flow cytometry with Annexin V-FITC/PI staining. RT-PCR and Western blot analysis were used to detect the mRNA and protein levels of targeting gene Caspase-3,Caspase-9,BCL-2,BAX,JAK2,STAT3 and IL-6, respectively. RESULTS: The proliferation of U266 cells could inhibited by atractylenolide I, and the apoptosis of U266 cells could be promoted by atractylenolide I, also, which showed a dose-dependent manner(P<0.00; r=0.99). Moreover, the atractylenolide I could regulat the mitochondrial pathway(P<0.01). The combination of 2 drugs could strengther the inhibition of U266 cell proliferation significantly, and the expression level of IL-6,JAK2,STAT3 and BCL-2 mRNA and protein could be decreased by single drug and 2 drugs both(P<0.01). CONCLUSION: Atractylenolide I significantly inhibits the proliferation of U266 cells and promotes their apoptosis. At the same time, it acts synergistically with bortezomib, which may be related to mitochondrial pathway, and probably related to the regulating of IL-6, JAK2 and STAT3 gene expression in signal pathway of JAK2/STAT3.

CR-Unet: A Composite Network for Ovary and Follicle Segmentation in Ultrasound Images.

Transvaginal ultrasound (TVUS) is widely used in infertility treatment. The size and shape of the ovary and follicles must be measured manually for assessing their physiological status by sonographers. However, this process is extremely time-consuming and operator-dependent. In this study, we propose a novel composite network, namely CR-Unet, to simultaneously segment the ovary and follicles in TVUS. The CR-Unet incorporates the spatial recurrent neural network (RNN) into a plain U-Net. It can effectively learn multi-scale and long-range spatial contexts to combat the challenges of this task, such as the poor image quality, low contrast, boundary ambiguity, and complex anatomy shapes. We further adopt deep supervision strategy to make model training more effective and efficient. In addition, self-supervision is employed to iteratively refine the segmentation results. Experiments on 3204 TVUS images from 219 patients demonstrate the proposed method achieved the best segmentation performance compared to other state-of-the-art methods for both the ovary and follicles, with a Dice Similarity Coefficient (DSC) of 0.912 and 0.858, respectively.

A subpathway-based method of drug reposition for polycystic ovary syndrome.

The need for development of new therapeutic agents for polycystic ovary syndrome (PCOS) is urgent due to general lack of efficient and specialized drugs currently available. We aimed to explore the metabolic mechanism of PCOS and inferred drug reposition for PCOS by a subpathway-based method. Using the GSE34526 microarray data from the Gene Expression Omnibus database, we first identified the differentially expressed genes (DEGs) between PCOS and normal samples. Then, we identified 13 significantly enriched metabolic subpathways that may be involved in the development of PCOS. Finally, by an integrated analysis of PCOS-involved subpathways and drug-affected subpathways, we identified 54 novel small molecular drugs capable to target the PCOS-involved subpathways. We also mapped the DEGs of PCOS and a potential novel drug (alprostadil) into purine metabolism pathway to illustrate the potentially active mechanism of alprostadil on PCOS. Candidate agents identified by our approach may provide insights into a novel therapy approach for PCOS.

The effects of cigarette smoke extract on ovulation, oocyte morphology and ovarian gene expression in mice.

Cigarette smoking can harm fertility, but the existing research has targeted primarily on ovarian follicles, embryos or sex hormone. In this study, we tested cigarette smoke extract on ovulation, oocyte morphology and ovarian gene expression associated with inhibition of oxidative stress using C57BL/6 mice. Mice in the experimental group were administered a cigarette smoke extract (CSE) solution (2 mg/ml) orally daily, while the blank control group was given dimethylsulfoxide (DMSO). A positive control group (menadione) was used that received an intraperitoneal injection of 15 mg/kg menadione in oil solution daily. We found that the CSE group manifested a reduced diameter of zona pellucida-free oocyte (ZP-free OD) and a morphologically misshapen first polar body (PB). Our results suggest that CSE exposure is associated with a shrink size and poor quality of oocytes. Quitting smoking is a wise choice to ensure good fertility.