Avian SERPINB12 Expression in the Avian Oviduct Is Regulated by Estrogen and Up-Regulated in Epithelial Cell-Derived Ovarian Carcinomas of Laying Hens

Serine protease inhibitors (SERPINs) are involved in a variety of biological processes such as blood clotting, angiogenesis, immune system, and embryogenesis. Although, of these, SERPINB12 is identified as the latest member of clade B in humans, little is known of it in chickens. Thus, in this study, we investigated SERPINB12 expression profiles in various tissues of chickens and focused on effects of steroid hormone regulation of its expression. In the chicken oviduct, SERPINB12 mRNA and protein are abundant in the luminal (LE) and glandular (GE) epithelia of the magnum in response to endogenous or exogenous estrogen. Furthermore, SERPINB12 mRNA and protein increase significantly in GE of cancerous ovaries of laying hens with epithelia-derived ovarian cancer. Collectively, these results indicate that SERPINB12 is a novel estrogen-stimulated gene that is up-regulated by estrogen in epithelial cells of the chicken oviduct and that it is a potential biomarker for early detection of ovarian carcinomas in laying hens and women

Berberine Improves Benign Prostatic Hyperplasia via Suppression of 5 Alpha Reductase and Extracellular Signal-Regulated Kinase in Vivo and in Vitro

Benign prostate hyperplasia (BPH) is a common disease in elderly men, characterized by proliferated prostate and urinary tract symptoms. The hormonal cascade starting by the action of 5-alpha-reductase (5AR) is known to be one of the pathways responsible for the pathogenesis of BPH. Present investigation evaluated the capacity of berberine (BBR), a nature-derived compound abundant in Coptis japonica, in testosterone-induced BPH rats. Experimental BPH was induced by inguinal injection with testosterone propionate (TP) for 4 weeks. BBR or finasteride, a 5AR inhibitor as positive control, was treated for 4 weeks during BPH. BPH induced by TP evoked weight gaining and histological changes of prostate and BBR treatment improved all the detrimental effects not only weight reduction and histological changes but also suppression of prostate-specific antigen (PSA), which is elevated during BPH. Additionally, BBR suppressed TP-associated increase of 5AR, androgen receptor (AR) and steroid coactivator-1 (SRC-1), the key factors in the pathogenesis of BPH. To evaluate the underlying molecular mechanisms responsible for beneficial effects of BBR, we investigated whether these effects were associated with the mitogen-activated protein kinase pathway. BPH induced by TP showed increased phosphorylation of extracellular signal-regulated kinase (ERK), whereas this was suppressed by BBR treatment. On the other hand, c-jun-N-terminal kinase (JNK) and p38 mitogen-activated protein kinase was not changed in BPH rats. In in vitro study using RWPE-1 cells, a human prostate epithelial cell line. TP increased cell proliferation and BPH-related key factors such as PSA, AR, and 5AR in RWPE-1 cells, and those factors were significantly decreased in the presence of BBR. Furthermore, these proliferative effects in RWPE-1cells were attenuated by treatment with U0126, an ERK inhibitor, confirming BBR can relieve overgrowth of prostate via ERK-dependent signaling. The cotreatment of U0126 and BBR did not affect the change of 5AR nor proliferation compared with U0126 alone, suggesting that the effect of BBR was dependent on the action of ERK. In conclusion, this study shows that BBR can be used as a therapeutic agent for BPH by controlling hyperplasia of prostate through suppression of ERK mechanism

Expression of SERPINB12 in various organs of both male and female chickens.

<p>Results of RT-PCR analysis using cDNA templates from different organs of both male [A] and female [B] chickens with chicken SERPINB12 and chicken GAPDH-specific primers are shown. SERPINB12 gene was generally expressed in all organs from male and female chickens. Its expression was highly abundant in the chicken oviduct.</p

Quantitation of SERPINB12 mRNA during regression and recrudescence of the magnum of laying hens during induced molting.

<p>Quantitative PCR was conducted using cDNA templates from the magnum of hens fed a control diet (Day 0), zinc fed hens in which the oviduct was regressing (Day 6 and 12) [A], and hens in which the oviduct was undergoing recrudescence (Day 20, 25, 30, and 35) [B]. These experiments were performed in triplicate and normalized to control <i>GAPDH</i> expression. The asterisks denote statistically significant differences (mean ± SEM; ** <i>P</i><0.01 or *** <i>P</i><0.001).</p

Localization of SERPINB12 mRNA and protein in oviducts of DES-treated and non-treated chicks.

<p>[A] <i>In situ</i> hybridization analyses indicated cell-specific expression of <i>SERPINB12</i> mRNA in GE and LE of the four segments of oviducts from DES-treated chicks. However, there was no expression of <i>SERPINB12</i> in oviducts from control chicks. [B] Immunoreactive SERPINB12 protein was localized to LE and GE of oviducts of chicks treated with DES, especially the magnum. For the IgG control, normal rabbit IgG was substituted for the primary antibody. Sections were not counterstained. Legend: LE, luminal epithelium; GE, glandular epithelium; <i>Scale bar</i> represents 200 µm (the first columnar panels, sense and IgG) and 50 µm (the second columnar panels).</p

Cell-specific localization of SERPINB12 mRNA and protein during regression and recrudescence of the magnum of oviducts from hens during induced molting.

<p>The localization of <i>SERPINB12</i> mRNA in the magnum of hens on different days of the regression phase [A] and recrudescence phase [B] was analyzed by <i>in situ</i> hybridization. It was predominantly expressed in GE and LE of the magnum on Days 0, 25, 30 and 35 of the recrudescence phase of molting. On the other hand, the expression of SERPINB12 was rarely detected on Days 6, 12 and 20 of the regression phase of dietary zinc induced molting. Immunoreactive SERPINB12 protein in the magnum during the regression phase [C] and recrudescence phase [D] was analyzed by immunohistochemistry. Consistent with <i>SERPINB12</i> mRNA localization, SEPRINB12 protein was mainly detected in GE and LE of the magnum on Days 0, 25, 30 and 35 during the recrudescence phase of the oviduct during molting. Legend: LE, luminal epithelium; GE, glandular epithelium; <i>Scale bar</i> represents 200 µm (the first horizontal panels, sense and IgG) and 50 µm (the second horizontal panels).</p

Raphani Semen (Raphanus sativus L.) Ameliorates Alcoholic Fatty Liver Disease by Regulating De Novo Lipogenesis

In this study, we investigated the pharmacological effect of a water extract of Raphani Semen (RSWE) on alcoholic fatty liver disease (AFLD) using ethanol-induced AFLD mice (the NIAAA model) and palmitic acid (PA)-induced steatosis HepG2 cells. An RSWE supplement improved serum and hepatic triglyceride (TG) levels of AFLD mice, as well as their liver histological structure. To explore the molecular action of RSWE in the improvement of AFLD, we investigated the effect of RSWE on four major pathways for lipid homeostasis in the liver: free fatty acid transport, lipogenesis, lipolysis, and &beta;-oxidation. Importantly, RSWE decreased the mRNA expression of de novo lipogenesis-related genes, such as Srebf1, Cebpa, Pparg, and Lpin1, as well as the protein levels of these factors, in the liver of AFLD mice. That these actions of RSWE affect lipogenesis was confirmed using PA-induced steatosis HepG2 cells. Overall, our findings suggest that RSWE has the potential for improvement of AFLD by inhibiting de novo lipogenesis

Distribution and localization of SERPINB12 mRNA and protein in normal and cancerous ovaries from laying hens.

<p>[A] Quantitative PCR was conducted using cDNA templates from normal and cancerous ovaries of hens. The expression of <i>SERPINB12</i> was unique to cancerous ovaries of laying hens as compared with normal ovaries from laying hens. [B] <i>In situ</i> hybridization analyses indicated cell-specific localization of <i>SERPINB12</i> mRNA in normal and cancerous ovaries from laying hens. <i>SERPINB12</i> was strongly expressed in the GE of cancerous ovaries whereas there was no expression in any cells of normal ovaries. [C] Immunoreactive SERPINB12 protein, as for <i>SERPINB12</i> mRNA, was also unique to the GE of cancerous ovaries from laying hens. Legend: F, follicle; GE, glandular epithelium. <i>Scale bar</i> represents 50 µm (the first columnar panels, sense and IgG) and 200 µm (the second columnar panels).</p