Rabbit Anti-Human PDIA6 (a.a. 1-440) Polyclonal antibody

The abuse of anabolic androgenic steroids (AAS) has been considered a major public health problem during decades. Supraphysiological doses of AAS may lead to a variety of neuroendocrine problems. Precisely, the hypothalamic-pituitary-gonadal (HPG) axis is one of the body systems that is mainly influenced by steroidal hormones. Fluctuations of the hormonal milieu result in alterations of reproductive function, which are made through changes in hypothalamic neurons expressing gonadotropin-releasing hormone (GnRH). In fact, previous studies have shown that AAS modulate the activity of these neurons through steroid-sensitive afferents. To increase knowledge about the cellular mechanisms induced by AAS in GnRH neurons, we performed proteomic analyses of the murine hypothalamic GT1-7 cell line after exposure to 17a-methyltestosterone (17 alpha-meT; 1 mu M). These cells represent a good model for studying regulatory processes because they exhibit the typical characteristics of GnRH neurons, and respond to compounds that modulate GnRH in vivo. Two-dimensional difference in gel electrophoresis (2D-DIGE) and mass spectrometry analyses identified a total of 17 different proteins that were significantly affected by supraphysiological levels of AAS. Furthermore, pathway analyses showed that modulated proteins were mainly associated to glucose metabolism, drug detoxification, stress response and cell cycle. Validation of many of these proteins, such as GSTM1, ERH, GAPDH, PEBP1 and PDIA6, were confirmed by western blotting. We further demonstrated that AAS exposure decreased expression of estrogen receptors and GnRH, while two important signaling pathway proteins p-ERK, and p-p38, were modulated. Our results suggest that steroids have the capacity to directly affect the neuroendocrine system by modulating key cellular processes for the control of reproductive function.

In our recent study, we found that the expression levels of total xylose-binding proteins (XBPs) were up-regulated significantly in activated hepatic stellate cells (HSCs); however, the denomination, distribution, and function of the XBPs were uncharted. Herein, 70 XBPs from activated HSCs and 64 XBPs from quiescent HSCs were isolated, identified and annotated. A total of 30 XBPs were up-regulated (all fold change >= 1.5, p <= 0.05) and 14 XBPs were down-regulated (all fold change <= 0.67, p <= 0.05) in the activated HSCs. The XBPs were localized at the cytoplasm and cytoplasmic membrane in HSCs and cirrhotic liver tissues by cy/histochemistry. The XBPs (i.e. PDIA6 and CFL2) responsible for the regulation of protein binding were up-regulated and those responsible for the regulation of catalytic activity (i.e. TUBB and MX1) were up-regulated in the activated HSCs. 2 candidates (i.e. PDIA6 and APOA1) were then selected for further verification in the sera of patients with HBV-induced chronic hepatitis/cirrhosis using western blotting and serum microarrays. PDIA6 showed a higher discrimination (Area Under Curves, AUCs = 0.8985, p < 0.0001) relative to APOA1 (AUCs = 0.8738, p < 0.0001) in the sera of patients as biomarker candidate. In conclusion, the precision alteration of the XBPs associated with pathological changes in HSCs during liver fibrosis/cirrhosis may provide pivotal information needed to discover potential glycan-binding protein-related biomarkers for diagnosis of liver fibrosis/cirrhosis and for development of new anti-fibrotic strategies.