The effect of heat stress on the epidermal growth factor (EGF)-mediated intracellular signaling, and changes cell behavior on swine testicular cell.

At present, high environmental temperature is the main factor endangering animal production, growth and development. Therefore, the harmful effects of heat stress led by hot environment on livestock have attracted much attention. In this work, the cellar property and signaling property of epidermal growth factor (EGF) below heat stress remains unclear in swine testicular cells. Here, the effect of heat stress on EGF-induced intracellular signaling and cell behavior was explored in the ST (a porcine testis cell line). A series of experiments were done to study the cellular behavior and signaling properties of EGF under heat stress. It can be discovered which high ambient temperature changed the cellular characteristics of EGF/EGFR. The eventuates displayed when cells were exposed to thermal environment, EGF/EGFR basically did not internalize, mainly gathered on the cell membrane. Our group also researched the effect of EGF's signaling properties, and the results showed that the ability of EGF to activate EGFR-mediated intracellular signaling decreased significantly under heat stress. Finally, this study illustrated that EGF's cell behavior and signaling profile are obviously altered, indicating that heat stress seriously affected the biological activity of EGF/EGFR, which establish a solid foundation for studying the effect of the EGF on testicular tissue under heat environment.

Beta-carotene regulates the biological activity of EGF in IEC6 cells by alleviating the inflammatory process.

Epidermal growth factor (EGF) has many important biological functions. It plays an important role in regulating the growth, survival, migration, apoptosis, proliferation, and differentiation of intestinal tissues and cells. However, until now, the effect of inflammation on the biological activity of EGF in intestinal cells or tissues is still unclear. For this reason, in the current research, we have conducted a detailed study on this issue. Using the rat small intestinal crypt epithelial cell line (IEC6) was used as an in vitro model, and Confocal laser scanning microscope (CLSM), Flow cytometry (FCM), Indirect immunofluorescence assay (IFA), Western-blotting (WB), and Quantitative real-time RT-PCR (QRT-PCR) methods were used to explore the effects of inflammation on EGF/EGFR biological activity and signal transduction profiles. We found that the EGF/EGFR nuclear signal almost disappeared in the inflammatory state, and the phosphorylation levels of EGFR, AKT, and STAT3 were all significantly down-regulated. In addition, we also studied the effect of ß-carotene on the biological activity of EGF, and found that when cells were pretreated with ß-carotene, the cellular behavior, biological activity, and nuclear signal of EGF/EGFR under inflammation stimulation were partially restored. In summary, the current study shows that inflammation can disrupt EGF/EGFR-mediated signaling in IEC6 cells, suggesting that inflammation negatively regulates the biological activity of EGF/EGFR. Furthermore, we found that ß-carotene not only attenuated lipopolysaccharide (LPS)-induced inflammation but also partially restored the biological activity of EGF in IEC6 cells, laying a solid foundation for studying the biological functions of EGF and ß-carotene.

The basic route of the nuclear translocation porcine growth hormone (GH)-growth hormone receptor (GHR) complex (pGH/GHR) in porcine hepatocytes.

Traditional views suggest that growth hormone and the growth hormone receptor (GH/GHR complex) exert their functions only on the plasma membrane. This paradigm, however, has been challenged by recent new findings that the GH/GHR complex could translocate into cell nuclei where they could still exhibit important physiological functions. We also reported the nuclear localization of porcine GH/GHR and their potential functions in porcine hepatocytes. However, the basic path of pGH/GHR's nuclear translocation remains unclear. Combining previous research results and our current findings, we proposed two basic routes of pGH/GHR's nuclear transportation as follows: 1) after pGH binding to GHR, pGH/GHR enters into the cytoplasm though clathrin- or caveolin-mediated endocytosis, then the pGH/GHR complex enters into early endosomes (Rab5-positive), and the endosome carries the GH/GHR complex to the endoplasmic reticulum (ER). After endosome docking on the ER, the endosome starts fission, and the pGH/GHR complex enters into the ER lumen. Then the pGH/GHR complex transports into the cytoplasm, possibly by the ERAD pathway. Subsequently, the pGH/GHR complex interacts with IMPα/ß, which, in turn, mediates GH/GHR nuclear localization; 2) pGH binds with the GHR on the cell membrane and, subsequently, pGH/GHR internalizes into the cell and enters into the endosome (this endosome may belong to a class of endosomes called envelope-associated endosomes (NAE)). Then, the endosome carries the pGH/GHR to the nuclear membrane. After docking on the nuclear membrane, the pGH/GHR complex fuses with the nuclear membrane and then enters into the cell nucleus.