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        The RCAN1.4-calcineurin/NFAT signaling pathway is essential for hypoxic adaption of intervertebral discs

        Huang Bao,He Yongqing,Li Shengwen,Wei Xiaoan,Liu Junhui,Shan Zhi,Huang Yue,Chen Jiang,Zhao Fengdong 생화학분자생물학회 2020 Experimental and molecular medicine Vol.52 No.-

        Calcipressin-1, also known as regulator of calcineurin 1 (RCAN1), can specifically bind calcineurin at or near the calcineurin A catalytic domain and downregulate calcineurin activity. However, whether RCAN1 affects the hypoxic intervertebral disc (IVD) phenotype through the calcineurin/NFAT signaling pathway remains unclear. First, we confirmed the characteristics of the degenerative nucleus pulposus (NP) by H&E, safranin O/fast green and Alcian blue staining, and detected increased RCAN1 levels in the degenerative NP by immunohistochemistry. Then, we demonstrated that the protein level of RCAN1.4 was higher than that of RCAN1.1 and progressively elevated from the control group to the Pfirrmann grade V group. In vitro, both hypoxia (1% O2) and overexpression of HIF-1α reduced the protein level of RCAN1.4 in rat NP cells in a dose- and time-dependent manner. We further found that miRNA-124, through a nondegradative pathway (without the proteasome or lysosome), suppressed the expression of RCAN1.4. As expected, calcineurin in NP cells was activated and primarily promoted nuclear translocation of NFATc1 under hypoxia or RCAN1.4 siRNA transfection. Furthermore, SOX9, type II collagen and MMP13 were elevated under hypoxia, RCAN1.4 siRNA transfection or NFATc1 overexpression. Using chromatin immunoprecipitation (ChIP) and a luciferase reporter assay (with mutation), we clarified that NFATc1 increasingly bound the SOX9 promotor region (bp −367~−357). Interaction of HIF-1α and NFATc1 promoted MMP13 transcription. Finally, we found that FK506 reversed hypoxia-induced activation of the calcineurin/NFAT signaling pathway in NP cells and an ex vivo model. Together, these findings show that the RCAN1.4-calcineurin/ NFAT signaling pathway has a vital role in the hypoxic phenotype of NP cells. RCAN1.4 might be a therapeutic target for degenerative disc diseases.

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        Indoor feeding combined with restricted grazing time improves body health, slaughter performance, and meat quality in Huang-huai sheep

        Huang Yafeng,Zhao Mengyu,Zhang Xiaoan,Wei Huiqing,Liu Lumeng,Zhang Zi Jun,Cheng Xiao,Wang Guanjun,Ren Chunhuan 아세아·태평양축산학회 2023 Animal Bioscience Vol.36 No.11

        Objective: The aim of this study was to evaluate the effects of three feeding systems, i.e., indoor feeding (CON), indoor feeding with 4-h daily access to grazing artificial pasture (ITGP), and indoor feeding with 8-h daily access to grazing artificial pasture (IEGP), on the plasma antioxidant and immunological capacity, slaughter characteristics, meat quality and economic efficiency of Huang-huai lambs. Methods: Thirty-three healthy Huang-huai rams with similar body weight (approximately 5 mo of age, 28.96±1.01 kg) were assigned equally to three experimental groups. When finished fattening, six lambs from each group were collect blood samples for plasma analyses and then slaughtered to determine slaughter characteristics and obtain biceps brachii muscle for further analysis of meat quality and fatty acid profile. Results: Compared to CON group, animals submitted to ITGP and IEGP groups resulted in greater contents of serum glutathione peroxidase, immunoglobulins (IgA, IgG, and IgM), polyunsaturated fatty acids (PUFA), n-6 PUFA, and PUFA/saturated fatty acid (FA) ratio and lower palmitic /oleic acid ratio (p<0.05). Moreover, animals in ITGP group exhibited a higher (p<0.05) loin eye area, content of meat crude protein (CP), and eicosetrienoic acid compared to CON group, while slaughter performance was superior (p<0.05) to that of the IEGP group. The economic efficiency of ITGP group was 70.12% higher than that of CON group, while the IEGP group exhibited a decrease of 92.54% in economic efficiency compared to the CON group. Conclusion: Restricted grazing time combined with indoor feeding was more effective in conferring superior body health, carcass traits and economic efficiency in Huang-huai lambs, as well as higher CP content and healthier FA composition in the resulting meat.

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