{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,6]],"date-time":"2026-05-06T06:56:55Z","timestamp":1778050615438,"version":"3.51.4"},"reference-count":50,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,5,1]],"date-time":"2023-05-01T00:00:00Z","timestamp":1682899200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Calmodulin-dependent protein kinase \u03b2 (CaMKK\u03b2) is closely related to Ca<jats:sup>2+<\/jats:sup> concentration. An increase in Ca<jats:sup>2+<\/jats:sup> concentration in the cytoplasm activates CaMKK\u03b2, and activated CaMKK\u03b2 affects the activities of AMPK and mTOR and induces autophagy. A high-concentrate diet leads to Ca<jats:sup>2+<\/jats:sup> disorder in mammary gland tissue.<\/jats:p><\/jats:sec><jats:sec><jats:title>Objectives<\/jats:title><jats:p>Therefore, this study mainly investigated the induction of mammary gland tissue autophagy by a high-concentrate diet and the specific mechanism of lipopolysaccharide (LPS)-induced autophagy in bovine mammary epithelial cells (BMECs).<\/jats:p><\/jats:sec><jats:sec><jats:title>Material and Methods<\/jats:title><jats:p>Twelve mid-lactation Holstein dairy cows were fed with a 40% concentrate diet (LC) and a 60% concentrate diet (HC) for 3 weeks. At the end of the trial, rumen fluid, lacteal vein blood, and mammary gland tissue were collected. The results showed that the HC diet significantly decreased rumen fluid pH, with a pH lower than 5.6 for more than 3 h, indicating successfully induction of subacute rumen acidosis (SARA). The mechanism of LPS-induced autophagy in BMECs was studied in vitro. First, the cells were divided into a Ctrl group and LPS group to study the effects of LPS on the concentration of Ca<jats:sup>2+<\/jats:sup> and autophagy in BMECs. Then, cells were pretreated with an AMPK inhibitor (compound C) or CaMKK\u03b2 inhibitor (STO-609) to investigate whether the CaMKK\u03b2\u2013AMPK signaling pathway is involved in LPS-induced BMEC autophagy.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The HC diet increased the concentration of Ca<jats:sup>2+<\/jats:sup> in mammary gland tissue and pro-inflammatory factors in plasma. The HC diet also significantly increased the expression of CaMKK\u03b2, AMPK, and autophagy-related proteins, resulting in mammary gland tissue injury. In vitro cell experiments showed that LPS increased intracellular Ca<jats:sup>2+<\/jats:sup> concentration and upregulated protein expression of CaMKK\u03b2, AMPK, and autophagy-related proteins. Compound C pretreatment decreased the expression of proteins related to autophagy and inflammation. In addition, STO-609 pretreatment not only reversed LPS-induced BMECs autophagy but also inhibited the protein expression of AMPK, thereby alleviating the inflammatory response in BMECs. These results suggest that inhibition of the Ca<jats:sup>2+<\/jats:sup>\/CaMKK\u03b2\u2013AMPK signaling pathway reduces LPS-induced autophagy, thereby alleviating inflammatory injury of BMECs.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Therefore, SARA may increase the expression of CaMKK\u03b2 by increasing Ca<jats:sup>2+<\/jats:sup> levels and activate autophagy through the AMPK signaling pathway, thereby inducing inflammatory injury in mammary gland tissue of dairy cows.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2023.1186170","type":"journal-article","created":{"date-parts":[[2023,5,1]],"date-time":"2023-05-01T04:31:53Z","timestamp":1682915513000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":9,"title":["A high-concentrate diet induces inflammatory injury via regulating Ca2+\/CaMKK\u03b2-mediated autophagy in mammary gland tissue of dairy 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