α-ketoglutarate acid attenuates LPS induced necroptosis in bovine ovarian granulosa cells by suppressing autophagy.

The follicle was regarded as the fundamental functional unit of the ovary, and dysfunction of its granulosa cells was known to impair follicular development and delay ovulation. In bovine, reproductive‑tract infections frequently elicited excessive inflammatory responses that resulted in follicular dysfunction, a process largely mediated by lipopolysaccharide (LPS) induced programmed death of ovarian granulosa cells. Necroptosis was shown to contribute to the regulation of follicular development while simultaneously promoting the release of pro‑inflammatory mediators. Although α‑ketoglutarate acid (α‑KG) was recognized as an anti‑inflammatory and antioxidant metabolite, its capacity to attenuate LPS‑induced necroptosis in bovine granulosa cells-and the underlying. To investigate the protective mechanism of α-KG against LPS-induced functional impairment in ovarian granulosa cells, primary granulosa cells were isolated from bovine follicles and cultured in vitro. A series of assays-including qRT-PCR, Western blotting, flow cytometry, immunofluorescence staining, and transmission electron microscopy-were conducted to assess necroptosis, autophagy, and mitochondrial function. LPS was found to induce necroptosis in bovine ovarian granulosa cells by promoting autophagy. This process was accompanied by mitochondrial dysfunction and disruption of mitochondrial cristae structure. Notably, α-KG was shown to inhibit autophagy-mediated necroptosis by restoring mitochondrial membrane potential, decreasing intracellular levels of ROS, Ca²⁺, and thereby attenuating inflammatory responses. In summary, the findings indicated that α-KG attenuated mitochondrial dysfunction-associated autophagy, necroptosis, and inflammatory responses in bovine ovarian granulosa cells following LPS exposure in vitro. These results suggested that α-KG may potentially provide a pharmacological basis for mitigating inflammation-associated follicular dysplasia in bovine, although further in vivo studies are required to confirm these effects.