Hengwei Yu

Low-temperature environments in cold regions pose a significant threat to cattle farming. Bovine mammary epithelial cells (BMECs) are highly sensitive to cold stress, and acute cold stress can induce apoptosis, adversely affecting lactation performance and health. To explore the mechanism of acute cold stress-induced apoptosis in BMECs, we established an in vitro acute cold stress model. Results showed that mRNA levels of HSP90 increased significantly in a time-dependent manner after 2 h of cold stress, confirming successful model establishment. Following 4 h of cold stress, pro-apoptotic genes (Caspase-3, Bax) exhibited significantly elevated mRNA levels, while the anti-apoptotic gene (BCL-2) showed significantly reduced mRNA levels. Concurrently, the apoptosis rate increased significantly, indicating that acute cold stress induces apoptosis and suggesting the 4 h mark may represent a critical transition point. Integrated transcriptomic and functional analyses identified ENO1 as a core metabolic regulator counteracting acute cold stress-induced apoptosis in BMECs. As a multifunctional protein, ENO1 (alpha-enolase) acts as a central enzyme in glycolysis while exerting additional roles in cellular signaling and apoptotic processes, thereby participating in various pathophysiological regulations. Both mRNA and protein levels of ENO1 were significantly elevated in cold-stressed cells compared to untreated controls. Importantly, interference with ENO1 expression aggravated the extent of cold stress-induced apoptosis, demonstrating the regulatory role of ENO1 in this process. To our knowledge, this is the first report elucidating the core regulatory function of ENO1 in acute cold stress-induced apoptosis in BMECs. These findings provide a theoretical basis for understanding apoptotic mechanisms under stress.

Curcumin is a natural polyphenolic compound known to alleviate follicular developmental abnormalities associated with ovarian dysfunction. However, its precise molecular mechanisms remain to be fully elucidated. In this study, we systematically investigated the regulatory effects of curcumin on bovine ovarian granulosa cells through integrated transcriptomic and metabolomic analyses. A total of 503 and 200 significantly altered metabolites were identified in the positive and negative ion modes, respectively, with enrichment in key pathways such as glutathione metabolism, fatty acid biosynthesis, and the phosphatidylinositol signaling pathway. Transcriptomic profiling revealed 1168 differentially expressed genes (582 upregulated and 586 downregulated) which were significantly enriched in pathways related to glutathione metabolism and cellular senescence. Joint multi-omics analysis further demonstrated that curcumin significantly influenced pathways related to glutathione metabolism, cysteine, and methionine metabolism, as well as multiple forms of programmed cell death, including apoptosis, necroptosis, and ferroptosis. Collectively, these findings suggest that curcumin may enhance the antioxidant capacity and survival of granulosa cells by maintaining redox homeostasis and modulating cell fate. This work provides new insights into the potential cellular mechanisms underlying the protective effects of curcumin on granulosa cell function.

FTO expression correlates with adipose tissue development in rabbits, yet its regulatory role in preadipocyte differentiation remains poorly understood. Therefore, we employed transcriptome sequencing to identify 422 differentially expressed genes (DEGs) between the FTO overexpression group and the FTO-negative control group. Multi-omics evidence from proliferation assays (CCK-8), protein verification (WB), gene quantification (qPCR), and lipid staining (Oil Red O) established FTO as a positive regulator of rabbit preadipocyte development with gain-of-function enhancing and loss-of-function impeding differentiation. In conclusion, FTO regulates the proliferation and differentiation of rabbit preadipocytes, providing deeper insights into livestock energy metabolism and physiological functions. These findings hold significant implications for healthy animal farming, high-quality livestock product production, and the prevention and control of metabolic diseases in animals.

Elevated non-esterified fatty acid (NEFA) levels are closely associated with metabolic disorders in dairy cattle, yet their direct effects on adipocyte physiology remain poorly understood. In this study, we demonstrate that high NEFA concentrations significantly impair bovine adipocyte function by simultaneously inhibiting proliferation/differentiation (p < 0.01) and promoting pathological lipid deposition. Through integrated transcriptomic and functional analyses, we identified NDUFAB1 as a central metabolic regulator that counteracts NEFA-induced adipocyte dysfunction. Mechanistically, NDUFAB1 activation attenuates the cytotoxic effects of excessive NEFA exposure. These findings provide both fundamental insights into energy metabolism regulation and a potential therapeutic target (NDUFAB1) for preventing bovine metabolic diseases.

MiR-211-5p exhibits dysregulated expression in various malignant tumors and has been implicated in the regulation of tumor cell proliferation, apoptosis, inflammation, and neurogenic processes. Previous studies have demonstrated that miR-211 negatively regulates ELOVL6, suggesting its involvement in lipid metabolism and lipogenesis within bovine adipose tissue. Our prior transcriptomic analysis revealed upregulated miR-211-5p expression in rabbits fed a high-fat diet, indicating its potential role in lipid metabolism regulation. However, the precise functions of miR-211-5p in lipid deposition and lipogenesis in rabbit preadipocytes remain unclear. To address this knowledge gap, this study utilized rabbit preadipocytes as experimental models to investigate the molecular mechanisms by which miR-211-5p regulates preadipocyte proliferation and differentiation. The findings aim to provide a theoretical basis for improving rabbit meat quality. The main findings of this study are summarized as follows: (1) The EdU proliferation assay, RT-qPCR detection, and CCK-8 cell viability assay revealed that overexpression of miR-211-5p inhibits the proliferation of rabbit preadipocytes, while inhibition of miR-211-5p expression promotes the proliferation of preadipocytes. (2) The precursor adipocytes were transfected and induced to differentiate. RT-qPCR, western blot (WB), and Oil Red O staining assays showed that overexpression of miR-211-5p promotes the maturation and differentiation of precursor adipocytes in meat rabbits, while inhibition of miR-211-5p expression inhibits the maturation and differentiation of precursor adipocytes in rabbits. (3) Through transcriptome sequencing, a total of 147 differentially expressed genes were identified. Among them, TPK1 is the target gene of miR-211-5p and is also the newly identified important gene involved in lipid synthesis. (4) After silencing the target gene TPK1, a series of experiments, including RT-qPCR, WB, Oil Red O staining, and CCK-8 cell viability assay, were conducted. The results showed that interfering with the expression of the TPK1 gene can inhibit the proliferation of rabbit preadipocytes and promote their differentiation. (5) After co-transfection of miR-211-5p inhibitor and si-TPK1, experiments such as EdU assay, RT-qPCR, western blot (WB), Oil Red O staining, and CCK-8 cell viability detection were conducted. It was found that miR-211-5p inhibits the proliferation and promotes the differentiation of rabbit preadipocytes by targeting TPK1.

The growth, development, and milk production traits of dairy goats, which are important sources of high-quality animal protein, are significantly influenced by a combination of genetic and environmental factors. It is imperative to identify key genetic loci that govern economically valuable traits in order to enhance breeding programs. Despite advancements in genomic technologies, there are still gaps in knowledge regarding the interplay between genetic factors and environmental influences, particularly in relation to the regulation of milk production and quality. Therefore, the aim of this paper was to synthesize advancements in the genetic and environmental factors affecting milk production and quality in dairy goats and identify key regulatory mechanisms. This review summarizes the recent progress on the identification of genes associated with milk production traits using whole-genome resequencing, the use of transcriptomic profiling to identify genes linked to milk production, the exploration of regulatory mechanisms of lipid metabolism in goat mammary epithelial cells, and the evaluation of the influence of nutritional factors on milk quality. A comprehensive understanding of these interactions is essential for enhancing breeding strategies and ensuring the sustainability of dairy goat farming. Future research should incorporate multi-omics approaches to unravel the intricate regulatory processes governing milk production and adapt practices to meet global demand while upholding economic and environmental sustainability.

Leptin can indirectly regulate fatty‐acid metabolism and synthesis in muscle in vivo and directly in incubated muscle ex vivo. In addition, non‐synonymous mutations in the bovine leptin gene ( LEP ) are associated with carcass intramuscular fat (IMF) content. However, the effects of LEP on lipid synthesis of adipocytes have not been clearly studied at the cellular level. Therefore, this study focused on bovine primary intramuscular preadipocytes to investigate the effects of LEP on the proliferation and differentiation of intramuscular preadipocytes, as well as its regulatory mechanism in lipid synthesis. The results showed that both the LEP and leptin receptor gene ( LEPR ) were highly expressed in IMF tissues, and their mRNA expression levels were positively correlated at different developmental stages of intramuscular preadipocytes. The overexpression of LEP inhibited the proliferation and differentiation of intramuscular preadipocytes, while interference with LEP had the opposite effect. Additionally, LEP significantly promoted the phosphorylation level of AMPKα by promoting the protein expression of CAMKK2. Meanwhile, rescue experiments showed that the increasing effect of AMPK inhibitors on the number of intramuscular preadipocytes was significantly weakened by the overexpression of LEP . Furthermore, the overexpression of LEP could weaken the promoting effect of AMPK inhibitor on triglyceride content and droplet accumulation, and prevent the upregulation of adipogenic protein expression (SREBF1, FABP4, FASN, and ACCα) caused by AMPK inhibitor. Taken together, LEP acted on the AMPK signaling pathway by regulating the protein expression of CAMKK2, thereby downregulating the expression of proliferation‐related and adipogenic‐related genes and proteins, ultimately reducing intramuscular adipogenesis.

The intramuscular fat (IMF) content of beef determined the meat quality, and the market value of beef varies with different breeds. To provide some new approaches for improving meat quality and cattle breed improvement, 24-month-old Qinchuan cattle (Q, n = 6), Nanyang cattle (N, n = 6), and Japanese black cattle (J, n = 6) were selected. IMF content of the J group (16.92 ± 1.08%) is remarkably higher than that of indigenous Chinese cattle (Q, 13.38 ± 1.08%, and N, 12.35 ± 1.22%). Monounsaturated fatty acids and polyunsaturated fatty acids in the J group are higher than the Q and creatine, lysine, and glutamine are the three most abundant amino acids in beef, which contribute to the flavor formation. Similarly, IMF content-related genes were enriched in four vital KEGG pathways, including fatty acid metabolism, biosynthesis of unsaturated fatty acids, fatty acid elongation, and insulin resistance. Moreover, weighted genes coexpression network analysis (WGCNA) revealed that ITGB1 is the critical gene associated with the IMF content. This study compares transcriptome and metabolome of local and high-IMF cattle breeds, providing data for native cattle breeding and improvement of beef quality.