Abrine mitigates the effects of lipopolysaccharide on β-casein synthesis in buffalo mammary epithelial cells by modulating the P53/mTOR pathway
[Objective]The objective of this study was to investigate the mitigating effect of abrine on induced inhibi-tion by lipopolysaccharide(LPS)of casein synthesis in buffalo mammary epithelial cells by modulating the P53/mTOR pathway,to provide a theoretical basis for mitigating the decline in milk quality in buffaloes caused by mastitis.[Method]The LPS-induced inflammation model was established in buffalo mammary epithelial cells,co-incubated with abrine,P53 inhibitor(Pifithrin-μ),P53 activator(Kevetrin hydrochloride)and buffab mammary epithelial cells for 12 h;morpho-logical observations of mammary tissues were conducted using HE staining;cell viability was assessed using the CCK-8 method,while the secretion levels of β-casein were determined using ELISA kits;real-time quantitative PCR was em-ployed to measure the gene relative expression levels of NF-κB,IL-1β,TNFα,β-casein,mTOR,P53,JAK2,STAT5,and AKT1;additionally,Western blotting was used to assess the protein relative expression levels of NF-κB,β-casein,mTOR,and P53.[Result]Clinical mastitis(CMS)mammary tissues showed infiltration of neutrophils in the interstitium and alveolar cavities,accompanied by atrophy at certain degree in some alveolar structures.The buffalo mammary epithe-lial cell viability of the 1.0 μg/mL LPS treatment group was significant decreased(P<0.05,the same below),buffab mam-mary epithelial cell viability of 2.0,4.0,and 8.0 μg/mL LPS treatments extremely significantly(P<0.001).Abrine at 25,50,100,and 200 μmol/L alleviated the impact of LPS,resulting in obvious increase in cell viability.Compared to the blank control group,LPS treatment group significantly elevated relative expression NF-κB,IL-1β,TNFα and P53 genes and proteins in buffalo mammary epithlial cells,while extremely significantly reduced β-casein expression.Abrine co-treatment with LPS group resulted in significant increase in β-casein levels in buffalo mammary epithlial cells and signifi-cant decrease in NF-κB,IL-1β,TNFα and P53 genes and proteins relative expression compared to the LPS group.The LPS+Pifithrin-μ group showed significant increase in β-casein expression in buffalo mammary epithlial cells compared to the LPS group.Conversely,abrine+Kevetrin hydrochloride group exhibited significant decrease in β-casein expression in buffalo mammary epithlial cells compared to abrine group.[Conclusion]LPS decreases β-casein synthesis and expression levels of genes and their encoded proteins involved in β-casein synthesis;abrine alleviates LPS-induced mammary epithe-lial cells of buffalo by inhibiting NF-κB pathway,and alleviates LPS-induced reduction of β-casein synthesis in buffalo mammary epithelial cells by modulating P53/mTOR pathway.
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