南方农业学报2024，Vol.55Issue(7)：1887-1896,10.DOI:10.3969/j.issn.2095-1191.2024.07.002
相思子碱通过P53/mTOR通路缓解脂多糖对水牛乳腺上皮细胞β-酪蛋白合成的影响
Abrine mitigates the effects of lipopolysaccharide on β-casein synthesis in buffalo mammary epithelial cells by modulating the P53/mTOR pathway
摘要
Abstract
[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.关键词
水牛乳腺上皮细胞/相思子碱/脂多糖/β-酪蛋白/P53/mTOR通路Key words
buffalo mammary epithelial cells/abrine/lipopolysaccharide/β-casein/P53/mTOR pathway分类
农业科技引用本文复制引用
周金陈,尚江华,肖鹏,李孟琪,张博,郑海英,杨春艳,田新如,黄依林,杨小淦..相思子碱通过P53/mTOR通路缓解脂多糖对水牛乳腺上皮细胞β-酪蛋白合成的影响[J].南方农业学报,2024,55(7):1887-1896,10.基金项目
国家自然科学基金项目(31872350) （31872350）
广西科技重大专项(桂科AA22068099) （桂科AA22068099）
广西自然科学基金项目(2021GXNSFAA 196021) （2021GXNSFAA 196021）
国家现代农业产业技术体系广西奶水牛产业创新团队项目(nycytxgxcxtd-2021-21-01) National Natural Science Foundation of China(31872350) （nycytxgxcxtd-2021-21-01）
Guangxi Science and Technology Major Project(Guike AA22068099) （Guike AA22068099）
Guangxi Natural Science Foundation(2021GXNSFAA196021) （2021GXNSFAA196021）
Guangxi Dairy Bu-ffalo Industry Innovation Team Project of China Agriculture Research System(nycytxgxcxtd-2021-21-01) （nycytxgxcxtd-2021-21-01）
