氯化两面针碱通过Wnt/mTOR信号通路缓解热应激水牛乳腺上皮细胞β-酪蛋白含量降低
Nitidine chloride alleviates the decrease of β-casein synthesis content in buffalo mammary epithelial cells caused by heat stress through Wnt/mTOR signaling pathway
张博 1肖鹏 1李孟琪 1周金陈 1尚江华 2郑海英 2田新如 1黄依林 1杨小淦 3杨春艳2
作者信息
- 1. 广西大学动物科学技术学院,广西南宁 530004;广西水牛研究所/广西水牛遗传繁育重点实验室,广西南宁 530001
- 2. 广西水牛研究所/广西水牛遗传繁育重点实验室,广西南宁 530001
- 3. 广西大学动物科学技术学院,广西南宁 530004
- 折叠
摘要
[目的]探究氯化两面针碱(Nitidine chloride,NitC)对热应激水牛乳腺上皮细胞β-酪蛋白含量降低的影响,为实际生产中缓解热应激对水牛乳品质的负面影响提供理论依据.[方法]记录非热应激季节和热应激季节牛舍温湿度指数(Temperature-humidity index,THI),检测水牛呼吸频率和直肠温度以判断水牛是否处于热应激状态;采集奶样进行乳成分分析以探究热应激对水牛奶β-酪蛋白含量的影响;以水牛乳腺上皮细胞为研究对象,经40.5℃处理24 h建立细胞热应激模型,使用LiCl和LGK974分别激活和抑制Wnt通路,检测水牛乳腺上皮细胞β-酪蛋白含量及β-酪蛋白合成相关基因和蛋白的表达水平,探究氯化两面针碱缓解热应激水牛乳腺上皮细胞β-酪蛋白含量降低的作用机制.[结果]与非热应激季节相比,热应激季节牛舍THI升高并超过水牛热应激阈值,水牛呼吸频率和直肠温度显著升高(P<0.05,下同),水牛奶β-酪蛋白含量显著降低;细胞试验结果表明,40.5℃处理24 h能显著降低水牛乳腺上皮细胞β-酪蛋白含量、细胞活力及Wnt2、mTOR、β-casein基因和蛋白的相对表达量;而氯化两面针碱处理能有效缓解热应激对水牛乳腺上皮细胞的影响,显著提高水牛乳腺上皮细胞β-酪蛋白含量、细胞活力及Wnt2、mTOR、β-casein基因和蛋白的相对表达量.[结论]热应激降低了水牛乳腺上皮细胞β-酪蛋白含量及β-酪蛋白合成相关基因和蛋白的相对表达量,氯化两面针碱则通过Wnt/mTOR信号通路提高水牛乳腺上皮细胞β-酪蛋白含量、细胞活力及Wnt2、mTOR、β-casein基因和蛋白的相对表达量,以缓解热应激造成的水牛乳腺上皮细胞β-酪蛋白含量降低.
Abstract
[Objective]This study aimed to explore the effects of nitidine chloride(NitC)on the reduction of β-casein content in buffalo mammary epithelial cells caused by heat stress,in order to provide a theoretical basis for alleviating the negative impact of heat stress on buffalo milk quality in the actual production.[Method]The temperature-humidity index(THI)of the cowshed was recorded in the non-heat stress season and the heat stress season,and the respiratory rate and rectal temperature of the buffalo were detected to determine whether the buffalo was in a state of heat stress;milk samples were collected for milk composition analysis to investigate the effects of heat stress on the β-casein content in buffalo milk;bu-ffalo mammary epithelial cells were used as the research objects,and the buffalo mammary epithelial cells heat stress model was established by treating them at 40.5℃for 24 h.LiCl and LGK974 were used to activate and inhibit the Wnt pathway respectively,and the β-casein content in buffalo mammary epithelial cells and the expression levels of genes and proteins related to β-casein synthesis were detected to explore the mechanism by which nitidine chloride alleviated the decrease of β-casein content in buffalo mammary epithelial cells caused by heat stress.[Result]Compared with the non-heat stress season,the THI in the cowshed increased and exceeded the heat stress threshold of buffaloes in the heat stress season,the respiratory rate and rectal temperature of buffaloes increased significantly(P<0.05,the same below),and the β-casein content in buffalo milk decreased significantly.Cell experiments showed that treatment with 40.5℃for 24 h significantly reduced the β-casein content,cell viability,and the relative expression of Wnt2,mTOR and β-casein genes and proteins in buffalo mammary epithelial cells.Nitidine chloride treatment effectively alleviated the effects of heat stress on buffalo mammary epithelial cells and significantly increased the β-casein content,cell viability and the relative expression of Wnt2,mTOR and β-casein genes and proteins in buffalo mammary epithelial cells.[Conclusion]Heat stress reduces the β-casein content and the relative expression of β-casein synthesis-related genes and proteins in buffalo mammary epithelial cells.Nitidine chloride increases the β-casein content,cell viability and the relative expression of Wnt2,mTOR and β-casein genes and proteins in buffalo mammary epithelial cells through the Wnt/mTOR signaling pathway,alleviating the decrease in β-casein content in buffalo mammary epithelial cells caused by heat stress.
关键词
氯化两面针碱/热应激/水牛乳腺上皮细胞/Wnt/mTOR通路/β-酪蛋白Key words
nitidine chloride/heat stress/buffalo mammary epithelial cells/Wnt/mTOR pathway/β-casein引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据