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| SCFAs对奶牛瘤胃上皮细胞Ca2+信号通路相关基因表达的影响 |
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宁丽丽1,成健4,詹康1,杨天宇1,姜茂成1,赵国琦1,2,3*
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| (1.扬州大学 动物科学与技术学院, 江苏 扬州 225009;2.扬州大学 农业科技发展研究院, 江苏 扬州 225009;3.扬州大学 教育部农业与农产品安全国际合作联合实验室, 江苏 扬州 225009;4.常熟市海虞动物防疫站, 江苏 常熟 215500) |
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| 摘要: |
| 为探究短链脂肪酸(SCFAs)对奶牛瘤胃上皮细胞(BRECs)Ca2+信号通路相关基因表达的影响，试验分为3个处理组，分别为野生型BRECs组，含20 mmol/L SCFAs BRECs组，含20 mmol/L SCFAs且通过CRISPR/Cas 9系统敲除GPR41基因的BRECs组，每个处理组3个重复，每组细胞均培养24 h后，收集细胞提取总RNA，通过qRT-PCR对Ca2+信号通路相关基因的mRNA表达量和细胞内Ca2+浓度进行测定。结果表明，与野生型BRECs相比，添加20 mmol/L SCFAs可极显著增加PLCB2的mRNA表达量(P<0.01)，显著增加IP3R1的mRNA表达量(P<0.05)，对PLCE1、PLCL1、PKCB和PKCG的mRNA表达量无显著差异(P>0.05)，可增加细胞内Ca2+浓度但无显著差异(P>0.05)；敲除SCFAs的受体GPR41后，添加20 mmol/L SCFAs可显著降低IP3R1的mRNA表达量(P<0.05)，极显著上调PLCE1和PLCB2的mRNA表达量(P<0.01)，但对PLCL1、PKCB和PKCG的mRNA表达量无显著影响(P>0.05)，细胞内Ca2+浓度有降低趋势但无显著差异(P>0.05)。综上，SCFAs可以通过激活其受体GPR41来调控BRECs内Ca2+信号通路中相关基因的表达和细胞内Ca2+的释放。 |
| 关键词: 瘤胃上皮细胞 短链脂肪酸 G蛋白偶联受体 钙离子 |
| DOI：10.11841/j.issn.1007-4333.2020.08.06 |
| 投稿时间：2019-12-04 |
| 基金项目:现代农业产业技术体系专项资金项目(CARS-36)；国家自然科学基金项目(31972589) |
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| Effects of SCFAs on the mRNA expressions of related genes in Ca2+ signaling pathways in the rumen epithelial cells of dairy cow |
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NING Lili1,CHENG Jian4,ZHAN Kang1,YANG Tianyu1,JIANG Maocheng1,ZHAO Guoqi1,2,3*
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| (1.College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;2.Institutes of Agricultural Science and Technology Development, Yangzhou University, Yangzhou 225009, China;3.China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education, Yangzhou University, Yangzhou 225009, China;4.Changshu Haiyu Animal Epidemie Prevention Station, Changshu 215500, China) |
| Abstract: |
| To investigate the effects of SCFAs on the mRNA expressions of related genes in Ca2+ signaling pathways in the rumen epithelial cells of dairy cow, three groups were divided, which were wild BRECs group, wild BRECs with 20 mM SCFAs group and GPR41 KO BRECs with 20 mM SCFAs group, respectively. After culture of 24 h, the total RNA was extracted from the collected cells. qRT-PCR was employed for analysis of mRNA expression and intracellular Ca2+ concentration of the Ca2+ signaling pathway-related genes. The results showed that PLCB2(P<0. 01)and IP3R1(P<0. 05)were significantly up-regulated after the addition of 20 mM SCFAs in the wild-type BRECs. SCFAs also increased the concentration of intracellular Ca2+ but had no significant difference(P>0. 05). There was no significant difference in the mRNA expression levels of PLCE1, PLCL1, PKCB and PKCG(P>0. 05). Additionally, after the GPR41 was knocked out, the mRNA expression of IP3R1 was significantly down-regulated(P<0. 05). The mRNA expression levels of PLCE1 and PLCB2 were significantly upregulated(P<0. 01). Intracellular Ca2+ concentration showed a decreasing trend but showed no significant difference(P>0. 05). There was no significant effect on PLCL1, PKCB and PKCG(P>0. 05). In conclusion, SCFAs could regulate the expressions of related genes and the release of Ca2+ in the BRECs by activating its receptor GPR41. |
| Key words: rumen epithelial cells short chain fatty acids G protein-coupled receptor calcium ion |
