奶水牛ACSL1基因组织表达分析及其对脂肪酸代谢相关基因表达的影响  

Expression of ACSL1 and Its Effect on Expression Involved in Fatty Acid Metabolism in Buffalo

作　　者：梁莎莎 庞春英[1] 邓廷贤[1] 马小娅 陆杏蓉 段安琴 梁贤威[1] LIANG Shasha;PANG Chunying;DENG Tingxian;MA Xiaoya;LU Xingrong;DUAN Anqin;LIANG Xianwei(Guangxi Key Laboratory of Buffalo Genetics,Breeding and Reproduction,Guangxi Buffalo Research Institute,Chinese Academy of Agricultural Sciences,Guangxi Nanning 530001,China)

机构地区：[1]中国农业科学院广西水牛研究所,农业部(广西)水牛遗传繁育重点实验室,广西南宁530001

出　　处：《中国畜牧杂志》2020年第5期41-45,共5页Chinese Journal of Animal Science

基　　金：广西科技重大专项(桂科AA16450002);广西自然科学基金(2017GXNSFAA198108、2017GXNSFBA198191、2017GXNSFBA198022);广西水牛研究所基本科研业务费(180502);广西水牛研究所基本科研业务费(201905007);广西水牛研究所基本科研业务费(180501);2019年广西农业科技自筹经费项目(Z201948)。

摘　　要：本实验通过开展奶水牛长链脂酰辅酶A合成酶1(Long-chain fatty acyl-CoA 1,ACSL1)基因的编码区克隆、组织表达分析、过表达及干扰等实验,旨在探究该基因在乳腺上皮细胞中对脂肪酸代谢相关基因表达水平的影响。组织表达分析结果显示:ACSL1在水牛乳腺组织中的表达量最高,暗示该基因可能与泌乳期水牛乳脂合成有关。鉴于此,本实验成功克隆了水牛ACSL1基因完整的编码序列,由此构建了相应的过表达载体,并合成了干扰片段。水牛乳腺上皮细胞ACSL1超表达和干扰实验发现:与对照组(感染AdpcDNA3.1+)相比,实验组(感染Ad-ACSL1)CLIC1、ELOVL1和PNPLA2显著上升,ELOVL6表达量极显著上升,ACSL1、CPT1B、CPT1C和DDR1表达量极显著上升,FADS2差异不显著;与对照组(感染siRNA-NC)相比,实验组(感染siRNA-ACSL1)ELOVL1和FADS2表达量显著下降,ACSL1、CLIC1、CPT1C、ELOVL6、DDR1和PNPLA2表达量极显著下降,CPT1B差异不显著。这些结果说明ACSL1基因的过表达或干扰可能影响水牛乳腺上皮细胞中部分脂代谢相关基因的表达量变化,从而影响乳脂合成。This study was conducted to study the effect of Long-chain fatty acyl-CoA 1(ACSL1)gene on the expression level of fatty acid metabolism related genes in mammary epithetical cells after its overexpression and interference,tissue expression analysis,overexpression and interference technology.The Real-time PCR results showed that the expression of buffalo ACSL1 gene in the mammary gland was obviously higher than other tissues,It is suggested that this gene may be involved in lactating buffalo milk fat synthesis.RNA was extracted from buffalo mammary epithelial cells transfected with overexpression vectors and siRNA 48 h later.qPCR results showed that compared with the control group(Ad-pcDNA3.1),the experimental group(Ad-ACSL1)showed a significant increase in CLIC1,ELOVL1 and PNPLA2(P<0.05),the expression of ELOVL6(P<0.01),the expression of ACSL1,CPT1 B,CPT1 C and DDR1(P<0.001),and the difference of FADS2 was not significant(P>0.05).Compared with the control group(siRNA-NC),the expression of ELOVL1 and FADS2 in the experimental group(siRNA-ACSL1)decreased significantly(P<0.05),the expression of ACSL1,CLIC1,CPT1 C,ELOVL6,DDR1 and PNPLA2 decreased significantly(P<0.01),and there was no significant difference of CPT1 B(P>0.05).Therefore,the overexpression or interference of ACSL1 gene may affect the expression of some lipid metabolism-related genes in buffalo mammary epithetical cells,thus affecting milk fat synthesis.

关 键 词：水牛 长链脂酰辅酶A合成酶1 组织表达 过表达 干扰 

分 类 号：S823.2[农业科学—畜牧学]