OCX-32基因表达载体构建及其对鸡子宫上皮细胞Ca2+浓度和脂质指标的影响

doi:10.16431/j.cnki.1671-7236.2021.07.006

中国畜牧兽医 ›› 2021, Vol. 48 ›› Issue (7): 2324-2332.doi: 10.16431/j.cnki.1671-7236.2021.07.006

OCX-32基因表达载体构建及其对鸡子宫上皮细胞Ca²⁺浓度和脂质指标的影响

廖朝美¹, 吴磊¹, 游敏芳¹, 谭光辉¹, 李杰章¹, 李万贵², 张依裕¹

1. 贵州大学动物科学学院, 高原山地动物遗传育种与繁殖教育部重点实验室, 贵阳 550025;
2. 六盘水市草地工作站, 六盘水 553000

修回日期:2020-11-02 出版日期:2021-07-20 发布日期:2021-07-15
通讯作者: 张依裕 E-mail:zyy8yyc@163.com
作者简介:廖朝美(1997-),女,贵州黄平人,硕士生,研究方向:动物遗传育种与繁殖,E-mail:3285438093@qq.com
基金资助:
贵州省科技合作计划项目（黔科合LH字〔2016〕7454号）

Construction of OCX-32 Gene Expression Vector and Its Effect on Ca²⁺ Concentration Lipid Indexes of Chicken Uterine Epithelial Cells

LIAO Chaomei¹, WU Lei¹, YOU Minfang¹, TAN Guanghui¹, LI Jiezhang¹, LI Wangui², ZHANG Yiyu¹

1. Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education, College of Animal Science, Guizhou University, Guiyang 550025, China;
2. Liupanshui City Grassland Workstation, Liupanshui 553000, China

Revised:2020-11-02 Online:2021-07-20 Published:2021-07-15

摘要/Abstract

摘要： 为探究卵黄蛋白原-32（ovocalyxin-32，OCX-32）基因过表达与鸡子宫上皮细胞Ca²⁺含量和脂质指标之间的关联性，本研究采用Ⅳ型胶原酶法分离培养长顺绿壳蛋鸡子宫上皮细胞，构建pcDNA3.1（+）+OCX-32+Flag表达载体并转染子宫上皮细胞，测定OCX-32基因过表达量、Ca²⁺浓度和脂质指标并进行相关性分析。结果显示，试验组的总胆固醇（TCHO）浓度显著高于对照组（P<0.05），对照组的Ca²⁺浓度显著高于试验组（P<0.05）。OCX-32基因过表达量与Ca²⁺浓度呈显著负相关关系（P<0.05），与甘油三酯（TG）浓度呈显著正相关关系（P<0.05），与TCHO浓度呈极显著负相关关系（P<0.01），TCHO浓度与TG浓度呈极显著负相关关系（P<0.01），研究结果揭示了OCX-32基因过表达能够调控鸡子宫上皮细胞的Ca²⁺含量和脂质的合成转运，为深入阐释鸡子宫上皮细胞中脂质和钙分泌的调控机制提供试验基础和数据支持。

关键词: OCX-32基因; 长顺绿壳蛋鸡; 子宫上皮细胞; Ca²⁺; 脂质指标

Abstract: In order to explore the relationship between ovocalyxin-32 (OCX-32) gene overexpression and Ca²⁺ concentration and lipid indexes of chicken uterine epithelial cells.The Changshun Green-eggshell chicken uterine epithelial cells were isolated and cultured by collagenase Ⅳ method.The expression vector pcDNA3.1(+)+OCX-32+Flag was constructed by homologous recombination method and transfected into uterine epithelial cells.The overexpression of OCX-32 gene,the concentration of Ca²⁺ and the indexes of lipid were determined and the correlation was analyzed.The results showed that the concentration of total cholesterol (TCHO) in the experimental group was significantly higher than that in the control group (P<0.05),and the concentration of Ca²⁺ in the control group was significantly higher than that in the experimental group (P<0.05).The OCX-32 gene overexpression was negatively correlated with Ca²⁺ concentration (P<0.05),and significant positive correlation with triglyceride (TG) concentration (P<0.05),and extremely significant negative correlated with TCHO concentration (P<0.01).Meanwhile,TCHO concentration was extremely significant negative correlated with TG concentration (P<0.01).The results revealed that overexpression of OCX-32 gene could regulate Ca²⁺ concentration and lipid synthesis and transport in chicken uterine epithelial cells.The results of this study provided experimental basis and data support for further elucidating the regulatory mechanism of lipid and calcium secretion in chicken uterine epithelial cells.

Key words: OCX-32 gene; Changshun Green-eggshell chicken; uterine epithelial cells; Ca²⁺; lipid indexes

中图分类号:

S831.2

廖朝美, 吴磊, 游敏芳, 谭光辉, 李杰章, 李万贵, 张依裕. OCX-32基因表达载体构建及其对鸡子宫上皮细胞Ca²⁺浓度和脂质指标的影响[J]. 中国畜牧兽医, 2021, 48(7): 2324-2332.

LIAO Chaomei, WU Lei, YOU Minfang, TAN Guanghui, LI Jiezhang, LI Wangui, ZHANG Yiyu. Construction of OCX-32 Gene Expression Vector and Its Effect on Ca²⁺ Concentration Lipid Indexes of Chicken Uterine Epithelial Cells[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2324-2332.

参考文献

[1] ZHU M K,LI H Y,MIAO L P,et al.Dietary cadmium chloride impairs shell biomineralization by disrupting the metabolism of the eggshell gland in laying hens[J].Journal of Animal Science,2020,8(2):2-9.
[2] ARIAS J L,NAKAMURA O,FERNANDEZ M S,et al.Role of type collagen on experimental mineralization of eggshell membranes[J].Connective Tissue Research,1997,36(1):21-33.
[3] NYS Y,GAUTRON J,GARCIA R J M.Avian eggshell mineralization:biochemical and functional characterization of matrix proteins[J].Comptes Rendus-Palevol,2004,3(6-7):549-562.
[4] WANG S,HAN J,XIA J,et al.Overexpression of nicotinamide mononucleotide adenylyltransferase (nmnat) increases the growth rate,Ca²⁺ concentration and cellulase production in ganoderma lucidum[J].Applied Microbiology and Biotechnology,2020,104(6936):6-9.
[5] WANG J,YUE H G,WU S,et al.Nutritional modulation of health,egg quality and environmental pollution of the layers[J].Animal Nutrition,2017,3(2):3-8.
[6] PULLI I,ASGHAR M Y,KEMPPAINEN K,et al.Sphingolipid-mediated calcium signaling and its pathological effects[J].Biochimica et Biophysica Acta (BBA)-Molecular Cell Research,2018,10(4):32-36.
[7] MIRANDA R G A,ARAUJO-CHAVES J C,KAWAI C,et al.Cardiolipin structure and oxidation are affected by Ca²⁺ at the interface of lipid bilayers[J].Frontiers in Chemistry,2020,7(23):925-930.
[8] FLEISCHMAN A I,YACOWITZ H,HAYTON T,et al.Effects of dietary calcium upon lipid metabolism in mature male rats fed beef tallow[J].Journal of Nutrition,1966,88(3):255-260.
[9] 徐姝迪.高钙饲料喂养初断乳大鼠对肥胖诱导形成的影响及其机制研究[D].乌鲁木齐:新疆医科大学,2011. XU Z D.Study on influence and mechanism of ablactation rats' high calcium intake to adulthood obesity[D].Urumqi:Xinjiang Medical University,2011.(in Chinese)
[10] RODAS B Z D,GILLILAND S E,MAXWELL C V.Hypocholesterolemic action of Lactobacillus acidophilus ATCC 43121 and calcium in swine with hyperchole-sterolemia induced by diet[J].Journal of Dairy Science,1996,79(12):2121-2128.
[11] 王丽.钙对小鼠脂质蓄积和分解的调节作用及其机制研究[D].杨凌:西北农林科技大学,2009. WANG L.Effect and mechanism of calcium on lipid accumulation and lipoclaclasis in mice[D].Yangling:Northwest A&F University,2009.(in Chinese)
[12] 罗华伦.鸭SCD-1基因对子宫上皮细胞Ca²⁺的效应及其启动子活性研究[D].贵阳:贵州大学,2019. LUO H L.Effect of duck SCD-1 gene on Ca²⁺ in uterine epithelial cells and its promoter activity research[D].Guiyang:Guizhou University,2019.(in Chinese)
[13] LIANG S,CHEN J,NIU J,et al.Study on utilization of eggshell and scallop shell in microbial induced calcium carbonate precipitation[J].Environmental Science & Technology,2019,6(9):83-86.
[14] SOCHA J K,HRABIA A.Alterations in apoptotic markers and egg-specific protein gene expression in the chicken oviduct during pause in laying induced by tamoxifen[J].Theriogenology,2018,105(8):126-134.
[15] LAZZINI S.Novel SNPs in the OCX-32 gene and their associations with egg production traits in Korean native chickens[J].Plant & Animal Genome,2015,44(1):16-19.
[16] 张亚男.饲粮锌对产蛋后期蛋鸡蛋壳品质及抗氧化机能的影响[D]:北京:中国农业科学院,2013. ZHANG Y N.Effects of dietary zinc on eggshell quality and antioxidant status of old laying hens[D].Beijing:Chinese Academy of Agricultural Sciences,2013.(in Chinese)
[17] VASKONEN T,MERVAALA E,SUMUVUORI V,et al.Effects of calcium and plant sterols on serum lipids in obese Zucker rats on a low-fat diet[J].British Journal of Nutrition,2002,87(3):239-245.
[18] BALLA T.Regulation of Ca²⁺ entry by inositol lipids in mammalian cells by multiple mechanisms[J].Cell Calcium,2009,45(6):527-534.
[19] 刘德文.血浆低密度脂蛋白(LDL)及高密度脂蛋白(HDL)的来源[J].山西医学院学报,1985,26(3):31-35. LIU D W.Sources of plasma low-density lipoprotein (LDL) and high-density lipoprotein (HDL)[J].Journal of Shanxi Medical University,1985,26(3):31-35.(in Chinese)
[20] MA K Y,YANG N,JIAO R,et al.Dietary calcium decreases plasma cholesterol by down-regulation of intestinal Niemann-Pick C1 like 1 and microsomal triacylglycerol transport protein and up-regulation of CYP7A1 and ABCG 5/8 in hamsters[J].Molecular Nutrition & Food Research,2011,55(2):247-258.
[21] OJUKA E O.Role of calcium and AMP kinase in the regulation of mitochondrial biogenesis and GLUT4 levels in muscle[J].Proceedings of the Nutrition Society,2004,63(2):275-283.
[22] WRIGHT D C,HUCKER K A,HOLLOSZY J O,et al.Ca²⁺ and AMPK both mediate stimulation of glucose transport by muscle contractions[J].Diabetes,2004,53(2):330-335.
[23] WOLIDE A D,ZAWDIE B,ALEMAYEHU T,et al.Association of trace metal elements with lipid profiles in type 2 diabetes mellitus patients:A cross sectional study[J].BMC Endocrine Disorders,2017,17(1):64-68.
[24] 肖俊峰,武书庚,张海军,等.四种壳基质蛋白研究进展[J].中国家禽,2012,34(9):44-47. XIAO J F,WU S G,ZHANG H J,et al.Research progress of four shell matrix proteins[J].China Poultry,2012,34(9):44-47.(in Chinese)
[25] ZHANG L P,ZENG T,LU L Z,et al.Correlation analysis of single nucleotide polymorphisms of cellular retionol-binding protein 4(CRBP4) and ovocalyxin-32(OCX-32) genes with egg production traits in Lingkun chicken (Gallus gallus)[J].Journal of Agricultural Biotechnology,2015,276(42):39-52.
[26] BARAN D T,QUAIL J M,RAY R,et al.Annexin Ⅱ is the membrane receptor that mediates the rapid actions of 1alpha,25-dihydroxyvitamin D(3)[J].Journal of Cellular Biochemistry,2000,78(1):34-46.
[27] 李杰章,罗华伦,谭光辉,等.SCD-1基因过表达对鸭子宫上皮细胞钙离子和脂质含量的效应[J].生物工程学报,2020,36(5):899-907. LI J Z,LUO H L,TAN G H,et al.Effects of SCD-1 gene overexpression on the content of calcium ion and lipids in duck uterine epithelial cells[J].Chinese Journal of Biotechnology,2020,36(5):899-907.(in Chinese)

OCX-32基因表达载体构建及其对鸡子宫上皮细胞Ca²⁺浓度和脂质指标的影响

Construction of OCX-32 Gene Expression Vector and Its Effect on Ca²⁺ Concentration Lipid Indexes of Chicken Uterine Epithelial Cells

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