磷脂磷酸酶3基因与鹅肥肝形成关系的研究
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摘要
为探究磷脂磷酸酶3 (PLPP3)基因与鹅肥肝形成的关系,选取30只70日龄健康朗德鹅,随机分为填饲组与对照组,采用实时荧光定量PCR技术测定朗德鹅不同填饲阶段(填饲12和24 d)肝脏、胸肌和腹脂中PLPP3基因的表达水平;利用不同剂量的脂肪肝形成相关因子(胰岛素、葡萄糖、油酸、亚油酸和棕榈酸)分别处理朗德鹅原代肝细胞,检测这些因子对PLPP3基因表达的影响。结果表明:与对照组相比,填饲12 d时,肝脏、腹脂和胸肌中PLPP3基因的表达均显著或极显著下调;填饲24 d时,肝脏中表达差异不显著,腹脂中极显著下调,胸肌中显著上调;在鹅原代肝细胞中,与对照组相比,处理组葡萄糖、棕榈酸和油酸能显著抑制PLPP3基因的表达,而胰岛素和亚油酸处理则无显著效应。这一研究提示PLPP3基因的表达变化与鹅肥肝形成密切相关,且受脂肪形成相关因子的调控,说明PLPP3可能在鹅肥肝形成中有重要作用。
This study was to explore the relationship between PLPP3 gene and the formation of goose fatty liver, and to lay a foundation for its functional research. Thirty 70-day-old healthy Landaise geese were randomly divided into the overfeeding group and the control group(feeding ad libitum). The expression levels of PLPP3 gene in liver, pectoral muscle and abdominal fat of Landaise geese at different feeding stages(12 and 24 days) were determined by real-time fluorescence quantitative PCR. The primary hepatocytes of Landaise geese were treated with different doses of fatty liver-related factors(insulin, glucose, oleic acid, linoleic acid and palmitic acid) to detect the effects of these factors on the expression of PLPP3 gene. The results showed that compared with the control group, the expression of PLPP3 gene in the liver, abdominal fat and pectoral muscle decreased significantly(P<0.05) or very significantly(P<0.01) at 12 d of overfeeding. However, at 24 d of overfeeding, the expression of PLPP3 gene in the liver was similar to that in the control group, while the expression of the gene increased very significantly in pectoral muscle(P<0.01) and decreased significantly in abdominal fat(P<0.05). Glucose, palmitic acid and oleic acid could significantly inhibit the expression of PLPP3 gene in primary goose hepatocytes compared with control group(P< 0.05), while insulin and linoleic acid had no significant effect. The results showed that the expression of PLPP3 gene was closely related to the formation of goose fatty liver, and regulated by fatty liver-related factors, suggesting that PLPP3 may play an important role in the formation of goose fatty liver.
This study was to explore the relationship between PLPP3 gene and the formation of goose fatty liver, and to lay a foundation for its functional research. Thirty 70-day-old healthy Landaise geese were randomly divided into the overfeeding group and the control group(feeding ad libitum). The expression levels of PLPP3 gene in liver, pectoral muscle and abdominal fat of Landaise geese at different feeding stages(12 and 24 days) were determined by real-time fluorescence quantitative PCR. The primary hepatocytes of Landaise geese were treated with different doses of fatty liver-related factors(insulin, glucose, oleic acid, linoleic acid and palmitic acid) to detect the effects of these factors on the expression of PLPP3 gene. The results showed that compared with the control group, the expression of PLPP3 gene in the liver, abdominal fat and pectoral muscle decreased significantly(P<0.05) or very significantly(P<0.01) at 12 d of overfeeding. However, at 24 d of overfeeding, the expression of PLPP3 gene in the liver was similar to that in the control group, while the expression of the gene increased very significantly in pectoral muscle(P<0.01) and decreased significantly in abdominal fat(P<0.05). Glucose, palmitic acid and oleic acid could significantly inhibit the expression of PLPP3 gene in primary goose hepatocytes compared with control group(P< 0.05), while insulin and linoleic acid had no significant effect. The results showed that the expression of PLPP3 gene was closely related to the formation of goose fatty liver, and regulated by fatty liver-related factors, suggesting that PLPP3 may play an important role in the formation of goose fatty liver.
引文
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[16] 柳序,郭松长,刘耀文,等.鹅肝脏脂肪变性和鹅肥肝形成的分子保护机制研究进展 [J].动物营养学报,2018,30(7):2453-2458.
[17] BAE Y J,KIM S E,HONG S Y,et al.Time-course microarray analysis for identifying candidate genes involved in obesity-associated pathological changes in the mouse colon [J].Genes & Nutrition,2016,11(1):30-41.
[18] 李辉.Lipid Phosphate Phosphatase 3 (PLPP3)基因多态性与环境因素相互作用在脑血管狭窄中的作用 [D].青岛:青岛大学,2018.
[19] 王佳茜.神经酰胺在非酒精性脂肪性肝病中的应用研究进展 [J].现代中西医结合杂志,2016,25(33):3760-3762.
[20] ROHRBACH T,MACEYKA M,SPIEGEL S.Sphingosine kinase and sphingosine-1-phosphate in liver pathobiology [J].Critical Reviews in Biochemistry and Molecular Biology,2017,32:1-11.
[21] SAHINI N,BORLAK J.Genomics of human fatty liver disease reveal mechanistically linked lipid droplet-associated gene regulations in blunt steatosis and nonalcoholic steatohepatitis [J].Translational Research,2016,177:41-69.
[2] 薛启平.浅议鹅肥肝生产技术 [J].山东畜牧兽医,2017,38(3):69-70.
[3] 彭祥伟,王小军,布丽君,等.人工填饲朗德鹅鹅肥肝营养成分分析 [J].农产品加工,2016(19):33-35.
[4] 杨彪.脂联素受体对鹅肥肝的保护作用及其表达调控研究 [D].扬州:扬州大学,2016.
[5] 刘龙,王倩,许程,等.鹅肥肝形成中补体受体1基因的表达和调控研究 [J].中国家禽,2016,38(24):5-10.
[6] 房志家,陈婷,赵敏,等.磷脂酸磷酸酶与中性脂肪代谢调控 [J].生命科学,2013,25(7):669-675.
[7] 李一路,张晴晴,胡卫芹,等.磷脂酸磷酸酶在脂质代谢和信号转导中的作用及其调控 [J].植物生理学报,2017,53(6):897-904.
[8] 房志家.PAP为核心的三酰甘油合成及相关调控机制的研究 [D].上海:东华大学,2013.
[9] TOUATHAMICI Z,WEIDMANN H,BLUM Y,et al.Role of lipid phosphate phosphatase 3 in human aortic endothelial cell function [J].Cardiovascular Research,2017,112(3):702.
[10] BUSNELLI M,MANZINI S,PAROLINI C,et al.Lipid phosphate phosphatase 3 in vascular pathophysiology [J].Atherosclerosis,2018,271:156-165.
[11] WU C,HUANG R T,KUO C H,et al.Mechano-sensitive PLPP3 regulates endothelial responses to athero-relevant hemodynamic forces [J].Circulation Research,2015,117(4):446-469.
[12] RESCHEN M E,GAULTON K J,LIN D,et al.Lipid-induced epigenomic changes in human macrophages identify a coronary artery disease-associated variant that regulates PLPP3 expression through altered C/EBP-beta binding [J].Plos Genetics,2015,11(4):e1005061.
[13] WESTCOTT J M,PRECHTL A M,MAINE E A,et al.An epigenetically distinct breast cancer cell subpopulation promotes collective invasion [J].Journal of Clinical Investigation,2015,125(5):1927-1943.
[14] 洪胜辉,张军,邵丹,等.鹅肝细胞的高纯分离及培养 [J].中国兽医学报,2012,32(9):1378-1380.
[15] 柳序,刘耀文,匡佑华,等.鹅肥肝的形成及主要影响因素的研究进展 [J].经济动物学报,2019,23(1):1-5.
[16] 柳序,郭松长,刘耀文,等.鹅肝脏脂肪变性和鹅肥肝形成的分子保护机制研究进展 [J].动物营养学报,2018,30(7):2453-2458.
[17] BAE Y J,KIM S E,HONG S Y,et al.Time-course microarray analysis for identifying candidate genes involved in obesity-associated pathological changes in the mouse colon [J].Genes & Nutrition,2016,11(1):30-41.
[18] 李辉.Lipid Phosphate Phosphatase 3 (PLPP3)基因多态性与环境因素相互作用在脑血管狭窄中的作用 [D].青岛:青岛大学,2018.
[19] 王佳茜.神经酰胺在非酒精性脂肪性肝病中的应用研究进展 [J].现代中西医结合杂志,2016,25(33):3760-3762.
[20] ROHRBACH T,MACEYKA M,SPIEGEL S.Sphingosine kinase and sphingosine-1-phosphate in liver pathobiology [J].Critical Reviews in Biochemistry and Molecular Biology,2017,32:1-11.
[21] SAHINI N,BORLAK J.Genomics of human fatty liver disease reveal mechanistically linked lipid droplet-associated gene regulations in blunt steatosis and nonalcoholic steatohepatitis [J].Translational Research,2016,177:41-69.