菲律宾蛤仔PPAR基因家族特征分析及干露胁迫下表达变化
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摘要
为研究干露胁迫下菲律宾蛤仔Ruditapes philippinarum脂质代谢相关基因PPAR所发挥的作用,对菲律宾蛤仔PPAR基因家族(PPARα、PPARβ、PPARγ)进行了鉴定及组织表达分析,并考察了低温(5℃)、常温(20℃)干露胁迫对菲律宾蛤仔PPAR基因表达变化的影响。结果表明:PPARα、PPARβ和PPARγ的开放阅读框长度分别为912 bp、1644 bp和2124 bp,分别编码了239、547、707个氨基酸;系统进化树显示,菲律宾蛤仔PPARα和PPARγ与虾夷扇贝Patinopecten yessoensis亲缘关系较近,PPARβ与福寿螺Pomacea canalicalata亲缘关系较近;荧光定量分析显示,菲律宾蛤仔脂质代谢相关基因PPARα/β/γ均可不同程度地响应干露胁迫;整体来看,干露胁迫会抑制PPARα及PPARγ表达,并且低温(5℃)干露会显著抑制二者表达(P<0.05),但会诱导PPARβ表达,并且20℃干露胁迫对菲律宾蛤仔机体内部脂质代谢平衡的影响显著高于5℃(P<0.05)。研究表明,干露胁迫下,菲律宾蛤仔PPAR基因家族参与调控脂肪酸氧化,以维持保蛤仔体内能量平衡,并且低温可减少干露胁迫下菲律宾蛤仔机体的能量消耗,有助于菲律宾蛤仔适应干露环境。
The identification, characterization and expression of PPAR gene family(PPARα, PPARβ, PPARγ) were investigated in tissues of Manila clam Ruditapes philippinarum with shell length of 2.5 cm-3.0 cm exposed to air at low temperature(5 ℃)and normal temperature(20 ℃)in order to evaluate the role of PPAR gene in Manila clam lipid metabolism under air exposure stress. It was found that the length of open reading frame was 912 bp, encoding 239 amino acids, in PPARα; 1644 bp, encoding 547 amino acids, in PPARβ and 2124 bp, encoding 707 amino acids, in PPARγ. The phylogenetic tree showed that PPARα and PPARγ had high homology in Manila clam with yesso scallop Patinopecten yessoensis; PPARβ with high homology to Pomacea canaliculata. Quantitative analysis revealed that PPARα/β/γ, a lipid metabolism-related gene, responded to air exposure stimulation to some extent, significant expression inhibition of PPARα and PPARγ by air exposure and low-temperature air exposure(P<0.05). However, the expression of PPARβ was found to be induced, and the effect of air exposure stress on lipid metabolism balance in the clam was significantly higher at 20 ℃ than that at 5 ℃(P<0.05). The findings indicate that the PPAR gene family of Manila clam is involved in regulation of fatty acid oxidation to maintain energy balance in the clam under air exposure, and that low temperature reduces energy consumption under air exposure, thus resulting in helpping the clam adapt to this environment.
The identification, characterization and expression of PPAR gene family(PPARα, PPARβ, PPARγ) were investigated in tissues of Manila clam Ruditapes philippinarum with shell length of 2.5 cm-3.0 cm exposed to air at low temperature(5 ℃)and normal temperature(20 ℃)in order to evaluate the role of PPAR gene in Manila clam lipid metabolism under air exposure stress. It was found that the length of open reading frame was 912 bp, encoding 239 amino acids, in PPARα; 1644 bp, encoding 547 amino acids, in PPARβ and 2124 bp, encoding 707 amino acids, in PPARγ. The phylogenetic tree showed that PPARα and PPARγ had high homology in Manila clam with yesso scallop Patinopecten yessoensis; PPARβ with high homology to Pomacea canaliculata. Quantitative analysis revealed that PPARα/β/γ, a lipid metabolism-related gene, responded to air exposure stimulation to some extent, significant expression inhibition of PPARα and PPARγ by air exposure and low-temperature air exposure(P<0.05). However, the expression of PPARβ was found to be induced, and the effect of air exposure stress on lipid metabolism balance in the clam was significantly higher at 20 ℃ than that at 5 ℃(P<0.05). The findings indicate that the PPAR gene family of Manila clam is involved in regulation of fatty acid oxidation to maintain energy balance in the clam under air exposure, and that low temperature reduces energy consumption under air exposure, thus resulting in helpping the clam adapt to this environment.
引文
[1] Tyagi S,Gupta P,Saini A S,et al.The peroxisome proliferator-activated receptor:a family of nuclear receptors role in various diseases[J].Journal of Advanced Pharmaceutical Technology & Research,2011,2(4):236-240.
[2] 张乾勇.PPAR的结构与功能及其生物学作用[J].国外医学卫生学分册,2000,27(5):284-288.
[3] 杨谷良,潘敏雄,向福,等.PPARγ调控脂肪细胞增殖和分化机理研究进展[J].食品科学,2017,38(3):254-260.
[4] 杨亚维,袁杰.PPARα抗氧化应激作用的研究进展[J].东南大学学报:医学版,2016,35(1):147-150.
[5] He Shan,Liang Xufang,Qu Chunmei,et al.Identification,organ expression and ligand-dependent expression levels of peroxisome proliferator activated receptors in grass carp(Ctenopharyngodon idella)[J].Comparative Biochemistry and Physiology Part C:Toxicology & Pharmacology,2012,155(2):381-388.
[6] 陈娜,吴英良.PPARδ的结构及其生物学功能与疾病[J].中国药理学通报,2006,22(9):1035-1038.
[7] 陈永熙,王伟铭,周同,等.PPAR-γ作用及其相关信号转导途径[J].细胞生物学杂志,2006,28(3):382-386.
[8] Yin Xuwang,Chen Peng,Chen Hai,et al.Physiological performance of the intertidal Manila clam (Ruditapes philippinarum) to long-term daily rhythms of air exposure[J].Scientific Reports,2017,DOI:https://doi.org/10.1038/srep41648.
[9] 闫喜武.菲律宾蛤仔养殖生物学、养殖技术与品种选育[D].青岛:中国科学院研究生院(海洋研究所),2005.
[10] 刘慧慧,周晏琳,张晴,等.菲律宾蛤仔捕后干露处置对其复水湿藏稳定性的影响[J].大连海洋大学学报,2018,33(2):244-250.
[11] 王文文.干露胁迫对菲律宾蛤仔免疫指标和基因表达的影响[D].大连:大连海洋大学,2016.
[12] 于瑞海,王昭萍,孔令锋,等.不同发育期的太平洋牡蛎在不同干露状态下的成活率研究[J].中国海洋大学学报,2006,36(4):617-620.
[13] 于瑞海,辛荣,赵强,等.海湾扇贝不同发育阶段耐干露的研究[J].海洋科学,2007,31(6):6-9.
[14] Guellich A,Damy T,Lecarpentier Y,et al.Role of oxidative stress in cardiac dysfunction of PPAR -/- mice[J].American Journal of Physiology-Heart and Circulatory Physiology,2007,293(1):H93-H102.
[15] Ali F,Nakamura K.Metabolic characteristics of the Japanese clam Ruditapes philippinarum (Adams & Reeve) during aerial exposure[J].Aquaculture Research,2000,31(2):157-165.
[16] Giordano Attianese G M P,Desvergne B.Integrative and systemic approaches for evaluating PPARβ/δ (PPARD) function[J].Nuclear Receptor Signaling,2015,13:1-32.
[17] Grimaldi P A.The roles of PPARs in adipocyte differentiation[J].Progress in Lipid Research,2001,40(4):269-281.
[18] Ibabe A,Bilbao E,Cajaraville M P.Expression of peroxisome proliferator-activated receptors in zebrafish (Danio rerio) depending on gender and developmental stage[J].Histochemistry and Cell Biology,2005,123(1):75-87.
[19] 贾成霞,张照斌,张清靖,等.虹鳟PPARα基因克隆、序列分析及其组织表达分布[J].中国水产科学,2012,19(4):707-714.
[20] You Cuihong,Jiang Danli,Zhang Qinghao,et al.Cloning and expression characterization of peroxisome proliferator-activated receptors (PPARs) with their agonists,dietary lipids,and ambient salinity in rabbitfish Siganus canaliculatus[J].Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology,2017,206:54-64.
[21] 林亚秋,吉红,郑玉才.草鱼PPARα和PPARγ基因的克隆与组织表达差异[J].水产科学,2011,30(2):94-97.
[22] Altschul S F,Gish W,Miller W,et al.Basic local alignment search tool[J].Journal of Molecular Biology,1990,215(3):403-410.
[23] 牟政强,闫路路,王化敏,等.菲律宾蛤仔不同发育时期及成体不同组织中内参基因筛选[J].水产学报,2018,42(5):663-672.
[24] 刘忠颖,刘卫东,李军,等.温度和规格对虾夷扇贝干露耐受性的影响[J].水产科学,2018,37(4):464-468.
[25] Gilde A J,van der Lee K A J M,Willemsen P H M,et al.Peroxisome proliferator-activated receptor (PPAR) α and PPARβ/δ,but not PPARγ,modulate the expression of genes involved in cardiac lipid metabolism[J].Circulation Research,2003,92(5):518-524.
[26] Luquet S,Lopez-Soriano J,Holst D,et al.Peroxisome proliferator-activated receptor δ controls muscle development and oxidative capability[J].FASEB Journal:Official Publication of the Federation of American Societies for Experimental Biology,2003,17(15):2299-2301.
[27] 林元烧,沈国英,张华.菲律宾蛤仔耗氧率的研究[J].厦门大学学报:自然科学版,1996,35(3):407-411.
[28] Marx N,Hombach V.Peroxisome proliferator-activated receptors (PPARs) in der Gef??w and-neue Regulatoren der Genexpression in vaskul?ren Zellen[J].Zeitschriftfü Kardiologie,2001,90(7):470-477.
[29] Barak Y,Nelson M C,Ong E S,et al.PPARγ is required for placental,cardiac,and adipose tissue development[J].Molecular Cell,1999,4(4):585-595.
[30] Momose H,Kaminuma T,Tanaka Y,et al.Analysis of gene regulation network by nuclear receptor PPAR[J].Genome Informatics,2003,14:364-365.
[31] 徐世文,樱桃,李术.PPAR-γ功能的研究进展[J].东北农业大学学报,2011,42(9):1-6.
[2] 张乾勇.PPAR的结构与功能及其生物学作用[J].国外医学卫生学分册,2000,27(5):284-288.
[3] 杨谷良,潘敏雄,向福,等.PPARγ调控脂肪细胞增殖和分化机理研究进展[J].食品科学,2017,38(3):254-260.
[4] 杨亚维,袁杰.PPARα抗氧化应激作用的研究进展[J].东南大学学报:医学版,2016,35(1):147-150.
[5] He Shan,Liang Xufang,Qu Chunmei,et al.Identification,organ expression and ligand-dependent expression levels of peroxisome proliferator activated receptors in grass carp(Ctenopharyngodon idella)[J].Comparative Biochemistry and Physiology Part C:Toxicology & Pharmacology,2012,155(2):381-388.
[6] 陈娜,吴英良.PPARδ的结构及其生物学功能与疾病[J].中国药理学通报,2006,22(9):1035-1038.
[7] 陈永熙,王伟铭,周同,等.PPAR-γ作用及其相关信号转导途径[J].细胞生物学杂志,2006,28(3):382-386.
[8] Yin Xuwang,Chen Peng,Chen Hai,et al.Physiological performance of the intertidal Manila clam (Ruditapes philippinarum) to long-term daily rhythms of air exposure[J].Scientific Reports,2017,DOI:https://doi.org/10.1038/srep41648.
[9] 闫喜武.菲律宾蛤仔养殖生物学、养殖技术与品种选育[D].青岛:中国科学院研究生院(海洋研究所),2005.
[10] 刘慧慧,周晏琳,张晴,等.菲律宾蛤仔捕后干露处置对其复水湿藏稳定性的影响[J].大连海洋大学学报,2018,33(2):244-250.
[11] 王文文.干露胁迫对菲律宾蛤仔免疫指标和基因表达的影响[D].大连:大连海洋大学,2016.
[12] 于瑞海,王昭萍,孔令锋,等.不同发育期的太平洋牡蛎在不同干露状态下的成活率研究[J].中国海洋大学学报,2006,36(4):617-620.
[13] 于瑞海,辛荣,赵强,等.海湾扇贝不同发育阶段耐干露的研究[J].海洋科学,2007,31(6):6-9.
[14] Guellich A,Damy T,Lecarpentier Y,et al.Role of oxidative stress in cardiac dysfunction of PPAR -/- mice[J].American Journal of Physiology-Heart and Circulatory Physiology,2007,293(1):H93-H102.
[15] Ali F,Nakamura K.Metabolic characteristics of the Japanese clam Ruditapes philippinarum (Adams & Reeve) during aerial exposure[J].Aquaculture Research,2000,31(2):157-165.
[16] Giordano Attianese G M P,Desvergne B.Integrative and systemic approaches for evaluating PPARβ/δ (PPARD) function[J].Nuclear Receptor Signaling,2015,13:1-32.
[17] Grimaldi P A.The roles of PPARs in adipocyte differentiation[J].Progress in Lipid Research,2001,40(4):269-281.
[18] Ibabe A,Bilbao E,Cajaraville M P.Expression of peroxisome proliferator-activated receptors in zebrafish (Danio rerio) depending on gender and developmental stage[J].Histochemistry and Cell Biology,2005,123(1):75-87.
[19] 贾成霞,张照斌,张清靖,等.虹鳟PPARα基因克隆、序列分析及其组织表达分布[J].中国水产科学,2012,19(4):707-714.
[20] You Cuihong,Jiang Danli,Zhang Qinghao,et al.Cloning and expression characterization of peroxisome proliferator-activated receptors (PPARs) with their agonists,dietary lipids,and ambient salinity in rabbitfish Siganus canaliculatus[J].Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology,2017,206:54-64.
[21] 林亚秋,吉红,郑玉才.草鱼PPARα和PPARγ基因的克隆与组织表达差异[J].水产科学,2011,30(2):94-97.
[22] Altschul S F,Gish W,Miller W,et al.Basic local alignment search tool[J].Journal of Molecular Biology,1990,215(3):403-410.
[23] 牟政强,闫路路,王化敏,等.菲律宾蛤仔不同发育时期及成体不同组织中内参基因筛选[J].水产学报,2018,42(5):663-672.
[24] 刘忠颖,刘卫东,李军,等.温度和规格对虾夷扇贝干露耐受性的影响[J].水产科学,2018,37(4):464-468.
[25] Gilde A J,van der Lee K A J M,Willemsen P H M,et al.Peroxisome proliferator-activated receptor (PPAR) α and PPARβ/δ,but not PPARγ,modulate the expression of genes involved in cardiac lipid metabolism[J].Circulation Research,2003,92(5):518-524.
[26] Luquet S,Lopez-Soriano J,Holst D,et al.Peroxisome proliferator-activated receptor δ controls muscle development and oxidative capability[J].FASEB Journal:Official Publication of the Federation of American Societies for Experimental Biology,2003,17(15):2299-2301.
[27] 林元烧,沈国英,张华.菲律宾蛤仔耗氧率的研究[J].厦门大学学报:自然科学版,1996,35(3):407-411.
[28] Marx N,Hombach V.Peroxisome proliferator-activated receptors (PPARs) in der Gef??w and-neue Regulatoren der Genexpression in vaskul?ren Zellen[J].Zeitschriftfü Kardiologie,2001,90(7):470-477.
[29] Barak Y,Nelson M C,Ong E S,et al.PPARγ is required for placental,cardiac,and adipose tissue development[J].Molecular Cell,1999,4(4):585-595.
[30] Momose H,Kaminuma T,Tanaka Y,et al.Analysis of gene regulation network by nuclear receptor PPAR[J].Genome Informatics,2003,14:364-365.
[31] 徐世文,樱桃,李术.PPAR-γ功能的研究进展[J].东北农业大学学报,2011,42(9):1-6.
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