SUMO1和SENP1在新生大鼠肺发育过程中的表达
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摘要
目的探讨大鼠肺发育过程中SUMO1、SENP1表达变化及其与肺发育的关系。方法新生大鼠随机分为4组(n=8),分别于生后1、4、7和14 d处死取肺组织,用RT-qPCR检测肺组织SUMO1、SENP1 mRNA表达,Western blot检测肺组织SUMO1、SENP1蛋白表达。结果生后14 d内SUMO1 mRNA稳定表达; 14 d较4 d游离SUMO1表达减少,SUMO1结合蛋白表达增加(P<0. 05);生后4 d游离SUMO1蛋白表达较生后1及7 d均明显升高(P<0. 05),于生后7和14 d表达趋于稳定; SENP1 mRNA及蛋白表达变化与游离SUMO1变化趋势一致。结论 SENP1通过介导蛋白去SUMO1修饰,维持蛋白SUMO化动态平衡,这一平衡调节可能在肺发育过程中起重要作用。
Objective To investigate the expression of SUMO1 and SENP1 during lung development in rats.Methods Neonatal rats were randomly divided into 4 groups with 8 rats in each. Pulmonary tissue was collected for further study at 1,4,7,14 d after birth. The mRNA of SUMO1 and SENP1 was detected by RT-qPCR. Western blot was used to analyze the protein expression of SUMO1 and SENP1. Results The expression of SUMO1 mRNA had no significant changes at 1 to 14 d. Compared with 4 d,free SUMO1 decreased and conjugated SUMO1 increased at 14 d( P<0. 05). The protein expression of free SUMO1 at 4 d was significantly higher than 1 and 7 d( P<0. 05),then remained stable at 7 to 14 d. The expressions levels of SENP1 mRNA and protein showed the same tendency with free SUMO1. Conclusions SENP1 maintains the dynamic balance of SUMOylation by mediating deconjugation of SUMO1 protein,this process may play an important role in lung development.
Objective To investigate the expression of SUMO1 and SENP1 during lung development in rats.Methods Neonatal rats were randomly divided into 4 groups with 8 rats in each. Pulmonary tissue was collected for further study at 1,4,7,14 d after birth. The mRNA of SUMO1 and SENP1 was detected by RT-qPCR. Western blot was used to analyze the protein expression of SUMO1 and SENP1. Results The expression of SUMO1 mRNA had no significant changes at 1 to 14 d. Compared with 4 d,free SUMO1 decreased and conjugated SUMO1 increased at 14 d( P<0. 05). The protein expression of free SUMO1 at 4 d was significantly higher than 1 and 7 d( P<0. 05),then remained stable at 7 to 14 d. The expressions levels of SENP1 mRNA and protein showed the same tendency with free SUMO1. Conclusions SENP1 maintains the dynamic balance of SUMOylation by mediating deconjugation of SUMO1 protein,this process may play an important role in lung development.
引文
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[8]陈筱青,蒋炜,吴升华.晚期肺发育障碍与支气管肺发育不良研究进展[J].国际儿科学杂,2015,42:615-618.
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[11] Francisco JV,Noelia LP,Mario GD. The Sumo protease Senp7 is required for proper neuronal differentiation[J].Biochim Biophys Acta,2016,1863:1490-1498.
[12] Tan F,Dong W,Lei X,et al. Attenuated SUMOylation of sirtuin 1 in premature neonates with bronchopulmonary dysplasia[J]. Mol Med Rep,2018,17:1283-1288.
[13] Yuasa E,Saitoh H. In situ SUMOylation and De SUMOylation assays:fluorescent methods to visualize SUMOylation and De SUMOylation in permeabilized cells[J]. Methods Mol Biol,2016,1475:151-159.
[2] Nuro-Gyina PK,Parvin JD. Roles for SUMO in premRNA processing[J]. Wiley Interdiscip Rev RNA,2015,7:105-112.
[3] Chen YD,Liu JY,Lu YM,et al. Functional roles of C/EBPαand SUMO-modification in lung development[J].Int J Mol Med,2017,40:1037-1046.
[4] Saho E,Takuya A,Hiroshi A,et al. The SUMO protease SENP1 is required for cohesion maintenance and mitotic arrest following spindle poison treatment[J]. Biochem Biophys Res Commun,2012,426:310-316.
[5] Chen CH,Chang CC,Lee TH,et al. SENP1 de SUMOylates and regulates Pin1 protein activity and cellular function[J]. Cancer Res,2013,73:3951-3962.
[6]田华,戴爱国,符代炎,等. SENP1在大鼠低氧性肺动脉高压形成中肺血管壁中表达及可能作用[J].中国应用生理学杂志,2012,28:123-127.
[7]李珍,黄宇戈,肖芳芳.血管内皮生长因子和内皮抑素在大鼠肺的表达及其与肺发育的关系[J].中国新生儿科杂志,2015,30:138-142.
[8]陈筱青,蒋炜,吴升华.晚期肺发育障碍与支气管肺发育不良研究进展[J].国际儿科学杂,2015,42:615-618.
[9] LomelíH,Vzquez M. Emerging roles of the SUMO pathway in development[J]. Cell Mol Life Sci,2011,68:4045-4064.
[10] Sharma P,Yamada S,Lualdi M,et al. Senp1 is essential for desumoylating Sumo1-modified proteins but dispensable for Sumo2 and Sumo3 deconjugation in the mouse embryo[J]. Cell Rep,2013,3:1640-1650.
[11] Francisco JV,Noelia LP,Mario GD. The Sumo protease Senp7 is required for proper neuronal differentiation[J].Biochim Biophys Acta,2016,1863:1490-1498.
[12] Tan F,Dong W,Lei X,et al. Attenuated SUMOylation of sirtuin 1 in premature neonates with bronchopulmonary dysplasia[J]. Mol Med Rep,2018,17:1283-1288.
[13] Yuasa E,Saitoh H. In situ SUMOylation and De SUMOylation assays:fluorescent methods to visualize SUMOylation and De SUMOylation in permeabilized cells[J]. Methods Mol Biol,2016,1475:151-159.