电离辐照损伤对小鼠睾丸组织氧化磷酸化信号通路的影响
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摘要
目的比较电离辐照损伤小鼠与正常小鼠的睾丸组织差异表达蛋白质,分析所得差异蛋白涉及的信号通路,以期从蛋白质组学角度阐述电离辐照对睾丸组织损伤的作用机制。方法采用~(60)Coγ射线辐照建立小鼠电离辐照损伤模型。运用比较蛋白质组学方法、iTRAQ联合LC-MS/MS检测技术,提取出小鼠辐照组与正常组睾丸组织比较后的差异表达蛋白质,进一步利用David6.8、String10.5和Cytoscape3.6.1数据库,对差异蛋白进行KEGG富集分析和相互作用分析。结果 KEGG富集分析鉴定出21条生物信号通路(P<0.05),其中富集于氧化磷酸化信号通路的差异蛋白有13个,均为表达下调。该通路涉及ATP合成酶、细胞色素、NADH脱氢酶这3类蛋白的多个亚基,它们主要参与线粒体电子传递、线粒体呼吸链复合物I装配、ATP生物合成等过程。结论电离辐照引起小鼠睾丸组织氧化磷酸化信号通路所涉及的13个差异蛋白均出现低表达,导致细胞机体ATP的合成障碍是辐射损伤的重要机制。
Aim To compare the differential expression of proteins in testicular tissues of ionized irradiated mice and normal mice, in order to explain the mechanism of ionizing irradiation on testicular tissue damage from proteomics perspective. Methods The mouse ionizing radiation damage model was established by cobalt 60-ray irradiation. Comparative proteomics method, iTRAQ combined with LC-MS/MS detection technology were used to extract differentially expressed proteins of testis tissues from mouse irradiated group and normal group, then David6.8, String10.5 and Cytoscape 3.6.1 database were employed for KEGG enrichment analysis and interaction analysis on differential proteins. Results Twenty-one biological signaling pathways(P<0.05) were identified by KEGG enrichment analysis, with 13 differentially expressed proteins enriched in phosphoric oxide signaling pathways, and both were downgrades of expression. The pathway involved several subgroups of ATP synthase, cytochrome and NADH dehydrogenase, which were mainly involved in mitochondrial electron transfer, mitochondrial respiratory chain complex I assembly, ATP biosynthesis and so on. Conclusions Ionizing radiation is responsible for the expression of oxidative phosphorylation signaling pathway in mouse testis tissues. The low expression of 13 differential proteins leads to the synthesis of cell ATP. It is an important mechanism of radiation damage.
Aim To compare the differential expression of proteins in testicular tissues of ionized irradiated mice and normal mice, in order to explain the mechanism of ionizing irradiation on testicular tissue damage from proteomics perspective. Methods The mouse ionizing radiation damage model was established by cobalt 60-ray irradiation. Comparative proteomics method, iTRAQ combined with LC-MS/MS detection technology were used to extract differentially expressed proteins of testis tissues from mouse irradiated group and normal group, then David6.8, String10.5 and Cytoscape 3.6.1 database were employed for KEGG enrichment analysis and interaction analysis on differential proteins. Results Twenty-one biological signaling pathways(P<0.05) were identified by KEGG enrichment analysis, with 13 differentially expressed proteins enriched in phosphoric oxide signaling pathways, and both were downgrades of expression. The pathway involved several subgroups of ATP synthase, cytochrome and NADH dehydrogenase, which were mainly involved in mitochondrial electron transfer, mitochondrial respiratory chain complex I assembly, ATP biosynthesis and so on. Conclusions Ionizing radiation is responsible for the expression of oxidative phosphorylation signaling pathway in mouse testis tissues. The low expression of 13 differential proteins leads to the synthesis of cell ATP. It is an important mechanism of radiation damage.
引文
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[2] Kouvaris J R, Kouloulias V E, Vlahos L J. Amifostine: the fist selective–target and broad–spectrum radioprotector[J]. Oncologist , 2007, 12(6): 738-47.
[3] Kuntic V S, Stankovic M B, Vujic Z B, et al. Radioprotectors -the evergreen topic[J]. Chem Biodivers, 2013, 10(10): 1791–803.
[4] 刘方娟. 小鼠睾丸糖基化蛋白质组学研究[D]. 南京:南京医科大学,2016.[4] Liu F J. Mouse testis glycosylation proteomics study [D]. Nanjing: Nanjing Medical University, 2016.
[5] 仵春云,赵昌涛,张杜平,等.地黄多糖干预对低剂量X线照射小鼠生精细胞P53和Bcl-2蛋白表达的影响[J].中国男科学杂志, 2015,29(11):8-12.[5] Wu C Y,Zhao C T,Zhang D P, et al. Effects of Rehmannia polysaccharide intervention on expression of P53 and Bcl-2 protein in spermatogenic cells of low-dose X-ray irradiation mice[J]. Chin J Androl, 2015, 29(11): 8-12.
[6] Gao X F, Wang S M, Peng R Y.Status quo of the researches on the biological effect of electromagnetic radiation on the testis and epididymal sperm[J]. Natl J Androl, 2007, 13(9): 826-9.
[7] Fukunaga, Kaminaga K, Sato T, et al. Application of an ex vivo tissue model to investigate radiobiological effects on Spermatogenesis[J]. Radiat Res, 2018, 189(6): 661-7.
[8] Yamamori T, Yasui H, Yamazumi M, et al. Ionizing radiation induces mitochondrial reactive oxygen species production accompanied by upregulation of mitochondrial electron transport chain function and mitochondrial content under control of the cell cycle checkpoint[J]. Free Radic Biol Med, 2012, 53(2):260-70.
[9] Yamamori T, Ike S, Bo T, et al. Inhibition of the mitochondrial fission protein dynamin-related protein1 (Drp1) impairs mitochondrial fission and mitotic catastrophe after X-irradiation[J]. Mol Biol Cell, 2015, 26: 4607-17.
[10] Rajendran S, Harrison S H, Thomas R A, et al. The role of mitochondria in the radiation-induced bystander effect in human Iymphoblastoid cells[J]. Radiat Res, 2011, 175(2): 159-71.
[11] 李敏,赵领,刘真宏,等.川芎提取物对电离辐照损伤实验小鼠脾脏组织细胞凋亡相关蛋白表达的影响[J].中国药理学通报, 2018,34(11):1544-8.[11] Li M, Zhao L, Liu Z H, et al. Effects of Ligusticum chuanxiong Hort. on the expression of apoptosis-related proteins in spleen tissue of mice with ionizing radiation injury[J]. Chin Pharmacol Bull, 2018, 34(11): 1544-8.
[12] 朱文赫,沈楠,徐俊杰,等.微波辐射对大鼠能量代谢及心肌细胞凋亡的影响[J].中国病理生理杂志, 2015, 31(4): 647-51.[12] Zhu W H,Shen N,Xu J J ,et al.Effects of microwave radiation on energy metabolism and cardiomyocyte apoptosis in rats[J]. Chin J Pathophysiol, 2015, 31(4): 647-51.
[13] 王水明,王德文,彭瑞云,等.电磁脉冲辐射对小鼠生精细胞能量代谢的影响[J].中华物理医学与康复杂志, 2003, (9): 516-9.[13] Wang S M,Wang D W,Peng R Y,et al. Effect of electromagnetic pulse radiation on energy metabolism of spermatogenic cells in mice[J]. Chin J Physical Med Rehabilitation, 2003, (9): 516-9.
[14] 徐婷,李华,鲁姗姗,等. 线粒体电子传递呼吸链及其生物学意义的研究进展[J]. 复旦学报(医学版), 2015, 42(2): 250-5, 261.[14] Xu T, Li H, Lu S S, et al. Progress in mitochondrial electron transport respiratory chain and its biological significance[J]. J Fudan Univ (Med Sci), 2015, 42(2): 250-5, 261.
[15] 袁李佳龙, 秦旭平. ATP合酶β亚基功能与疾病[J]. 中国临床药理学与治疗学, 2018, 23(4): 464-70.[15] Yuan L J L,Qin X P. ATP synthase β subunit function and disease [J]. Chin J Clin Pharmacol Ther, 2018, 23(4): 464-70.