2022, 42(5): 110-115. doi: 10.3969/j.issn.1673-9159.2022.05.014
关键词： 盐度驯化 , 再生鳞 , 虹鳟 , MMPs , 基因表达
Keywords： salinity acclimation , regenerated scales , Oncorhynchus mykiss , MMPs , gene expression
【目的】揭示MMP2 和MMP9 在虹鳟（Oncorhynchus mykiss）幼鱼鳞片再生过程中的作用及盐度驯化对鳞片再生的影响。【方法】基于虹鳟转录组数据提取MMP2 和MMP9 基因序列信息，基于二基因氨基酸序列进行MMPs基因的系统进化分析；设置盐度驯化组（盐度27）和对照组（盐度3），分别采集两组虹鳟幼鱼在鳞片再生过程中不同时间（1、3、7、14、21 d）的正常鳞和再生鳞样品，利用实时荧光定量PCR 技术检测MMP2 和MMP9 基因的表达水平。【结果】系统发育树分析表明，虹鳟MMP2 和MMP9 均与大西洋鲑（Salmo salar）相应MMPs 先聚为一支，再与其他鱼类MMPs聚为硬骨鱼类分支；对照组再生鳞MMP2和MMP9表达水平均总体呈先升后降趋势，在7 d时表达量最高；盐度驯化组再生鳞MMP2 基因表达水平变化趋势为先升后降，在3 d 时表达量达到最大值，而MMP9基因的表达水平呈降低趋势。【结论】MMP2 和MMP9 在鳞片再生过程中基因表达水平的变化可作为成骨细胞和破骨细胞增殖分化的特征之一；盐度驯化抑制了虹鳟鳞片再生过程中MMP2 和MMP9 表达水平，表明盐度变化对骨再生过程有抑制作用。
【目的】揭示MMP2 和MMP9 在虹鳟（Oncorhynchus mykiss）幼鱼鳞片再生过程中的作用及盐度驯化对鳞片再生的影响。【方法】基于虹鳟转录组数据提取MMP2 和MMP9 基因序列信息，基于二基因氨基酸序列进行MMPs基因的系统进化分析；设置盐度驯化组（盐度27）和对照组（盐度3），分别采集两组虹鳟幼鱼在鳞片再生过程中不同时间（1、3、7、14、21 d）的正常鳞和再生鳞样品，利用实时荧光定量PCR 技术检测MMP2 和MMP9 基因的表达水平。【结果】系统发育树分析表明，虹鳟MMP2 和MMP9 均与大西洋鲑（Salmo salar）相应MMPs 先聚为一支，再与其他鱼类MMPs聚为硬骨鱼类分支；对照组再生鳞MMP2和MMP9表达水平均总体呈先升后降趋势，在7 d时表达量最高；盐度驯化组再生鳞MMP2 基因表达水平变化趋势为先升后降，在3 d 时表达量达到最大值，而MMP9基因的表达水平呈降低趋势。【结论】MMP2 和MMP9 在鳞片再生过程中基因表达水平的变化可作为成骨细胞和破骨细胞增殖分化的特征之一；盐度驯化抑制了虹鳟鳞片再生过程中MMP2 和MMP9 表达水平，表明盐度变化对骨再生过程有抑制作用。
| [1] | BIRKEDAL-HANSEN H, MOORE W G I, BODDEN M K, et al.Matrix metalloproteinases:a review[J].Critical Reviews in Oral Biology&Medicine, 1993, 4(2):197-250. |
| [2] | FILANTI C, DICKSON G R, DI MARTINA D, et al.The Expression of metalloproteinase-2,-9, and-14 and of tissue inhibitors-1 and-2 is developmentally modulated during osteogenesis in vitro, the mature osteoblastic phenotype expressing metalloproteinase-14[J].Journal of Bone and Mineral Research, 2000, 15(11):2154-2168. |
| [3] | ENGSIG M T, CHEN Q J, VU T H, et al.Matrix metallo-proteinase 9 and vascular endothelial growth factor are essential for osteoclast recruitment into developing long bones[J].The Journal of Cell Biology, 2000, 151(4):879-890. |
| [4] | LV G, YANG S, CHEN, et al.MicroRNA-21 promotes bone mesenchymal stem cells migration in vitro by activating PI3K/Akt/MMPs pathway[J].Journal of Clinical Neuroscience, 2017, 46:156-162. |
| [5] | EVERTS V, DELAISSÉ J M, KORPER W, et al.Cysteine proteinases and matrix metalloproteinases play distinct roles in the subosteoclastic resorption zone[J].Journal of Bone and Mineral Research, 1998, 13(9):1420-1430. |
| [6] | MURAWALA H, PATEL S, RANADIVE I, et al.Variation in expression and activity pattern of mmp2 and mmp9 on different time scales in the regenerating caudal fin of Poecilia latipinna[J].Journal of Fish Biology, 2018, 92(5):1604-1619. |
| [7] | ANDO K, SHIBATA E, HANS S, et al.Osteoblast production by reserved progenitor cells in zebrafish bone regeneration and maintenance[J].Developmental Cell, 2017, 43(5):643-650. |
| [8] | PASQUALETTI S, CONGIU T, BANFI G, et al.Alendronate rescued osteoporotic phenotype in a model of glucocorticoidinduced osteoporosis in adult zebrafish scale[J].International Journal of Experimental Pathology, 2015, 96(1):11-20. |
| [9] | SHIMADA A, KAWANISHI T, KANEKO T, et al.Trunk exoskeleton in teleosts is mesodermal in origin[J].Nature Communications, 2013, 4(1):1-8. |
| [10] | BEREITER-HAHN J, ZYLBERBERG L.Regeneration of teleost fish scale[J].Comparative Biochemistry and Physiology Part A:Physiology, 1993, 105(4):625-641. |
| [11] | DE VRIEZE E, SHARIF F, METZ J R, et al.Matrix metalloproteinases in osteoclasts of ontogenetic and regenerating zebrafish scales[J].Bone, 2011, 48(4):704-712. |
| [12] | GUERREIRO P M, COSTA R, POWER D M.Dynamics of scale regeneration in seawater-and brackish water-acclimated sea bass, Dicentrarchus labrax[J].Fish Physiology and Biochemistry, 2013, 39(4):917-930. |
| [13] | ESMAEILI M, ABEDIAN KENARI A, ROMBENSO A N.Effects of fish meal replacement with meat and bone meal using garlic (Allium sativum) powder on growth, feeding, digestive enzymes and apparent digestibility of nutrients and fatty acids in juvenile rainbow trout (Oncorhynchus mykiss Walbaum, 1792)[J].Aquaculture Nutrition, 2017, 23(6):1225-1234. |
| [14] | 范兆廷,姜作发,韩英.冷水性鱼类养殖学[M]北京:中国农业出版社, 2008. |
| [15] | 刘文娟,董亢,刘骋跃,等.盐度驯化对虹鳟鳃,肌肉和肝脏非极性脂脂肪酸的影响[J].中国海洋大学学报(自然科学版), 2019, 49(3):72-78. |
| [16] | 付占斐,王鑫,王芳,等.不同盐度驯化方式对虹鳟和硬头鳟幼鱼存活和生长的影响[J].中国海洋大学学报(自然科学版), 2020, 50(7):37-43. |
| [17] | SHU B, ZHANG M, XIE R, et al.BMP2, but not BMP4, is crucial for chondrocyte proliferation and maturation during endochondral bone development[J].Journal of Cell Science, 2011, 124(20):3428-3440. |
| [18] | 曹晓颖,马晨夕,夏星,等.鳜颌骨重塑发育基因Bmp4的克隆与表达分析[J].基因组学与应用生物学, 2020, 39(11):5075-5083. |
| [19] | ISSA J P, GONZAGA M, KOTAKE B G, et al.Bone repair of critical size defects treated with autogenic, allogenic, or xenogenic bone grafts alone or in combination with rh BMP-2[J].Clinical Oral Implants Research, 2016, 27(5):558-566. |
| [20] | TEZUKA K, NEMOTO K, TEZUKA Y, et al.Identification of matrix metalloproteinase 9 in rabbit osteoclasts[J].Journal of Biological Chemistry, 1994, 269(21):15006-15009. |
| [21] | 贺静雅.斑马鱼尾鳍再生中相关基因的表达及血根碱对其再生过程的影响[D].新乡:河南师范大学, 2018. |
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