PROTEOMICS STUDY ON EFFECT OF BASIC FIBROBLAST GROWTH FACTOR LONG CIRCULATION LIPOSOME ON SPINAL CORD TRACTION INJURY IN RATS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

WANGWenyue ¹ , YANGTianfu ² ,  LIULei ² , PEIFuxing ²

1. Department of Orthopaedics, Guang'anmen Hospital, Chinese Academy of Medicine Science, Beijing, 100053, P. R. China;
2. Department of Orthopaedics, West China Hospital, Sichuan University;

Corresponding author：

LIULei, Email: liuinsistence@163.com

Keywords：

Spinal cord traction injury; Proteomics; Basic fibroblast growth factor; Long circulation liposome; Nano ultra-high performance liquid chromatography-electrospray tandem mass spectrometry; Rat

DOI：

10.7507/1002-1892.20140303

Video：

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Abstract Full text Figures/Tables Video References Cited by

Objective To explore the possible active mechanism of the basic fibroblast growth factor (bFGF) long circulation l iposome (LCL) (bFGF+LCL) on spinal cord traction injury in rats at the level of proteomics. Methods Twenty Sprague Dawly rats were randomly divided into groups A and B, 10 rats in each group. The models of spinal cord traction injury was established at T12-L3 spines. The rats were not treated in group A, and the rats were treated with bFGF+LCL (20μg/ kg) in group B. At 3 weeks after operation, the rats were sacrificed for harvesting T13-L2 spinal tissue specimens. The protein was extracted and quantified in the spinal tissue firstly. The proteins from spinal tissue were separated by two-dimensional gel electrophoresis and identified by mass spectrometry. The different expression profiling was established in each group, and the differentially expressed protein was determined by comparing the level of each spot with gel imaging software and manually. The proteins were identified by nano ultra-high performance liquid chromatography-electrospray tandem mass spectrometry (NanoUPLC-ESI-MS/MS), and the proteins were classified. Results The differentially expressed protein spots were found in 2 groups. Compared with group A, 4 spots were up-regulated and 6 were down-regulated in group B. NanoUPLC-ESI-MS/MS results showed that 18 significant proteins were identified in 26 differentially expressed proteins, including 4 apoptosis-related proteins, 3 nerve signal transduction related proteins, 7 proteins involved in metabolism, 1 unknown function protein, and 3 unnamed proteins. Conclusion The differentially expressed proteins are found in spinal cord traction injury of rats treated with bFGF+LCL. bFGF+LCL can affect the proteins expression in rats with spinal cord traction injury. The possible active mechanism is that it has protective and repair effects on injured spinal cord by nerve signal transduction, and regulation of nerve cells apoptosis and metabolism.

Citation： WANGWenyue, YANGTianfu, LIULei, PEIFuxing. PROTEOMICS STUDY ON EFFECT OF BASIC FIBROBLAST GROWTH FACTOR LONG CIRCULATION LIPOSOME ON SPINAL CORD TRACTION INJURY IN RATS. Chinese Journal of Reparative and Reconstructive Surgery, 2014, 28(11): 1397-1405. doi: 10.7507/1002-1892.20140303 Copy

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2.	王鸿丽.急性脊髓损伤及电针治疗大鼠的比较蛋白质组研究.北京:中国人民解放军军事医学科学院, 2009.
3.	黄亚东, 项琪, 张宏波, 等.碱性成纤维细胞生长因子脂质体的制备和评价.中国药学杂志, 2008, 43(13):992-997.
4.	Dos Santos N, Allen C, Doppen AM, et al. Influence of poly(ethylene glycol)grafting density and polymer length on liposomes:relating plasma circulation lifetimes to protein binding. Biochim Biophys Acta, 2007, 1768(6):1367-1377.
5.	王文岳, 杨天府, 刘雷, 等.一种新型脊柱撑开器的研制及脊髓牵张性损伤动物模型的建立.中国修复重建外科杂志, 2011, 25(6):705-710.
6.	Yaksh TL, Stevens CW. Simple catheter preparation for permitting bolus intrathecal administration during chronic intrathecal infusion. Pharmacol Biochem Behav, 1986, 25(2):483-485.
7.	Fisher M, Meadows ME, Tuyen D, et al. Delayed treatment with intravenous basic fibroblast growth factor reduces infarct size following permanent focal cerebral ischemia in rats. J Cereb FlowMetab, 1995, 15(6):953-959.
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9.	Crommelin DJA, Daemen T, Scherphof GL, et al. Liposomes:vehicles for the targeted and controlled del ivery of peptides and proteins. J Control Release, 1997, 46(122):165-175.
10.	Pan W, Kastin AJ. Penetration of neurotrophins and cytoltines across the blood-brain/blood-spinal cord barrier. Adv Drug Deliv Rev, 1999, 36(2-3):291-298.
11.	王文岳, 杨天府, 刘雷, 等.大鼠急性脊髓牵张损伤相关蛋白质的筛选和鉴定.中华外科杂志, 2012, 50(8):737-743.
12.	De Nicola AF, Gonzalez SL, Labombarda F, et al. Progesterone treatment of spinal cord injury. J Mol Neurosci, 2006, 28(1):3-15.
13.	沈正祥, 吕红斌, 李小明, 等.川芎嗪对大鼠急性脊髓损伤模型caspase-3和NF表达的影响.中南大学学报:医学版, 2008, 33(8):693-699.
14.	谢玲林, 胡火珍, 杨明理, 等.成年大鼠急性脊髓全横断损伤后差异表达蛋白的蛋白组学研究.四川大学学报:自然科学版, 2008, 45(2): 419-425.
15.	Dimlich RV, Showers MJ, Shi pley MT. Densitometric analysis of cytochrome oxidase in ischemic rat brain. Brain Res, 1990, 516(2):181-191.
16.	谭小玲, 柳君泽, 曹利飞, 等.长时间缺氧对大鼠脑皮质线粒体细胞色素氧化酶活性及亚基Ⅰ、Ⅳ蛋白表达的影响.中华航空航天医学杂志, 2002, 13(4):243-246.
17.	Nakatsuka H, Ohta S, Tanaka J, et al. Histochemical cytochrome coxidase activity and caspase-3 in gerbil hippocampal CA1 neurons after transient forebrain ischemia. Neurosci Lett, 2000, 285(2):127-130.
18.	Soane L, Kahraman S, Kristian T, et al. Mechanisms of impaired mitochondrial energy metabolism in acute and chronic neurodegenerative disorders. J Neurosci Res, 2007, 85(15):3407-3415.
19.	Ball RL, Albrecht T, Thompson WC, et al. Thrombin, EGF and phorbol myristate acetate stimulate tubul in polymerization in quiescent cells:A potential link to mitogenesis. Cell Motil Cytoskeleton, 1992, 23(4):265-278.
20.	Johnsen JI, Aurelio ON, Kwaja Z, et al. p53-mediated negative regulation of stathmin/Op18 expression is associated with G(2)/M cell-cycle arrest. Int J Cancer, 2000, 88(5):685-691.
21.	Manceau V, Gavet O, Curmi P, et al. Stathmin interaction with HSC70 family proteins. Electrophoresis, 1999, 20(2):409-417.
22.	Poulsen K, Bahl JM, Simonsen AH. Distinct transthyretin oxidation isoform profile in spinal fluid from patients with Alzheimer's disease and mild cognitive impairment. Clin Proteomics, 2014, 11(1):12.
23.	任惠民, 周志刚, 陈向军, 等.重症肌无力患者骨骼肌肉蛋白成分分析.中华神经科杂志, 2000, 33(4):227-230.
24.	Cabiscol E, Levine RL. The phosphatase activity of carbonic anhydraseⅢis reversibly regulated by glutathiolation. Proc Natl Acad Sic U S A, 1996, 93(9):4170-4174.
25.	Liu FT, Rabinovich GA. Galectins:regulators of acute and chronic inflammation. Ann N Y Acad Sci, 2010, 1183:158-182.
26.	Mendez-Huergo SP, Maller SM, Farez MF, et al. Integration of lectinglycan recognition systems and immune cell networks in CNS inflammation. Cytokine Growth Factor Rev, 2014, 25(3):247-255.
27.	Dianzani I, Howells DW, Ponzone A, et al. Two new mutations in the dihydropteridine reductase gene in patients with tetrahydrobiopterin deficiency. J Med Genet, 1993, 30(6):465-469.
28.	Gu Y, Gong Y, Zhang H, et al. Regulation of transforming growth factor beta 1 gene expression by dihydropteridine reductase in kidney 293T cells. Biochem Cell Biol, 2013, 91(3):187-193.
29.	Wood ZA, Schröder E, Robin Harris J, et al. Structure, mechanism and regulation of peroxiredoxins. Trends Biochem Sci, 2003, 28(1):32-40.
30.	Kayashima Y, Yamakawa-Kobayashi K. Involvement of Prx3, a Drosophila ortholog of the thiol-dependent peroxidase PRDX3, in agedependent oxidative stress resistance. Biomed Res, 2012, 33(5):319-322.
31.	Pillay CS, Hofmeyr JH, Rohwer JM. The logic of kinetic regulation in the thioredoxin system. BMC Syst Biol, 2011, 5:15.
32.	Smith RW, Cash P, Ellefsen S, et al. Proteomic changes in the crucian carp brain during exposure to anoxia. Proteomics, 2009, 9(8):2217-2229.
33.	Ramana KV, Chandra D, Srivastava S, et al. Aldose reductase mediates mitogenic signaling in vascular smooth muscle cells. J Biol Chem, 2002, 277(35):32063-32070.
34.	Keith RJ, Haberzettl P, Vladykovskaya E, et al. Aldose reductase decreases endop lasmic reticulum stress in ischemic hearts. Chem Biol Interact, 2009, 178(1-3):242-249.
35.	Tammali R, Ramana KV, Singhal SS, et al. Aldose reductase regulates growth factor-induced cyclooxygenase-2 expression and prostaglandin E2 production in human colon cancer cells. Cancer Res, 2006, 66(19): 9705-9713.
36.	Pandey S, Srivastava SK, Ramana KV. A potential therapeutic role for aldose reductase inhibitors in the treatment of endotoxin-related inflammatory diseases. Expert Opin Investig Drugs, 2012, 21(3):329-339.
37.	陈强, 陆江阳, 陆兵勋, 等.环氧合酶-2/CNPase对海洛因海绵状白质脑病中少突胶质细胞凋亡的作用.中华医学杂志, 2008, 88(25):857-858.
38.	Gravel M, Robert F, Kottis V, et al. 2', 3'-cycl ic nucleotide 3'-phosphodiesterase: a novel RNA-binding protein that inhibits protein synthesis. J Neurosci Res, 2009, 87(5):1069-1079.

1. Mugabe C, Azghani AO, Omri A. Li posome-mediated gentamicin delivery:development and activity against resistant strains of Pseudomonas aeruginosa isolated from cystic fibrosis patients. J Antimicrob-Chemother, 2005, 55(2):269-271.
2. 王鸿丽.急性脊髓损伤及电针治疗大鼠的比较蛋白质组研究.北京:中国人民解放军军事医学科学院, 2009.
3. 黄亚东, 项琪, 张宏波, 等.碱性成纤维细胞生长因子脂质体的制备和评价.中国药学杂志, 2008, 43(13):992-997.
4. Dos Santos N, Allen C, Doppen AM, et al. Influence of poly(ethylene glycol)grafting density and polymer length on liposomes:relating plasma circulation lifetimes to protein binding. Biochim Biophys Acta, 2007, 1768(6):1367-1377.
5. 王文岳, 杨天府, 刘雷, 等.一种新型脊柱撑开器的研制及脊髓牵张性损伤动物模型的建立.中国修复重建外科杂志, 2011, 25(6):705-710.
6. Yaksh TL, Stevens CW. Simple catheter preparation for permitting bolus intrathecal administration during chronic intrathecal infusion. Pharmacol Biochem Behav, 1986, 25(2):483-485.
7. Fisher M, Meadows ME, Tuyen D, et al. Delayed treatment with intravenous basic fibroblast growth factor reduces infarct size following permanent focal cerebral ischemia in rats. J Cereb FlowMetab, 1995, 15(6):953-959.
8. Ramagli LS. Quantifying protein in 2-D PAGE solubil ization buffers. Methods Mol Biol, 1999, 112:99-103.
9. Crommelin DJA, Daemen T, Scherphof GL, et al. Liposomes:vehicles for the targeted and controlled del ivery of peptides and proteins. J Control Release, 1997, 46(122):165-175.
10. Pan W, Kastin AJ. Penetration of neurotrophins and cytoltines across the blood-brain/blood-spinal cord barrier. Adv Drug Deliv Rev, 1999, 36(2-3):291-298.
11. 王文岳, 杨天府, 刘雷, 等.大鼠急性脊髓牵张损伤相关蛋白质的筛选和鉴定.中华外科杂志, 2012, 50(8):737-743.
12. De Nicola AF, Gonzalez SL, Labombarda F, et al. Progesterone treatment of spinal cord injury. J Mol Neurosci, 2006, 28(1):3-15.
13. 沈正祥, 吕红斌, 李小明, 等.川芎嗪对大鼠急性脊髓损伤模型caspase-3和NF表达的影响.中南大学学报:医学版, 2008, 33(8):693-699.
14. 谢玲林, 胡火珍, 杨明理, 等.成年大鼠急性脊髓全横断损伤后差异表达蛋白的蛋白组学研究.四川大学学报:自然科学版, 2008, 45(2): 419-425.
15. Dimlich RV, Showers MJ, Shi pley MT. Densitometric analysis of cytochrome oxidase in ischemic rat brain. Brain Res, 1990, 516(2):181-191.
16. 谭小玲, 柳君泽, 曹利飞, 等.长时间缺氧对大鼠脑皮质线粒体细胞色素氧化酶活性及亚基Ⅰ、Ⅳ蛋白表达的影响.中华航空航天医学杂志, 2002, 13(4):243-246.
17. Nakatsuka H, Ohta S, Tanaka J, et al. Histochemical cytochrome coxidase activity and caspase-3 in gerbil hippocampal CA1 neurons after transient forebrain ischemia. Neurosci Lett, 2000, 285(2):127-130.
18. Soane L, Kahraman S, Kristian T, et al. Mechanisms of impaired mitochondrial energy metabolism in acute and chronic neurodegenerative disorders. J Neurosci Res, 2007, 85(15):3407-3415.
19. Ball RL, Albrecht T, Thompson WC, et al. Thrombin, EGF and phorbol myristate acetate stimulate tubul in polymerization in quiescent cells:A potential link to mitogenesis. Cell Motil Cytoskeleton, 1992, 23(4):265-278.
20. Johnsen JI, Aurelio ON, Kwaja Z, et al. p53-mediated negative regulation of stathmin/Op18 expression is associated with G(2)/M cell-cycle arrest. Int J Cancer, 2000, 88(5):685-691.
21. Manceau V, Gavet O, Curmi P, et al. Stathmin interaction with HSC70 family proteins. Electrophoresis, 1999, 20(2):409-417.
22. Poulsen K, Bahl JM, Simonsen AH. Distinct transthyretin oxidation isoform profile in spinal fluid from patients with Alzheimer's disease and mild cognitive impairment. Clin Proteomics, 2014, 11(1):12.
23. 任惠民, 周志刚, 陈向军, 等.重症肌无力患者骨骼肌肉蛋白成分分析.中华神经科杂志, 2000, 33(4):227-230.
24. Cabiscol E, Levine RL. The phosphatase activity of carbonic anhydraseⅢis reversibly regulated by glutathiolation. Proc Natl Acad Sic U S A, 1996, 93(9):4170-4174.
25. Liu FT, Rabinovich GA. Galectins:regulators of acute and chronic inflammation. Ann N Y Acad Sci, 2010, 1183:158-182.
26. Mendez-Huergo SP, Maller SM, Farez MF, et al. Integration of lectinglycan recognition systems and immune cell networks in CNS inflammation. Cytokine Growth Factor Rev, 2014, 25(3):247-255.
27. Dianzani I, Howells DW, Ponzone A, et al. Two new mutations in the dihydropteridine reductase gene in patients with tetrahydrobiopterin deficiency. J Med Genet, 1993, 30(6):465-469.
28. Gu Y, Gong Y, Zhang H, et al. Regulation of transforming growth factor beta 1 gene expression by dihydropteridine reductase in kidney 293T cells. Biochem Cell Biol, 2013, 91(3):187-193.
29. Wood ZA, Schröder E, Robin Harris J, et al. Structure, mechanism and regulation of peroxiredoxins. Trends Biochem Sci, 2003, 28(1):32-40.
30. Kayashima Y, Yamakawa-Kobayashi K. Involvement of Prx3, a Drosophila ortholog of the thiol-dependent peroxidase PRDX3, in agedependent oxidative stress resistance. Biomed Res, 2012, 33(5):319-322.
31. Pillay CS, Hofmeyr JH, Rohwer JM. The logic of kinetic regulation in the thioredoxin system. BMC Syst Biol, 2011, 5:15.
32. Smith RW, Cash P, Ellefsen S, et al. Proteomic changes in the crucian carp brain during exposure to anoxia. Proteomics, 2009, 9(8):2217-2229.
33. Ramana KV, Chandra D, Srivastava S, et al. Aldose reductase mediates mitogenic signaling in vascular smooth muscle cells. J Biol Chem, 2002, 277(35):32063-32070.
34. Keith RJ, Haberzettl P, Vladykovskaya E, et al. Aldose reductase decreases endop lasmic reticulum stress in ischemic hearts. Chem Biol Interact, 2009, 178(1-3):242-249.
35. Tammali R, Ramana KV, Singhal SS, et al. Aldose reductase regulates growth factor-induced cyclooxygenase-2 expression and prostaglandin E2 production in human colon cancer cells. Cancer Res, 2006, 66(19): 9705-9713.
36. Pandey S, Srivastava SK, Ramana KV. A potential therapeutic role for aldose reductase inhibitors in the treatment of endotoxin-related inflammatory diseases. Expert Opin Investig Drugs, 2012, 21(3):329-339.
37. 陈强, 陆江阳, 陆兵勋, 等.环氧合酶-2/CNPase对海洛因海绵状白质脑病中少突胶质细胞凋亡的作用.中华医学杂志, 2008, 88(25):857-858.
38. Gravel M, Robert F, Kottis V, et al. 2', 3'-cycl ic nucleotide 3'-phosphodiesterase: a novel RNA-binding protein that inhibits protein synthesis. J Neurosci Res, 2009, 87(5):1069-1079.

Chinese Journal of Reparative and Reconstructive Surgery

PROTEOMICS STUDY ON EFFECT OF BASIC FIBROBLAST GROWTH FACTOR LONG CIRCULATION LIPOSOME ON SPINAL CORD TRACTION INJURY IN RATS

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