退行性变腰椎间盘中基质细胞衍生因子1的表达及意义

doi:10.3969/j.issn.2095-4344.2013.24.017

摘要/Abstract

摘要：

背景：目前腰椎间盘退行性变确切的发病机制并不十分清楚。炎症参与腰椎间盘退行性变的发病机制，基质细胞衍生因子1属于趋化因子家族成员，与炎症有关。
目的：检测腰椎间盘退行性变患者椎间盘中基质细胞衍生因子1的表达水平，分析其与病情严重程度的关系。
方法：选取84例腰椎间盘退行性变患者和28例椎体爆裂性骨折患者，收集2组患者术后的椎间盘组织，用酶联免疫吸附的方法测定椎间盘中基质细胞衍生因子1的表达水平。根据Schneiderman标准进行分级，分析椎间盘中的基质细胞衍生因子1水平与疾病分级的关系。
结果与结论：与椎体爆裂性骨折患者相比，腰椎间盘退行性变患者的腰椎间盘组织中基质细胞衍生因子1水平明显升高，差异有显著性意义(P < 0.01)。Schneiderman 4级的患者椎间盘组织中基质细胞衍生因子1水平明显高于Schneiderman 2级和3级的患者，而Schneiderman 3级的患者椎间盘组织中基质细胞衍生因子1水平明显高于2级患者。另外，Spearman相关分析也显示，基质细胞衍生因子1的蛋白水平与Schneiderman分级呈正相关(r=0.412, P < 0.01)。提示腰椎间盘退行性变患者椎间盘中基质细胞衍生因子1的表达增高，与疾病严重程度呈正相关，可能参与了腰椎间盘退行性变的发病机制。

关键词: 组织构建, 组织构建与生物活性因子, 腰椎间盘退行性变, 基质细胞衍生因子1, 表达, 炎症, 趋化因子, Schneiderman分级, 细胞因子, 酶联免疫吸附, 细胞外基质, 基质金属蛋白酶

Abstract:

BACKGROUND: Exact pathogenesis of lumbar disc degeneration is not very clear. Inflammation is involved in the mechanism of lumbar disc degeneration. Stromal cell derived factor-1, which belongs to chemokine family, is related to inflammation.
OBJECTIVE: To determine the expression of stromal cell derived factor-1 in the intervertebral disk of patients with lumbar disc degeneration, and then to analyze the association of the expression levels of stromal cell derived factor-1 with the severity of lumbar disc degeneration.
METHODS: Eighty-four patients with lumbar disc degeneration (lumbar disc degeneration group) and 28 patients with disc burst fractures (control group) were enrolled in this study. The intervertebral disk tissues were collected after surgery. The protein level of stromal cell derived factor-1 was determined by enzyme linked immunosorbent assay. The grade of disc degeneration was determined according to Schneiderman’s classification. Association between the levels of stromal cell derived factor-1 and the Schneiderman’s grade was analyzed.
RESULTS AND CONCLUSION: The stromal cell derived factor-1 levels in the intervertebral disk were significantly elevated in the lumbar disc degeneration group compared with the control group (P < 0.01). The levels of stromal cell derived factor-1 were significantly increased in lumbar disc degeneration patients with grade 4 compared with those with grades 2 and 3. Furthermore, lumbar disc degeneration patients with grade 3 showed significantly elevated stromal cell derived factor-1 levels compared with those with grade 2. Spearman correlation analysis indicated that stromal cell derived factor-1 levels were positively correlated with Schneiderman’s grades (r=0.412, P < 0.01). These findings indicate that the levels of stromal cell derived factor-1 are significantly elevated in the intervertebral disk of patients with lumbar disc degeneration, and positively associated with the severity of lumbar disc degeneration. Stromal cell derived factor-1 may be involved in the mechanism of lumbar disc degeneration.

Key words: tissue construction, tissue construction and bioactive factors, lumbar disc degeneration, stromal cell derived factor-1, expression, inflammation, chemokines, Schneiderman’s grades, cytokines, enzyme-linked immunosorbnent assay, extracellular matrix, matrix metalloproteinase

中图分类号:

R318

刘钢，马信龙，邓树才，陈思. 退行性变腰椎间盘中基质细胞衍生因子1的表达及意义[J]. 中国组织工程研究, 2013, 17(24): 4488-4494.

Liu Gang, Ma Xin-long, Deng Shu-cai, Chen Si. Expression and significance of stromal cell derived factor-1 in the intervertebral disk after lumbar disc degeneration[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(24): 4488-4494.

图/表（结果） 2

参考文献

1. Jaerve A, Bosse F, Müller HW. SDF-1/CXCL12: its role in spinal cord injury. Int J Biochem Cell Biol. 2012;44(3):452-456. http://www.ncbi.nlm.nih.gov/pubmed?term=SDF-1%2FCXCL12%3A%20its%20role%20in%20spinal%20cord%20injury.%20Int%20J%20Biochem%20Cell%20Biol

2. Kohmo S, Kijima T, Mori M, et al. CXCL12 as a biological marker for the diagnosis of tuberculous pleurisy. Tuberculosis (Edinb). 2012;92(3):248-252. http://www.ncbi.nlm.nih.gov/pubmed/22297026

3. Wendel C, Hemping-Bovenkerk A, Krasnyanska J, et al. CXCR4/CXCL12 participate in extravasation of metastasizing breast cancer cells within the liver in a rat model. PLoS One. 2012;7(1):e30046.http://www.ncbi.nlm.nih.gov/pubmed?term=CXCR4%2FCXCL12%20participate%20in%20extravasation%20of%20metastasizing%20breast%20cancer%20cells%20within%20the%20liver%20in%20a%20rat%20model

4. Petruzziello-Pellegrini TN, Yuen DA, Page AV, et al. The CXCR4/CXCR7/SDF-1 pathway contributes to the pathogenesis of Shiga toxin-associated hemolytic uremic syndrome in humans and mice. J Clin Invest. 2012;122(2):759-776. http://www.ncbi.nlm.nih.gov/pubmed?term=The%20CXCR4%2FCXCR7%2FSDF-1%20pathway%20contributes%20to%20the%20pathogenesis%20of%20Shiga%20toxin-associated%20hemolytic%20uremic%20syndrome%20in%20humans%20and%20mice

5. Georgiou KR, Scherer MA, King TJ, et al. Deregulation of the CXCL12/CXCR4 axis in methotrexate chemotherapy-induced damage and recovery of the bone marrow microenvironment. Int J Exp Pathol. 2012;93(2):104-114. http://www.ncbi.nlm.nih.gov/pubmed?term=Deregulation%20of%20the%20CXCL12%2FCXCR4%20axis%20in%20methotrexate%20chemotherapy-induced%20damage%20and%20recovery%20of%20the%20bone%20marrow%20microenvironment

6. Ramos EA, Grochoski M, Braun-Prado K, et al. Epigenetic changes of CXCR4 and its ligand CXCL12 as prognostic factors for sporadic breast cancer. PLoS One. 2011;6(12):e29461.http://www.ncbi.nlm.nih.gov/pubmed?term=Epigenetic%20changes%20of%20CXCR4%20and%20its%20ligand%20CXCL12%20as%20prognostic%20factors%20for%20sporadic%20breast%20cancer

7. Wang Y, Huang J, Li Y, et al. Roles of chemokine CXCL12 and its receptors in ischemic stroke. Curr Drug Targets. 2012;13(2):166-172.http://www.ncbi.nlm.nih.gov/pubmed/22204316

8. Gordon CT, Wade C, Brinas I, et al. CXCL14 expression during chick embryonic development. Int J Dev Biol. 2011;55(3):335-340.http://www.ncbi.nlm.nih.gov/pubmed/21710440

9. Badr G, Mohany M, Metwalli A. Effects of undenatured whey protein supplementation on CXCL12- and CCL21-mediated B and T cell chemotaxis in diabetic mice. Lipids Health Dis. 2011;10:203.http://www.ncbi.nlm.nih.gov/pubmed?term=ffects%20of%20undenatured%20whey%20protein%20supplementation%20on%20CXCL12-%20and%20CCL21-mediated%20B%20and%20T%20cell%20chemotaxis%20in%20diabetic%20mice

10. Schmid MC, Avraamides CJ, Foubert P, et al. Combined blockade of integrin-α4β1 plus cytokines SDF-1α or IL-1β potently inhibits tumor inflammation and growth. Cancer Res. 2011;71(22):6965-6975.http://www.ncbi.nlm.nih.gov/pubmed?term=Combined%20blockade%20of%20integrin-%CE%B14%CE%B21%20plus%20cytokines%20SDF-1%CE%B1%20or%20IL-1%CE%B2%20potently%20inhibits%20tumor%20inflammation%20and%20growth

11. Kremer KN, Kumar A, Hedin KE. G alpha i2 and ZAP-70 mediate RasGRP1 membrane localization and activation of SDF-1-induced T cell functions. J Immunol. 2011;187(6):3177-3185.http://www.ncbi.nlm.nih.gov/pubmed?term=G%20alpha%20i2%20and%20ZAP-70%20mediate%20RasGRP1%20membrane%20localization%20and%20activation%20of%20SDF-1-induced%20T%20cell%20functions

12. Sisay Z, Berhe N, Petros B, et al. Serum chemokine profiles in visceral leishmaniasis, HIV and HIV/ visceral leishmaniasis co-infected Ethiopian patients. Ethiop Med J. 2011;49(3):179-186.http://www.ncbi.nlm.nih.gov/pubmed?term=Serum%20chemokine%20profiles%20in%20visceral%20leishmaniasis%2C%20HIV%20and%20HIV%2F%20visceral%20leishmaniasis%20co-infected%20Ethiopian%20patients

13. Popple A, Durrant LG, Spendlove I, et al. The chemokine, CXCL12, is an independent predictor of poor survival in ovarian cancer. Br J Cancer. 2012;106(7):1306-1313. http://www.ncbi.nlm.nih.gov/pubmed?term=The%20chemokine%2C%20CXCL12%2C%20is%20an%20independent%20predictor%20of%20poor%20survival%20in%20ovarian%20cancer

14. Weiler C, Lopez-Ramos M, Mayer HM, et al. Histological analysis of surgical lumbar intervertebral disc tissue provides evidence for an association between disc degeneration and increased body mass index. BMC Res Notes. 2011;4:497.http://www.ncbi.nlm.nih.gov/pubmed?term=Histological%20analysis%20of%20surgical%20lumbar%20intervertebral%20disc%20tissue%20provides%20evidence%20for%20an%20association%20between%20disc%20degeneration%20and%20increased%20body%20mass%20index

15. Liu LT, Huang B, Li CQ, et al. Characteristics of stem cells derived from the degenerated human intervertebral disc cartilage endplate. PLoS One. 2011;6(10):e26285. http://www.ncbi.nlm.nih.gov/pubmed?term=Characteristics%20of%20stem%20cells%20derived%20from%20the%20degenerated%20human%20intervertebral%20disc%20cartilage%20endplate

16. Choi KC, Kim JS, Kang BU, et al. Changes in back pain after percutaneous endoscopic lumbar discectomy and annuloplasty for lumbar disc herniation: a prospective study. Pain Med. 2011;12(11):1615-1621. http://www.ncbi.nlm.nih.gov/pubmed/?term=Changes+in+back+pain+after+percutaneous+endoscopic+lumbar+discectomy+and+annuloplasty+for+lumbar+disc+herniation%3A+a+prospective+study.

17. Inoue N, Espinoza Orías AA. Biomechanics of intervertebral disk degeneration. Orthop Clin North Am. 2011;42(4):487-499.http://www.ncbi.nlm.nih.gov/pubmed/21944586

18. Podichetty VK. The aging spine: the role of inflammatory mediators in intervertebral disc degeneration. Cell Mol Biol (Noisy-le-grand). 2007;53(5):4-18.http://www.ncbi.nlm.nih.gov/pubmed/17543240

19. Schneiderman G, Flannigan B, Kingston S, et al. Magnetic resonance imaging in the diagnosis of disc degeneration: correlation with discography. Spine (Phila Pa 1976). 1987;12(3):276-281.http://www.ncbi.nlm.nih.gov/pubmed/2954224

20. Tong Y, Xu W, Han H, et al. Tanshinone IIA increases recruitment of bone marrow mesenchymal stem cells to infarct region via up-regulating stromal cell-derived factor-1/CXC chemokine receptor 4 axis in a myocardial ischemia model. Phytomedicine. 2011;18(6):443-450.http://www.ncbi.nlm.nih.gov/pubmed?term=Tanshinone%20IIA%20increases%20recruitment%20of%20bone%20marrow%20mesenchymal%20stem%20cells%20to%20infarct%20region%20via%20up-regulating%20stromal%20cell-derived%20factor-1%2FCXC%20chemokine%20receptor%204%20axis%20in%20a%20myocardial%20ischemia%20model

21. Antonsson B, De Lys P, Dechavanne V, et al. In vivo processing of CXCL12α/SDF-1α after intravenous and subcutaneous administration to mice. Proteomics. 2010;10(24):4342-4351.http://www.ncbi.nlm.nih.gov/pubmed?term=In%20vivo%20processing%20of%20CXCL12%CE%B1%2FSDF-1%CE%B1%20after%20intravenous%20and%20subcutaneous%20administration%20to%20mice

22. Sarkar A, Tatlidede S, Scherer SS, et al. Combination of stromal cell-derived factor-1 and collagen-glycosaminoglycan scaffold delays contraction and accelerates reepithelialization of dermal wounds in wild-type mice. Wound Repair Regen. 2011;19(1):71-79.http://www.ncbi.nlm.nih.gov/pubmed?term=ICombination%20of%20stromal%20cell-derived%20factor-1%20and%20collagen-glycosaminoglycan%20scaffold%20delays%20contraction%20and%20accelerates%20reepithelialization%20of%20dermal%20wounds%20in%20wild-type%20mice

23. Fiorina P, Jurewicz M, Vergani A, et al. Targeting the CXCR4-CXCL12 axis mobilizes autologous hematopoietic stem cells and prolongs islet allograft survival via programmed death ligand 1. J Immunol. 2011;186(1):121-131. http://www.ncbi.nlm.nih.gov/pubmed?term=Targeting%20the%20CXCR4-CXCL12%20axis%20mobilizes%20autologous%20hematopoietic%20stem%20cells%20and%20prolongs%20islet%20allograft%20survival%20via%20programmed%20death%20ligand%201

24. Benamar K, Palma J, Cowan A, et al. Analgesic efficacy of buprenorphine in the presence of high levels of SDF-1α/CXCL12 in the brain. Drug Alcohol Depend. 2011;114(2-3):246-248.http://www.ncbi.nlm.nih.gov/pubmed?term=Analgesic%20efficacy%20of%20buprenorphine%20in%20the%20presence%20of%20high%20levels%20of%20SDF-1%CE%B1%2FCXCL12%20in%20the%20brain

25. Jung Y, Shiozawa Y, Wang J, et al. Annexin-2 is a regulator of stromal cell-derived factor-1/CXCL12 function in the hematopoietic stem cell endosteal niche. Exp Hematol. 2011;39(2):151-166.http://www.ncbi.nlm.nih.gov/pubmed?term=Annexin-2%20is%20a%20regulator%20of%20stromal%20cell-derived%20factor-1%2FCXCL12%20function%20in%20the%20hematopoietic%20stem%20cell%20endosteal%20niche

26. Rhodes LV, Antoon JW, Muir SE, et al. Effects of human mesenchymal stem cells on ER-positive human breast carcinoma cells mediated through ER-SDF-1/CXCR4 crosstalk. Mol Cancer. 2010;9:295.http://www.ncbi.nlm.nih.gov/pubmed?term=Effects%20of%20human%20mesenchymal%20stem%20cells%20on%20ER-positive%20human%20breast%20carcinoma%20cells%20mediated%20through%20ER-SDF-1%2FCXCR4%20crosstalk

27. Moll NM, Ransohoff RM. CXCL12 and CXCR4 in bone marrow physiology. Expert Rev Hematol. 2010;3(3):315-322.http://www.ncbi.nlm.nih.gov/pubmed/21082982

28. Rhodes LV, Short SP, Neel NF, et al. Cytokine receptor CXCR4 mediates estrogen-independent tumorigenesis, metastasis, and resistance to endocrine therapy in human breast cancer. Cancer Res. 2011;71(2):603-613. http://www.ncbi.nlm.nih.gov/pubmed?term=Cytokine%20receptor%20CXCR4%20mediates%20estrogen-independent%20tumorigenesis%2C%20metastasis%2C%20and%20resistance%20to%20endocrine%20therapy%20in%20human%20breast%20cancer

29. Ma L, Qiao H, He C, et al. Modulating the interaction of CXCR4 and CXCL12 by low-molecular-weight heparin inhibits hepatic metastasis of colon cancer. Invest New Drugs. 2012;30(2):508-517. http://www.ncbi.nlm.nih.gov/pubmed?term=Modulating%20the%20interaction%20of%20CXCR4%20and%20CXCL12%20by%20low-molecular-weight%20heparin%20inhibits%20hepatic%20metastasis%20of%20colon%20cancer

30. Patrussi L, Baldari CT. The CXCL12/CXCR4 axis as a therapeutic target in cancer and HIV-1 infection. Curr Med Chem. 2011;18(4):497-512.http://www.ncbi.nlm.nih.gov/pubmed/21143114

31. Jia CQ, Zhao JG, Zhang SF, et al. Stromal cell-derived factor-1 and vascular endothelial growth factor may play an important role in the process of neovascularization of herniated intervertebral discs. J Int Med Res. 2009;37(1):136-144.http://www.ncbi.nlm.nih.gov/pubmed?term=Stromal%20cell-derived%20factor-1%20and%20vascular%20endothelial%20growth%20factor%20may%20play%20an%20important%20role%20in%20the%20process%20of%20neovascularization%20of%20herniated%20intervertebral%20discs

32. Samartzis D, Cheung KM. Lumbar intervertebral disk degeneration. Orthop Clin North Am. 2011 Oct;42(4):xi-xii.http://www.ncbi.nlm.nih.gov/pubmed/21944595

33. Omair A, Lie BA, Reikeras O, et al. An Association Study of Interleukin 18 Receptor Genes (IL18R1 and IL18RAP) in Lumbar Disc Degeneration. Open Orthop J. 2012;6:164-171.http://www.ncbi.nlm.nih.gov/pubmed?term=An%20Association%20Study%20of%20Interleukin%2018%20Receptor%20Genes%20(IL18R1%20and%20IL18RAP)%20in%20Lumbar%20Disc%20Degeneration

34. Podichetty VK. The aging spine: the role of inflammatory mediators in intervertebral disc degeneration. Cell Mol Biol (Noisy-le-grand). 2007;53(5):4-18.http://www.ncbi.nlm.nih.gov/pubmed/17543240

35. Valdes AM, Hassett G, Hart DJ, et al. Radiographic progression of lumbar spine disc degeneration is influenced by variation at inflammatory genes: a candidate SNP association study in the Chingford cohort. Spine (Phila Pa 1976). 2005;30(21):2445-2451.http://www.ncbi.nlm.nih.gov/pubmed?term=Radiographic%20progression%20of%20lumbar%20spine%20disc%20degeneration%20is%20influenced%20by%20variation%20at%20inflammatory%20genes%3A%20a%20candidate%20SNP%20association%20study%20in%20the%20Chingford%20cohort

36. Bachmeier BE, Nerlich AG, Weiler C, et al. Analysis of tissue distribution of TNF-alpha, TNF-alpha-receptors, and the activating TNF-alpha-converting enzyme suggests activation of the TNF-alpha system in the aging intervertebral disc. Ann N Y Acad Sci. 2007;1096:44-54.http://www.ncbi.nlm.nih.gov/pubmed?term=Analysis%20of%20tissue%20distribution%20of%20TNF-alpha%2C%20TNF-alpha-receptors%2C%20and%20the%20activating%20TNF-alpha-converting%20enzyme%20suggests%20activation%20of%20the%20TNF-alpha%20system%20in%20the%20aging%20intervertebral%20disc

37. Studer RK, Vo N, Sowa G, et al. Human nucleus pulposus cells react to IL-6: independent actions and amplification of response to IL-1 and TNF-α. Spine (Phila Pa 1976). 2011;36(8):593-599.http://www.ncbi.nlm.nih.gov/pubmed?term=Human%20nucleus%20pulposus%20cells%20react%20to%20IL-6%3A%20independent%20actions%20and%20amplification%20of%20response%20to%20IL-1%20and%20TNF-%CE%B1

38. Jimbo K, Park JS, Yokosuka K, et al. Positive feedback loop of interleukin-1beta upregulating production of inflammatory mediators in human intervertebral disc cells in vitro. J Neurosurg Spine. 2005;2(5):589-595.http://www.ncbi.nlm.nih.gov/pubmed?term=Positive%20feedback%20loop%20of%20interleukin-1beta%20upregulating%20production%20of%20inflammatory%20mediators%20in%20human%20intervertebral%20disc%20cells%20in%20vitro

39. Igarashi A, Kikuchi S, Konno S, et al. Inflammatory cytokines released from the facet joint tissue in degenerative lumbar spinal disorders. Spine (Phila Pa 1976). 2004;29(19):2091-2095.http://www.ncbi.nlm.nih.gov/pubmed/15454697

40. O'Neill CW, Liu JJ, Leibenberg E, et al. Percutaneous plasma decompression alters cytokine expression in injured porcine intervertebral discs. Spine J. 2004;4(1):88-98.http://www.ncbi.nlm.nih.gov/pubmed?term=Percutaneous%20plasma%20decompression%20alters%20cytokine%20expression%20in%20injured%20porcine%20intervertebral%20discs

41. Hamamoto H, Miyamoto H, Doita M, et al. Capability of nondegenerated and degenerated discs in producing inflammatory agents with or without macrophage interaction. Spine (Phila Pa 1976). 2012;37(3):161-167.http://www.ncbi.nlm.nih.gov/pubmed/21494199

42. Darisipudi MN, Kulkarni OP, Sayyed SG, et al. Dual blockade of the homeostatic chemokine CXCL12 and the proinflammatory chemokine CCL2 has additive protective effects on diabetic kidney disease. Am J Pathol. 2011 Jul;179(1):116-124.http://www.ncbi.nlm.nih.gov/pubmed?term=Dual%20blockade%20of%20the%20homeostatic%20chemokine%20CXCL12%20and%20the%20proinflammatory%20chemokine%20CCL2%20has%20additive%20protective%20effects%20on%20diabetic%20kidney%20disease

43. Boldajipour B, Doitsidou M, Tarbashevich K, et al. Cxcl12 evolution--subfunctionalization of a ligand through altered interaction with the chemokine receptor. Development. 2011;138(14):2909-2914.http://www.ncbi.nlm.nih.gov/pubmed?term=Cxcl12%20evolution--subfunctionalization%20of%20a%20ligand%20through%20altered%20interaction%20with%20the%20chemokine%20receptor

44. Ping YF, Yao XH, Jiang JY, et al. The chemokine CXCL12 and its receptor CXCR4 promote glioma stem cell-mediated VEGF production and tumour angiogenesis via PI3K/AKT signalling. J Pathol. 2011;224(3):344-354. http://www.ncbi.nlm.nih.gov/pubmed?term=The%20chemokine%20CXCL12%20and%20its%20receptor%20CXCR4%20promote%20glioma%20stem%20cell-mediated%20VEGF%20production%20and%20tumour%20angiogenesis%20via%20PI3K%2FAKT%20signalling

45. Zhang Y, Tian L, Zheng Y, et al. C-terminal peptides of chemokine-like factor 1 signal through chemokine receptor CCR4 to cross-desensitize the CXCR4. Biochem Biophys Res Commun. 2011;409(2):356-361. http://www.ncbi.nlm.nih.gov/pubmed?term=C-terminal%20peptides%20of%20chemokine-like%20factor%201%20signal%20through%20chemokine%20receptor%20CCR4%20to%20cross-desensitize%20the%20CXCR4

46. Delano MJ, Kelly-Scumpia KM, Thayer TC, et al. Neutrophil mobilization from the bone marrow during polymicrobial sepsis is dependent on CXCL12 signaling. J Immunol. 2011;187(2):911-918.http://www.ncbi.nlm.nih.gov/pubmed?term=Neutrophil%20mobilization%20from%20the%20bone%20marrow%20during%20polymicrobial%20sepsis%20is%20dependent%20on%20CXCL12%20signaling

47. Hashikawa K, Niino D, Yasumoto S, et al. Clinicopathological features and prognostic significance of CXCL12 in blastic plasmacytoid dendritic cell neoplasm. J Am Acad Dermatol. 2012;66(2):278-291. http://www.ncbi.nlm.nih.gov/pubmed?term=Clinicopathological%20features%20and%20prognostic%20significance%20of%20CXCL12%20in%20blastic%20plasmacytoid%20dendritic%20cell%20neoplasm

48. English K. Intervertebral disc repair: mesenchymal stem cells to the rescue? Transplantation. 2011;92(7):733-734.http://www.ncbi.nlm.nih.gov/pubmed?term=Intervertebral%20disc%20repair%3A%20mesenchymal%20stem%20cells%20to%20the%20rescue%3F

49. Kelempisioti A, Eskola PJ, Okuloff A, et al. Genetic susceptibility of intervertebral disc degeneration among young Finnish adults. BMC Med Genet. 2011;12:153.http://www.ncbi.nlm.nih.gov/pubmed/22107760

50. Jünger S, Gantenbein-Ritter B, Lezuo P, et al. Effect of limited nutrition on in situ intervertebral disc cells under simulated-physiological loading. Spine (Phila Pa 1976). 2009;34(12):1264-1271.http://www.ncbi.nlm.nih.gov/pubmed?term=Effect%20of%20limited%20nutrition%20on%20in%20situ%20intervertebral%20disc%20cells%20under%20simulated-physiological%20loading

51. Zigouris A, Alexiou GA, Batistatou A, et al. The role of matrix metalloproteinase 9 in intervertebral disc degeneration. J Clin Neurosci. 2011;18(10):1424-1425.http://www.ncbi.nlm.nih.gov/pubmed/21763143

52. Yurube T, Nishida K, Suzuki T, et al. Matrix metalloproteinase (MMP)-3 gene up-regulation in a rat tail compression loading-induced disc degeneration model. J Orthop Res. 2010;28(8):1026-1032.http://www.ncbi.nlm.nih.gov/pubmed?term=Matrix%20metalloproteinase%20(MMP)-3%20gene%20up-regulation%20in%20a%20rat%20tail%20compression%20loading-induced%20disc%20degeneration%20model

53. Zigouris A, Alexiou GA, Batistatou A, et al. The role of matrix metalloproteinase 9 in intervertebral disc degeneration. J Clin Neurosci. 2011;18(10):1424-1425. http://www.ncbi.nlm.nih.gov/pubmed/21763143

54. Zigouris A, Batistatou A, Alexiou GA, et al. Correlation of matrix metalloproteinases-1 and -3 with patient age and grade of lumbar disc herniation. J Neurosurg Spine. 2011;14(2):268-272.http://www.ncbi.nlm.nih.gov/pubmed?term=Correlation%20of%20matrix%20metalloproteinases-1%20and%20-3%20with%20patient%20age%20and%20grade%20of%20lumbar%20disc%20herniation

55. Kanbe K, Takagishi K, Chen Q. Stimulation of matrix metalloprotease 3 release from human chondrocytes by the interaction of stromal cell-derived factor 1 and CXC chemokine receptor 4. Arthritis Rheum. 2002;46(1):130-137.http://www.ncbi.nlm.nih.gov/pubmed?term=Stimulation%20of%20matrix%20metalloprotease%203%20release%20from%20human%20chondrocytes%20by%20the%20interaction%20of%20stromal%20cell-derived%20factor%201%20and%20CXC%20chemokine%20receptor%204

56. Kanbe K, Takemura T, Takeuchi K, et al. Synovectomy reduces stromal-cell-derived factor-1 (SDF-1) which is involved in the destruction of cartilage in osteoarthritis and rheumatoid arthritis. J Bone Joint Surg Br. 2004;86(2):296-300.http://www.ncbi.nlm.nih.gov/pubmed?term=ynovectomy%20reduces%20stromal-cell-derived%20factor-1%20(SDF-1)%20which%20is%20involved%20in%20the%20destruction%20of%20cartilage%20in%20osteoarthritis%20and%20rheumatoid%20arthritis

57. Kanbe K, Takagishi K, Chen Q. Stimulation of matrix metalloprotease 3 release from human chondrocytes by the interaction of stromal cell-derived factor 1 and CXC chemokine receptor 4. Arthritis Rheum. 2002;46(1):130-137.http://www.ncbi.nlm.nih.gov/pubmed?term=Stimulation%20of%20matrix%20metalloprotease%203%20release%20from%20human%20chondrocytes%20by%20the%20interaction%20of%20stromal%20cell-derived%20factor%201%20and%20CXC%20chemokine%20receptor%204

引言

材料方法

设计：对比观察试验。

时间和地点：于2011年6月至2012年4月在天津医院完成。

对象：选取2011年6月至2011年4月在天津医院接受手术治疗的腰椎间盘退行性变患者84例作为病例组，另外选取同时期腰椎爆裂性骨折患者28例作为对照组。

病例组：84例患者中，男54例，女30例；年龄42-58岁，平均47.2岁。

诊断标准：符合腰椎间盘退行性变的诊断标准，并经MRI证实。

纳入标准：①经MRI证实存在腰椎间盘退行性变的患者。②接受手术治疗的患者。③对治疗及试验方案知情同意，且得到医院伦理委员会批准者。

排除标准：合并有腰椎管狭窄者。

对照组：28例患者中，男18例，女10例；年龄32-56岁，平均46.8岁。

诊断标准：符合腰椎爆裂性骨折的诊断标准，并经MRI证实。

纳入标准：①伤前无慢性腰背痛史，MRI示无椎间盘退变征象，术后组织学检查椎间盘组织无明显退变者。②对治疗及试验方案知情同意，且得到医院伦理委员会批准者。

排除标准：①恶性肿瘤患者。②急、慢性炎症性疾病患者。③自身免疫性疾病或结缔组织疾病患者。

方法：

椎间盘标本的制备：收集2组患者手术摘除的退变椎间盘组织，置于液氮罐中保存和运输，过夜后于次日清晨转移至-80 ℃保存。

酶联免疫吸附方法检测基质细胞衍生因子1表达水平：①将椎间盘组织标本用无菌生理盐水反复冲洗，然后研磨成匀浆，加入磷酸缓冲液至5 mL。3 000 r/min离心5 min后，留取上清液，用于检测基质细胞衍生因子1水平。②配液：用试剂盒自带的稀释液将样品上清液稀释10倍。③加样：酶标板分别标记标准品孔、待测样本孔和空白对照孔，记录各孔位置，取标准品50 μL加入到标准品孔中，将稀释后的10 μL待测样本加入到待测样本孔中，空白对照孔不加。④孵育：将酶标板置于37 ℃恒温箱孵育30 min。⑤洗板：弃去各孔中的液体，吸水纸上拍干，然后在每孔中加满洗涤液，静置3 min后，甩去洗涤液，再次在吸水纸上拍干，重复4次。⑥加酶标工作液：每孔加入酶标工作液50 μL，空白对照孔不加。⑦孵育：将酶标板置于37 ℃恒温箱孵育30 min。⑧洗板：弃去各孔中的液体，吸水纸上拍干，然后在每孔中加满洗涤液，静置3 min后，甩去洗涤液，再次在吸水纸上拍干，重复4次。⑨显色：每孔先加入显色剂A液50 μL，再加入显色剂B液50 μL，平混匀后于室温下避光显色15 min。⑩终止：每孔加入终止液50 μL，终止反应。?测定：以空白孔调零，在终止后15 min内，用450 nm波长测量各孔的吸光度。?计算：根据标准品的浓度及对应吸光度，计算出对应的样品浓度。

Schneiderman分级：84例腰椎间盘退行性变患者行MRI检查，MRI扫描时为脊柱协同线圈，T2加权像时TE为120 ms，TR为3 500 ms，层厚/层距为4.0/0.4 mm。椎间盘退变程度根据MRI的T2加权像中椎间盘信号强度变化进行判断，由放射科及骨科医师各1名根据Schneiderman分级标准分为4级^[18]：1级，椎间盘信号正常；2级，椎间盘信号轻度减低；3级，椎间盘信号中度减低；4级，椎间盘信号完全消失。病例组中2级16例，3级46例，4级22例。

主要观察指标：腰椎间盘退行性变患者椎间盘组织中基质细胞衍生因子1的表达水平及其与Schneiderman分级的关系。

统计学分析：刘钢应用SPSS 13.0统计软件进行统计学处理，所有数据均用x(_)±s表示，采用两独立样本t 检验或方差分析比较组间的差异，以Spearman分析方法分析基质细胞衍生因子1与Schneiderman分级的相关性，P < 0.05为差异有显著性意义。

讨论

文章结果显示，腰椎间盘退行性变患者的腰椎间盘组织中基质细胞衍生因子1水平明显升高，基质细胞衍生因子1水平与Schneiderman分级显著相关，该研究为国内首次涉及基质细胞衍生因子1在退变椎间盘组织中表达的研究。

基质细胞衍生因子1是一类新近发现的趋化因子家族成员，也称为前B细胞增长刺激因子或者CXCL12。基质细胞衍生因子1是对骨髓细胞趋化效应最强的趋化因子，趋化活性较其他的趋化因子高^[20]，基质细胞衍生因子1在许多方面与其他的趋化因子不同：①基质细胞衍生因子1有高度的保守性，小鼠和人的基质细胞衍生因子1有99%的同源性性，仅存在1个氨基酸的差异^[21]。②通常一种趋化因子可以结合多种受体，一种受体也可结合多种趋化因子，但迄今发现只有基质细胞衍生因子1与CXCR4是一对一的结合关系^[22]。③多数CXC化学因子都位于人4q12-q21染色体上，而基质细胞衍生因子1则位于人10q11.1染色体上^[23]。④基质细胞衍生因子1主要由低氧的情况下诱导基质细胞持续分泌而产生，而其他趋化因子则一般都是在炎症状态下，由炎症细胞表达和产生^[24]。⑤每个趋化因子家族都共有高度相似的序列和相似的生物学特异性光谱，但基质细胞衍生因子1α的结构序列与其他CXC和CC趋化因子有很大差异，序列相似性平均仅为20%-25%^[25]。

CXCR4为G蛋白偶联受体，其结构主要由7个疏水的跨膜区、细胞外的氨基端、胞质中的羧基端及细胞内、外的3个环组成^[26]。CXCR4是目前已知基质细胞衍生因子1惟一的受体，基质细胞衍生因子1与CXCR4的结构和相互作用是发挥其病理、生理功能的基础^[27]。基质细胞衍生因子1的N-端氨基酸残基是与CXCR4相互作用的关键区域，如N-端氨基酸残基缺失、突变则不能与其受体结合，C-端能极大地增强N-端肽段的信号转导^[28]。基质细胞衍生因子1/CXCR4是一个主要的化学趋化因子/受体轴，这个轴在许多生理过程，包括造血、心脏形成、新生血管生成、神经发育和免疫细胞的运输中起关键的作用^[29]，在肿瘤转移、人类免疫缺陷综合征、炎症过程以及组织缺血等病理状态下也起了一定的作用^[30]。

文章结果表明，腰椎间盘组织中基质细胞衍生因子1水平与腰椎间盘退行性变的发病和严重程度相关，基质细胞衍生因子1可以作为预测腰椎间盘退行性变发病风险和疾病进展的一项生物学指标。同时，腰椎间盘退行性变患者椎间盘组织中升高的基质细胞衍生因子1水平也提示，基质细胞衍生因子1可能参与了腰椎间盘退行性变的病理生理机制。近来的一项研究显示，椎间盘突出患者的椎间盘组织中基质细胞衍生因子1的表达明显高于对照组，并且基质细胞衍生因子1与血管内皮生长因子的表达水平显著相关，提示基质细胞衍生因子1与腰椎间盘退行性变的发病有关，并有可能通过与血管内皮生长因子相互作用，促进椎间盘组织中新生血管生成，最终导致椎间盘突出^[31]。

椎间盘退行性变不仅与椎间盘的生物力学改变等机械性因素有关^[32]，炎性细胞因子也参与了腰椎间盘退行性变的发病过程^[33-34]。细胞因子是一类由多种细胞产生并分泌到细胞外的小分子糖蛋白，包括肿瘤坏死因子α、白细胞介素6等，具有多种生物学功能，可介导炎症反应，在退行性骨关节病中发挥重要的作用^[35-41]。基质细胞衍生因子1，又称为CXCL12，属于趋化因子家族，具有趋化作用，能够趋化淋巴细胞、单核细胞和树突状细胞等各种炎性细胞^[42-47]。因而，基质细胞衍生因子1有可能通过趋化炎症细胞进入椎间盘组织，并促使炎症细胞释放各种炎症细胞因子，放大炎症反应，加重椎间盘组织损伤，最终导致腰椎间盘退行性变的发生。

近年来随着分子生物学及免疫学的深入研究，普遍认为椎间盘退变主要表现在细胞和细胞外基质成分的变化，而后者是椎间髓质力学特性丧失的直接原因^[48]。正常椎间盘基质的合成代谢和分解代谢之间存在着动态平衡^[49]，发生退变时，常有基质分解代谢的增强同时伴有合成代谢的减弱，二者之间平衡被打破^[50]。基质合成与降解的失衡将导致基质成分的紊乱，在此调节过程中基质金属蛋白酶和金属蛋白酶组织抑制剂2个酶系统发挥着重要作用^[51]。

正常椎间盘中基质金属蛋白酶都是以无活性的酶原形式分泌，然后在除去细胞间质内N-端结构域后才具有蛋白水解酶活性，基质金属蛋白酶可相互作用或与其他蛋白酶发生激活级联反应^[52]。体内的许多激素、细胞因子、蛋白水解酶、自由基等都可以促进基质金属蛋白酶的激活^[53]。椎间盘细胞外基质的主要成分是Ⅱ型胶原和蛋白多糖，退变椎间盘主要变化是Ⅱ型胶原和蛋白多糖的减少^[54]。研究表明，基质细胞衍生因子1可以促进各种基质金属蛋白酶，如基质金属蛋白酶3、基质金属蛋白酶9和基质金属蛋白酶13的表达和释放^[55-57]，提示基质细胞衍生因子1能够间接地通过调节基质金属蛋白酶的表达，间接地促进椎间盘细胞外基质的降解，导致腰椎间盘退行性变的病理生理变化。

研究尚存在几点不足之处。首先，本研究的样本量相对较小，还需要将来有大样本量的临床研究对该实验结论加以证实。另外，本文为横断面研究，希望能有前瞻性的临床研究来进一步研究基质细胞衍生因子1在腰椎间盘退行性变中的作用。

综上所述，腰椎间盘组织中基质细胞衍生因子1水平与腰椎间盘退行性变的发病和严重程度相关，椎间盘组织中的基质细胞衍生因子1表达水平可以作为预测腰椎间盘退行性变发病风险和疾病进展的一项生物学标记物。

文章快阅

延伸阅读

基质细胞衍生因子1是一类新近发现的趋化因子家族成员，也称为前B细胞增长刺激因子或者CXCL12。基质细胞衍生因子1是对骨髓细胞趋化效应最强的趋化因子，趋化活性较其他的趋化因子高，基质细胞衍生因子1在许多方面与其他的趋化因子不同：①基质细胞衍生因子1有高度的保守性，小鼠和人的基质细胞衍生因子1有99%的同源性性，仅存在1个氨基酸的差异。②通常一种趋化因子可以结合多种受体，一种受体也可结合多种趋化因子，但迄今发现只有基质细胞衍生因子1与CXCR4是一对一的结合关系。③多数CXC化学因子都位于人4q12-q21染色体上，而基质细胞衍生因子1则位于人10q11.1染色体上。④基质细胞衍生因子1主要由低氧的情况下诱导基质细胞持续分泌而产生，而其他趋化因子则一般都是在炎症状态下，由炎症细胞表达和产生。⑤每个趋化因子家族都共有高度相似的序列和相似的生物学特异性光谱，但基质细胞衍生因子1α的结构序列与其他CXC和CC趋化因子有很大差异，序列相似性平均仅为20％-25％。CXCR4为G蛋白偶联受体，其结构主要由7个疏水的跨膜区、细胞外的氨基端、胞质中的羧基端及细胞内、外的3个环组成。CXCR4是目前已知基质细胞衍生因子1惟一的受体，基质细胞衍生因子1与CXCR4的结构和相互作用是发挥其病理、生理功能的基础。基质细胞衍生因子1的N-端氨基酸残基是与CXCR4相互作用的关键区域，如N-端氨基酸残基缺失、突变则不能与其受体结合，C-端能极大地增强N-端肽段的信号转导。基质细胞衍生因子1/CXCR4是一个主要的化学趋化因子/受体轴，这个轴在许多生理过程，包括造血、心脏形成、新生血管生成、神经发育和免疫细胞的运输中起关键的作用，在肿瘤转移、人类免疫缺陷综合征、炎症过程以及组织缺血等病理状态下也起了一定的作用。

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