• 1 同濟(jì)大學(xué)醫(yī)學(xué)院附屬楊浦醫(yī)院骨科（上海，200090）;;
  • 2 英國(guó)牛津大學(xué)臨床學(xué)院Nuffield 骨科和Botnar 研究中心;;
  • 3 上海交通大學(xué)附屬第六人民醫(yī)院骨科;

目的 糖皮質(zhì)激素會(huì)破壞軟骨，通過觀察地塞米松對(duì)人骨關(guān)節(jié)炎軟骨細(xì)胞（human articular
chondrocytes，HACs）凋亡及Fas/FasL 基因表達(dá)的影響，探討其促進(jìn)HACs 凋亡的作用機(jī)制。 方法 取行人工膝關(guān)節(jié)
置換的膝骨關(guān)節(jié)炎患者自愿捐贈(zèng)軟骨組織，體外分離培養(yǎng)HACs，取第2 代細(xì)胞進(jìn)行實(shí)驗(yàn)。將HACs 分別置于含濃度為
0.125、1.25、12.5、25 及50 μg/mL 地塞米松培養(yǎng)液中，培養(yǎng)48 h 后采用MTT 法選擇地塞米松最佳工作濃度進(jìn)行后續(xù)實(shí)
驗(yàn)。將HACs 分別采用最佳工作濃度地塞米松（實(shí)驗(yàn)組）及不含地塞米松培養(yǎng)液（對(duì)照組）培養(yǎng)，0、24、48 h 后行TMRE/
Hoechst/Annexin V-FITC/7-AAD 四重染色檢測(cè)細(xì)胞凋亡，48 h 后行實(shí)時(shí)定量PCR 檢測(cè)Fas/FasL mRNA 表達(dá)，0、24、48 h
后行免疫組織化學(xué)染色檢測(cè)Fas/FasL 蛋白表達(dá)。 結(jié)果 25 μg/mL 濃度組細(xì)胞抑制率顯著高于50 μg/mL 濃度組，差異
有統(tǒng)計(jì)學(xué)意義（P  lt; 0.05）；與其他濃度組比較，差異亦有統(tǒng)計(jì)學(xué)意義（P  lt; 0.05）；選擇25 μg/mL 濃度作為后續(xù)實(shí)驗(yàn)工作濃
度。培養(yǎng)0、24、48 h，實(shí)驗(yàn)組HACs 凋亡細(xì)胞百分比分別為5.8% ± 0.3%、27.0% ± 2.6%、36.0% ± 3.1%，呈明顯時(shí)間依賴性（P  lt; 0.05）。培養(yǎng)48 h 后實(shí)驗(yàn)組及對(duì)照組Fas mRNA 相對(duì)表達(dá)量分別為（8.93 ± 1.12）× 10—3 及（3.31 ± 0.37）× 10—3，F(xiàn)asLmRNA 相對(duì)表達(dá)量分別為（5.92 ± 0.66）× 10—3 及（2.31 ± 0.35）× 10—3，兩組比較差異均有統(tǒng)計(jì)學(xué)意義（P  lt; 0.05）。隨培養(yǎng)時(shí)間延長(zhǎng)，實(shí)驗(yàn)組Fas 及FasL 蛋白表達(dá)逐漸增加，且各時(shí)間點(diǎn)均顯著高于對(duì)照組，差異均有統(tǒng)計(jì)學(xué)意義（P  lt; 0.05）。 結(jié)論 地塞米松可促進(jìn)HACs 細(xì)胞凋亡及上調(diào)凋亡基因Fas/FasL 表達(dá)，為進(jìn)一步探討Fas/FasL 信號(hào)通路在地塞米松促HACs 凋亡中的作用提供了實(shí)驗(yàn)依據(jù)。

引用本文： 涂意輝 ,薛華明,夏志道,蔡珉巍,劉曉東,馬童,張長(zhǎng)青. 不同濃度地塞米松對(duì)人骨關(guān)節(jié)炎軟骨細(xì)胞凋亡及Fas/FasL 基因表達(dá)的影響. 中國(guó)修復(fù)重建外科雜志, 2012, 26(5): 536-541. doi: 復(fù)制

1. Céleste C, Ionescu M, Robin Poole A, et al. Repeated intraarticular injections of triamcinolone acetonide alter cartilage matrix metabolism measured by biomarkers in synovial fluid. J Orthop Res, 2005, 23(3): 602-610.
2. Madsen SH, Andreassen KV, Christensen ST, et al. Glucocorticoids exert context-dependent effects on cells of the joint in vitro. Steroids, 2011, 76(13): 1474-1482.
3. Jaramillo MC, Briehl MM, Crapo JD, et al. Manganese porphyrin, MnTE-2-PyP (5+), acts as a pro-oxidant to potentiate glucocorticoid-induced apoptosis in lymphoma cells. Free Radic Biol Med, 2012. [Epub ahead of print].
4. Chung YJ, Lee JI, Chong S, et al. Anti-proliferative effect and action mechanism of dexamethasone in human medullary thyroid cancer cell line. Endocr Res, 2011, 36(4): 149-157.
5. Li WZ, Li WP, Huang DK, et al. Dexamethasone and Aβ25-35 accelerate learning and memory impairments due to elevate amyloid precursor protein expression and neuronal apoptosis in 12-month male rats. Behav Brain Res, 2012, 227(1): 142-149.
6. Zuloaga DG, Carbone DL, Hiroi R, et al. Dexamethasone induces apoptosis in the developing rat amygdala in an age-, region-, and sex-specific manner. Neuroscience, 2011, 199: 535-547.
7. Ortega-Ortega Y, García-Latorre EA, Lezama RA, et al. Heat shock protein 60 from Klebsiella pneumoniae protects mononuclear cells from apoptotic cell death induced by dexamethasone. Microb Pathog, 2011, 51(5): 352-359.
8. Silvestrini G, Ballanti P, Patacchioli FR, et al. Evaluation of apoptosis and the glucocorticoid receptor in the cartilage growth plate and metaphyseal bone cells of rats after high-dose treatment with corticosterone. Bone, 2000, 26(1): 33-42.
9. Hochberg MC, Altman RD, Brandt KD, et al. Guidelines for the medical management of osteoarthritis. Part II. Osteoarthritis of the knee. American College of Rheumatology Arthritis Rheum, 1995, 38(11): 1541-1546.
10. Dragoo JL, Danial CM, Braun HJ, et al. The chondrotoxicity of single-dose corticosteroids. Knee Surg Sports Traumatol Arthrosc, 2011, [Epub ahead of print].
11. Braun HJ, Wilcox-Fogel N, Kim HJ, et al. The effect of local anesthetic and corticosteroid combinations on chondrocyte viability. Knee Surg Sports Traumatol Arthrosc, 2011. [Epub ahead of print].
12. O’Driscoll SW. The healing and regeneration of articular cartilage. J Bone Joint Surg (Am), 1998, 80(12): 1795-1812.
13. Hossain MA, Park J, Choi SH, et al. Dexamethasone induces apoptosis in proliferative canine tendon cells and chondrocytes. Vet Comp Orthop Traumatol, 2008, 21(4): 337-342.
14. Chrysis D, Ritzen EM, Sävendahl L. Growth retardation induced by dexamethasone is associated with increased apoptosis of the growth plate chondrocytes. J Endocrinol, 2003, 176(3): 331-337.
15. Chrysis D, Zaman F, Chagin AS, et al. Dexamethasone induced apoptosis in proliferative chondrocytes through activation of caspases and suppression of the Akt-phosphatidyinositol 3’-kinase signaling pathway. Endocrinology, 2005, 146(3): 1391-1397.
16. Jia J, Yao W, Guan M, et al. Glucocorticoid dose determines osteocyte cell fate. FASEB J, 2011, 25(10): 3366-3376.
17. Lotz M, Hashimoto S, Kühn K. Mechanisms of chondrocyte apoptosis. Osteoarthritis Cartilage, 1999, 7(4): 389-391.
18. Kim HA, Blanco FJ. Cell death and apoptosis in osteoarthritic cartilage. Curr Drug Targets, 2007, 8(2): 333-345.
19. Lavrik IN. Regulation of death receptor-induced apoptosis induced via CD95/FAS and other death receptors. Mol Biol (Mosk), 2011, 45(1): 173-179.
20. 王玉, 孫黎光, 夏春輝. Caspase介導(dǎo)的Fas凋亡途徑. 世界華人消化雜志, 2006, 14(36): 3439-3442.
21. Naves MA, Pereira RM, Comodo AN, et al. Effect of dexamethasone on human osteoblasts in culture: involvement of β1 integrin and integrin-linked kinase. Cell Biol Int, 2011, 35(11): 1147-1151.
22. 肖娟, 曹誼林, 范華驊, 等. 體外培養(yǎng)軟骨細(xì)胞凋亡與caspase-3表達(dá)的實(shí)驗(yàn)研究. 中國(guó)修復(fù)重建外科雜志, 2003, 17(5): 418-421.
23. Wang HQ, Yu XD, Liu ZH, et al. Deregulated miR-155 promotes Fas-mediated apoptosis in human intervertebral disc degeneration by targeting FADD and caspase-3. J Pathol, 2011, 225(2): 232-242.
24. von Rossum A, Krall R, Escalante NK, et al. Inflammatory cytokines determine the susceptibility of human CD8 T cells to Fas-mediated activation-induced cell death through modulation of FasL and c-FLIP (S) expression. J Biol Chem, 2011, 286(24): 21137-21144.
  1. 1. Céleste C, Ionescu M, Robin Poole A, et al. Repeated intraarticular injections of triamcinolone acetonide alter cartilage matrix metabolism measured by biomarkers in synovial fluid. J Orthop Res, 2005, 23(3): 602-610.
  2. 2. Madsen SH, Andreassen KV, Christensen ST, et al. Glucocorticoids exert context-dependent effects on cells of the joint in vitro. Steroids, 2011, 76(13): 1474-1482.
  3. 3. Jaramillo MC, Briehl MM, Crapo JD, et al. Manganese porphyrin, MnTE-2-PyP (5+), acts as a pro-oxidant to potentiate glucocorticoid-induced apoptosis in lymphoma cells. Free Radic Biol Med, 2012. [Epub ahead of print].
  4. 4. Chung YJ, Lee JI, Chong S, et al. Anti-proliferative effect and action mechanism of dexamethasone in human medullary thyroid cancer cell line. Endocr Res, 2011, 36(4): 149-157.
  5. 5. Li WZ, Li WP, Huang DK, et al. Dexamethasone and Aβ25-35 accelerate learning and memory impairments due to elevate amyloid precursor protein expression and neuronal apoptosis in 12-month male rats. Behav Brain Res, 2012, 227(1): 142-149.
  6. 6. Zuloaga DG, Carbone DL, Hiroi R, et al. Dexamethasone induces apoptosis in the developing rat amygdala in an age-, region-, and sex-specific manner. Neuroscience, 2011, 199: 535-547.
  7. 7. Ortega-Ortega Y, García-Latorre EA, Lezama RA, et al. Heat shock protein 60 from Klebsiella pneumoniae protects mononuclear cells from apoptotic cell death induced by dexamethasone. Microb Pathog, 2011, 51(5): 352-359.
  8. 8. Silvestrini G, Ballanti P, Patacchioli FR, et al. Evaluation of apoptosis and the glucocorticoid receptor in the cartilage growth plate and metaphyseal bone cells of rats after high-dose treatment with corticosterone. Bone, 2000, 26(1): 33-42.
  9. 9. Hochberg MC, Altman RD, Brandt KD, et al. Guidelines for the medical management of osteoarthritis. Part II. Osteoarthritis of the knee. American College of Rheumatology Arthritis Rheum, 1995, 38(11): 1541-1546.
  10. 10. Dragoo JL, Danial CM, Braun HJ, et al. The chondrotoxicity of single-dose corticosteroids. Knee Surg Sports Traumatol Arthrosc, 2011, [Epub ahead of print].
  11. 11. Braun HJ, Wilcox-Fogel N, Kim HJ, et al. The effect of local anesthetic and corticosteroid combinations on chondrocyte viability. Knee Surg Sports Traumatol Arthrosc, 2011. [Epub ahead of print].
  12. 12. O’Driscoll SW. The healing and regeneration of articular cartilage. J Bone Joint Surg (Am), 1998, 80(12): 1795-1812.
  13. 13. Hossain MA, Park J, Choi SH, et al. Dexamethasone induces apoptosis in proliferative canine tendon cells and chondrocytes. Vet Comp Orthop Traumatol, 2008, 21(4): 337-342.
  14. 14. Chrysis D, Ritzen EM, Sävendahl L. Growth retardation induced by dexamethasone is associated with increased apoptosis of the growth plate chondrocytes. J Endocrinol, 2003, 176(3): 331-337.
  15. 15. Chrysis D, Zaman F, Chagin AS, et al. Dexamethasone induced apoptosis in proliferative chondrocytes through activation of caspases and suppression of the Akt-phosphatidyinositol 3’-kinase signaling pathway. Endocrinology, 2005, 146(3): 1391-1397.
  16. 16. Jia J, Yao W, Guan M, et al. Glucocorticoid dose determines osteocyte cell fate. FASEB J, 2011, 25(10): 3366-3376.
  17. 17. Lotz M, Hashimoto S, Kühn K. Mechanisms of chondrocyte apoptosis. Osteoarthritis Cartilage, 1999, 7(4): 389-391.
  18. 18. Kim HA, Blanco FJ. Cell death and apoptosis in osteoarthritic cartilage. Curr Drug Targets, 2007, 8(2): 333-345.
  19. 19. Lavrik IN. Regulation of death receptor-induced apoptosis induced via CD95/FAS and other death receptors. Mol Biol (Mosk), 2011, 45(1): 173-179.
  20. 20. 王玉, 孫黎光, 夏春輝. Caspase介導(dǎo)的Fas凋亡途徑. 世界華人消化雜志, 2006, 14(36): 3439-3442.
  21. 21. Naves MA, Pereira RM, Comodo AN, et al. Effect of dexamethasone on human osteoblasts in culture: involvement of β1 integrin and integrin-linked kinase. Cell Biol Int, 2011, 35(11): 1147-1151.
  22. 22. 肖娟, 曹誼林, 范華驊, 等. 體外培養(yǎng)軟骨細(xì)胞凋亡與caspase-3表達(dá)的實(shí)驗(yàn)研究. 中國(guó)修復(fù)重建外科雜志, 2003, 17(5): 418-421.
  23. 23. Wang HQ, Yu XD, Liu ZH, et al. Deregulated miR-155 promotes Fas-mediated apoptosis in human intervertebral disc degeneration by targeting FADD and caspase-3. J Pathol, 2011, 225(2): 232-242.
  24. 24. von Rossum A, Krall R, Escalante NK, et al. Inflammatory cytokines determine the susceptibility of human CD8 T cells to Fas-mediated activation-induced cell death through modulation of FasL and c-FLIP (S) expression. J Biol Chem, 2011, 286(24): 21137-21144.