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[1]孫書龍,姜宏,湯曉晨,等.黃芪甲苷對人膝骨關(guān)節(jié)炎退變關(guān)節(jié)軟骨細胞基質(zhì)金屬蛋白酶-1及基質(zhì)金屬蛋白酶-3mRNA表達的影響[J].中醫(yī)正骨,2012,24(10):5-9.
　SUN Shu-long*,JIANG Hong,TANG Xiao-chen,et al.Effect of astragaloside Ⅳ on the expression of matrix metalloproteinase-1 mRNA and matrix metalloproteinase-3 mRNA in the chondrocytes of degenerated joint for patients with knee osteoarthritis[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2012,24(10):5-9.
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黃芪甲苷對人膝骨關(guān)節(jié)炎退變關(guān)節(jié)軟骨細胞基質(zhì)金屬蛋白酶-1 　及基質(zhì)金屬蛋白酶-3mRNA表達的影響()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第24卷
期數(shù):
2012年10期
頁碼:
5-9
欄目:
基礎(chǔ)研究
出版日期:
2012-10-20

文章信息/Info

Title:
Effect of astragaloside Ⅳ on the expression of matrix metalloproteinase-1 mRNA and matrix metalloproteinase-3 mRNA in the chondrocytes of degenerated joint for patients with knee osteoarthritis
作者:
孫書龍1; 姜宏2; 湯曉晨2; 孟祥奇2; 龔正豐2; 王擁軍3; 莫文4
1.南京中醫(yī)藥大學,江蘇 南京 210046;
2.江蘇省蘇州市中醫(yī)醫(yī)院,江蘇 蘇州 215009;
3.上海中醫(yī)藥大學脊柱病研究所,上海 200032;
4.上海中醫(yī)藥大學附屬龍華醫(yī)院,上海 200032
Author(s):
SUN Shu-long*; JIANG Hong; TANG Xiao-chen; MENG Xiang-qi; GONG Zheng-feng; WANG Yong-jun; MO Wen.
*Nanjing University of Chinese Medicine,Nanjing 210046,Jiangsu,China
關(guān)鍵詞:
骨關(guān)節(jié)炎; 膝 軟骨細胞 黃芪甲苷 基質(zhì)金屬蛋白酶1 基質(zhì)金屬蛋白酶3
Keywords:
Osteoarthritis; knee;  Chondrocytes;  AstragalosideⅣ;  Matrix metalloproteinase 1;  Matrix metalloproteinase 3
摘要:
目的:研究黃芪甲苷對人膝骨關(guān)節(jié)炎退變關(guān)節(jié)軟骨細胞基質(zhì)金屬蛋白酶-1及基質(zhì)金屬蛋白酶-3mRNA表達的影響。方法:取膝骨關(guān)節(jié)炎患者膝關(guān)節(jié)軟骨進行細胞培養(yǎng),將培養(yǎng)的軟骨細胞分離傳代后取第3代細胞,通過HE染色、甲苯胺藍染色和Ⅱ型膠原免疫熒光染色進行軟骨細胞鑒定,并進行基質(zhì)金屬蛋白酶-1和基質(zhì)金屬蛋白酶-3免疫熒光染色。觀察軟骨細胞退變情況后將其分為6組,A組不進行干預; B組加入25 mmol·L-1黃芪甲苷; C組加入50 mmol·L-1黃芪甲苷; D組加入75 mmol·L-1黃芪甲苷; E組加入100 mmol·L-1黃芪甲苷; F組加入500 mmol·L-1黃芪甲苷。分別于培養(yǎng)開始后4 h、8 h、24 h、48 h、72 h 5個時間點,采用實時熒光定量聚合酶鏈式反應檢測各組退變軟骨細胞內(nèi)基質(zhì)金屬蛋白酶-1和基質(zhì)金屬蛋白酶-3mRNA表達的情況。結(jié)果:①形態(tài)觀察。原代細胞中梭形細胞和多角形細胞并存,細胞核偏于細胞的一側(cè)而貼近細胞膜,細胞纖絲較長。HE染色細胞質(zhì)為紅色,胞膜呈藍色; 甲苯胺藍染色細胞質(zhì)和胞膜均呈深藍色; Ⅱ型膠原染色陽性為綠色熒光,細胞核采用4',6-二脒基-2-苯基吲哚復染,在不同波段熒光激發(fā)下,胞漿和胞膜可見清晰綠色熒光,細胞核區(qū)域未見明顯綠色熒光,證明培養(yǎng)細胞表達特征性基因Ⅱ型膠原,主要分布在胞漿和細胞膜上,證明所培養(yǎng)的細胞為軟骨細胞。基質(zhì)金屬蛋白酶-1和基質(zhì)金屬蛋白酶-3免疫熒光染色可見清晰綠色熒光,軟骨細胞以多角形多見。②基質(zhì)金屬蛋白酶-1mRNA表達。不同時間點間軟骨細胞基質(zhì)金屬蛋白酶-1mRNA表達水平有差異(F=11.027,P=0.000); 不同組間軟骨細胞基質(zhì)金屬蛋白酶-1mRNA表達水平有差異(F=102.345,P=0.000),A組高于B組、C組、D組、E組(P=0.000; P=0.000; P=0.000; P=0.000),F組高于A組、B組、C組、D組、E組(P=0.009; P=0.000; P=0.000; P=0.000; P=0.000); 時間因素和組間因素存在交互效應(F=8.258,P=0.000)。③基質(zhì)金屬蛋白酶-3mRNA表達。不同時間點間軟骨細胞基質(zhì)金屬蛋白酶-3mRNA表達水平有差異(F=12.316,P=0.000); 不同組間軟骨細胞基質(zhì)金屬蛋白酶-3mRNA表達水平有差異(F=66.112,P=0.000),A組高于B組、C組、D組、E組(P=0.008; P=0.000; P=0.000; P=0.000),F組高于A組、B組、C組、D組、E組(P=0.000; P=0.000; P=0.000; P=0.000; P=0.000); 時間因素和組間因素存在交互效應(F=14.846,P=0.000)。結(jié)論:低濃度的黃芪甲苷能通過抑制退變軟骨細胞合成基質(zhì)金屬蛋白酶-1和基質(zhì)金屬蛋白酶-3,延緩軟骨細胞退變; 濃度過高則會促進退變軟骨細胞合成基質(zhì)金屬蛋白酶-1和基質(zhì)金屬蛋白酶-3,加速軟骨細胞退變。
Abstract:
Objective:To study on the effect of astragaloside Ⅳ on the expression of matrix metalloproteinase-1 mRNA(MMP-1 mRNA)and matrix metalloproteinase-3 mRNA(MMP-3 mRNA)in the chondrocytes of degenerated joint for patients with knee osteoarthritis(KOA).Methods:Knee joint cartilages were fetched out from KOA patients for cell culture,and the third generation cells were chosen from the cultured chondrocytes after isolation and subculture.Chondrocytes identification was made through HE staining,toluidine blue staining and collagen typeⅡimmunofluorescence staining,then immunofluorescent staining was used to detect MMP-1 and MMP-3.The chondrocytes were divided into 6 groups after observing their degeneration conditions.Cells in group A were not intervened,cells in other groups were placed in the culture fluids respectively added with 25 mmol·L-1 astragaloside Ⅳ(group B),50 mmol·L-1 astragalosideⅣ(group C),75 mmol·L-1 astragalosideⅣ(group D),100 mmol·L-1 astragalosideⅣ(group E),and 500 mmol·L-1 astragalosideⅣ(group F).The expression of MMP-1 mRNA and MMP-3 mRNA in the degenerated chondrocytes of the 6 groups were inspected through real-time fluorescence quantitative polymerase chain reaction 4,8,24,48 and 72 hours after the culture respectively.Results:①Morphological observations:the situations as coexistence of spindle cells and polygonal cells,the nucleus located at the side of cells were close to the cell membrane and the longer cell microfibril were shown in primary cells.Red cytoplasm and blue cell membrane were shown after HE staining; cytoplasm and cell membrane were dark blue after toluidine blue staining,collagen typeⅡ-positive cells were shown of green fluorescence after immunofluorescence staining,and nucleus were further processed with counterstaining through 4',6-diamidino-2-phenylindole,then the clear green fluorescence was shown in cytoplasm and cell membrane under fluorescence excitation in different wavebands,while there was not obvious green fluorescence in nuclear regions,which showed that the characteristic gene collagen type Ⅱwere expressed in the culture cells and were mainly distributed in the cytoplasm and cell membrane,then the culture cells were proved to be chondrocytes.Clear green fluorescence was shown after MMP-1 and MMP-3 immunofluorescent staining,and most of chondrocytes were polygonal cells.②MMP-1 mRNA expression:there was statistical difference in MMP-1 mRNA expression levels of chondrocytes among different time points(F=11.027,P=0.000); there was statistical difference in MMP-1 mRNA expression levels of chondrocytes among different groups(F=102.345,P=0.000),and expression level of group A was higher than that of group B,C,D,E respectively(P=0.000; P=0.000; P=0.000; P=0.000; P=0.000)and expression level of group F was higher than that of group A,B,C,D,E respectively(P=0.009; P=0.000; P=0.000; P=0.000; P=0.000); there was interaction between time factor and grouping factor(F=8.258,P=0.000).③MMP-3 mRNA expression:there was statistical difference in MMP-3 mRNA expression levels of chondrocytes among different time points(F=12.316,P=0.000); there was statistical difference in MMP-3 mRNA expression levels of chondrocytes among different groups(F=66.112,P=0.000),and expression level of group A was higher than that of group B,C,D,E respectively(P=0.008; P=0.000; P=0.000; P=0.000)and expression level of group F was higher than that of group A,B,C,D,E respectively(P=0.000; P=0.000; P=0.000; P=0.000; P=0.000); there was interaction between time factor and grouping factor(F=14.846,P=0.000).Conclusion:Low concentration of astragalosideⅣcan delay chondrocytes degeneration through inhibiting the synthesis of MMP-1 and MMP-3 in degenerated chondrocytes,while high concentration of astragalosideⅣcan accelerate chondrocytes degeneration through promoting the synthesis of MMP-1 and MMP-3.

參考文獻/References:

[1] Roach HI,Aigner T,Kouri JB.Chondroptosis:a variant of apoptotic cell death in chondrocytes?[J].Apoptosis,2004,9(3):265-277.
[2] Del Carlo M Jr,Loeser RF.Cell death in osteoarthritis[J].Curr Rheumatol Rep,2008,10(1):37-42.
[3] Malemud CJ,Goldberg VM.Future directions for research and treatment of osteoarthritis[J].Front Biosci,1999,4:D762-D771.
[4] 劉威,趙印震,盧博.分期治療膝關(guān)節(jié)骨性關(guān)節(jié)炎[J].四川中醫(yī),2007,25(8):91-92.
[5] Hardingham T.Extracellular matrix and pathogenic mechanisms in osteoarthritis[J].Curr Rheumatol Rep,2008,10(1):30-36.
[6] Buckwalter JA,Martin JA.Osteoarthritis[J].Adv Drug Deliv Rev,2006,58(2):150-67.
[7] Tetlow LC,Adlam DJ,Woolley DE.Matrix metalloproteinase and proinflammatory cytokine production by chondrocytes of human osteoarthritic cartilage:associations with degenerative changes[J].Arthritis Rheum,2001,44(3):585-594.
[8] 管劍龍,施桂英.基質(zhì)金屬蛋白酶與骨關(guān)節(jié)炎[J].中華風濕病學雜志,2000,4(1):54-55.
[9] Hausser HJ,Decking R,Brenner RE,et al.Testican-1,an inhibitor of pro-MMP-2 activation,is expressed in cartilage[J].Osteoarthritis Cartilage,2004,12(11):870-877.
[10] 李桂葉,張榮富,潘琳,等.骨關(guān)節(jié)炎兔血清中軟骨寡聚基質(zhì)蛋白和基質(zhì)金屬蛋白酶-3水平[J].中華內(nèi)科雜志,2011,50(5):416-419.
[11] 雷載權(quán),陳松育,高學敏.中藥學[M].上海:上海科學技術(shù)出版社,1995:280.

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備注/Memo

備注/Memo:
基金項目:江蘇省中醫(yī)藥管理局計劃項目(HZ07089)
通訊作者:姜宏 E-mail:[email protected]
更新日期/Last Update: 2012-10-20