中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
9期
1550-1552
,共3页
郝亚荣%陈宣银%邱波%周建林
郝亞榮%陳宣銀%邱波%週建林
학아영%진선은%구파%주건림
血管内皮生长因子%骨关节炎%软骨细胞%诱导型一氧化氮合成酶
血管內皮生長因子%骨關節炎%軟骨細胞%誘導型一氧化氮閤成酶
혈관내피생장인자%골관절염%연골세포%유도형일양화담합성매
Vascular endothelial growth factor%Osteoarthritis%chondrocyte%Inducible nitric oxide synthase
目的 观察血管内皮生长因子(VEGF)对体外培养的关节软骨细胞诱导型一氧化氮合酶(iNOS)表达的影响。方法 体外培养SD乳鼠关节软骨细胞,用白细胞介素(IL)-1β诱导的方法建立骨关节炎(OA)体外模型,实验分为4组,每组加入不同处理因素进行干预,A组:(正常对照组)不加任何处理因素;B组:10 μg/L VEGF;C组:10 μg/L IL-1β;D组:10 μg/L VEGF+ 10 μg/LIL-1β。采用实时荧光定量PCR( Real Time PCR)检测iNOS mRNA的表达,采用蛋白免疫印迹法( Western blot)检测iNOS蛋白的表达。结果 iNOS mRNA的表达:A组iNOS mRNA无表达,B组(9.64±1.64)、C组(17.27±2.01)及D组(28.93±6.63),3组的iNOS mRNA表达量显著升高,进一步组间比较,D组软骨细胞iNOS的mRNA表达水平明显高于B组(P＜0.01)及C组(P＜0.05),C组软骨细胞iNOS的mRNA表达水平高于B组(P＜0.05)。iNOS蛋白的表达:A组iNOS蛋白无表达,B组(0.44±0.12)、C组(0.74±0.07)及D组(1.38±0.38),3组的iNOS蛋白表达量显著升高,进一步组间比较,D组软骨细胞iNOS的蛋白表达水平明显高于B组(P＜0.01)及C组(P＜0.05),C组软骨细胞iNOS的mRNA表达水平高于B组(P＜0.01)。结论 在OA的发病过程中,VEGF可能通过上调软骨细胞iNOS的表达发挥重要作用。
目的 觀察血管內皮生長因子(VEGF)對體外培養的關節軟骨細胞誘導型一氧化氮閤酶(iNOS)錶達的影響。方法 體外培養SD乳鼠關節軟骨細胞,用白細胞介素(IL)-1β誘導的方法建立骨關節炎(OA)體外模型,實驗分為4組,每組加入不同處理因素進行榦預,A組:(正常對照組)不加任何處理因素;B組:10 μg/L VEGF;C組:10 μg/L IL-1β;D組:10 μg/L VEGF+ 10 μg/LIL-1β。採用實時熒光定量PCR( Real Time PCR)檢測iNOS mRNA的錶達,採用蛋白免疫印跡法( Western blot)檢測iNOS蛋白的錶達。結果 iNOS mRNA的錶達:A組iNOS mRNA無錶達,B組(9.64±1.64)、C組(17.27±2.01)及D組(28.93±6.63),3組的iNOS mRNA錶達量顯著升高,進一步組間比較,D組軟骨細胞iNOS的mRNA錶達水平明顯高于B組(P＜0.01)及C組(P＜0.05),C組軟骨細胞iNOS的mRNA錶達水平高于B組(P＜0.05)。iNOS蛋白的錶達:A組iNOS蛋白無錶達,B組(0.44±0.12)、C組(0.74±0.07)及D組(1.38±0.38),3組的iNOS蛋白錶達量顯著升高,進一步組間比較,D組軟骨細胞iNOS的蛋白錶達水平明顯高于B組(P＜0.01)及C組(P＜0.05),C組軟骨細胞iNOS的mRNA錶達水平高于B組(P＜0.01)。結論 在OA的髮病過程中,VEGF可能通過上調軟骨細胞iNOS的錶達髮揮重要作用。
목적 관찰혈관내피생장인자(VEGF)대체외배양적관절연골세포유도형일양화담합매(iNOS)표체적영향。방법 체외배양SD유서관절연골세포,용백세포개소(IL)-1β유도적방법건립골관절염(OA)체외모형,실험분위4조,매조가입불동처리인소진행간예,A조:(정상대조조)불가임하처리인소;B조:10 μg/L VEGF;C조:10 μg/L IL-1β;D조:10 μg/L VEGF+ 10 μg/LIL-1β。채용실시형광정량PCR( Real Time PCR)검측iNOS mRNA적표체,채용단백면역인적법( Western blot)검측iNOS단백적표체。결과 iNOS mRNA적표체:A조iNOS mRNA무표체,B조(9.64±1.64)、C조(17.27±2.01)급D조(28.93±6.63),3조적iNOS mRNA표체량현저승고,진일보조간비교,D조연골세포iNOS적mRNA표체수평명현고우B조(P＜0.01)급C조(P＜0.05),C조연골세포iNOS적mRNA표체수평고우B조(P＜0.05)。iNOS단백적표체:A조iNOS단백무표체,B조(0.44±0.12)、C조(0.74±0.07)급D조(1.38±0.38),3조적iNOS단백표체량현저승고,진일보조간비교,D조연골세포iNOS적단백표체수평명현고우B조(P＜0.01)급C조(P＜0.05),C조연골세포iNOS적mRNA표체수평고우B조(P＜0.01)。결론 재OA적발병과정중,VEGF가능통과상조연골세포iNOS적표체발휘중요작용。
Objective To investigate the influence of vascular endothelial growth factor (VEGF) on inducible nitric oxide synthase (iNOS) expression in rat articular chondrocytes in vitro. Methods Chondrocytes were isolated and cultured. The model of osteoarthritis (OA) was induced by IL-1β in vitro.Then the experiment was divided into 4 groups: group A ( control ) : without any disposal; group B :10 μg/L VEGF; group C: 10 μg/L 1L-1β; group D: 10 μg/L VEGF+1O μg/L IL-1β. The mRNA and protein expression of iNOS was detected by using real-time polymerase chain reaction, and Western blotting respectively. Results The iNOS mRNA expression levels in group B (9. 64 ± 1.64), group C ( 17. 27 ±2. 01 ) and group D (28. 93 ± 6. 63) were higher than in group A ( control group) significantly, and there was significant difference between groups B and C, B and D, C and D ( B vs C, P ＜ 0. 05 ; B vs D, P ＜0. 01 ; C vs D,P ＜0. 05). As compared with group A, the expression of iNOS protein in group B (0. 44 ±0. 12), group C (0. 74 ±0.07) and group D (1.38 ±0. 38) was significantly increased. There was significant difference between groups B and C, B and D, C and D (B vs C,P＜0.01; B vs D,P＜0.01; C vs D,P ＜ 0. 05 ). Conclusion VEGF significantly improved the expression of iNOS in rat articular chondrocytes. VEGF plays an important role during the development of OA.
