[目的]探究TGF-β1与GATA3对骨关节炎中软骨细胞钙化的影响。[方法]收集外伤及骨关节炎患者的膝关节软骨组织并分离软骨原代细胞，将软骨细胞分为正常组和研究组。用SB-431542或K-7174干扰软骨细胞中TGF-β1/GATA3的表达。用钙化诱导培养基使软骨细胞发生钙化。通过蛋白免疫印迹实验和实时荧光定量PCR实验分析软骨细胞中TGF-β1/GATA3的表达。通过茜素红染色统计细胞钙化水平。[结果]与正常组软骨细胞比较，OA组患者软骨细胞TGF-β1和GATA3的表达水平和钙化水平显著增加(1。01±0。02 vs 2。98±0。03;0。91±0。05 vs 2。33± 0。11;P＜0。05);与 PBS 组比较，K-7174 组软骨细胞的ALP 和 COLX 的表达水平(2。18±0。05 vs 0。91±0。06;2。83± 0。03 vs 0。46±0。01)以及钙化细胞数量显著降低(0。63±0。03 vs 0。25±0。01;P＜0。05);与 PBS 组比较，SB-431542组软骨细胞的ALP和COLX的表达水平(1。01±0。02 vs 0。46±0。05;1。05±0。02 vs 0。43±0。02)以及钙化细胞数量显著降低(P＜0。05);抑制TGF-β1后，GATA3表达水平显著降低(0。99±0。06 vs 0。51±0。02;P＜0。05)。[结论]在骨关节炎患者的软骨组织中，TGF-β1和GATA3表达水平增加，TGF-β1能够提高骨关节炎患者的软骨细胞钙化水平，并且这一过程与调控GATA3表达密切相关。
TGF-β1 regulates GAT A3 to promote chondrocyte calcification in osteoarthritis
[Objective]To investigate the effect of transforming growth factor-β1(TGF-β1)and GATA3 on chondrocyte cal-cification in osteoarthritis.[Method]Primary chondrocytes were isolated from knee cartilage tissues of patients with trauma and osteoarthritis.The chondrocytes were divided into normal group and study group.SB-431542 or K-7174 was used to interfere with TGF-β1/GATA3 expression in chondrocytes.Calcification occurs in chondrocytes using calcification-inducing medium.The expression of TGF-β1/GATA3 in chondrocytes was analyzed by Western Blot and real-time fluorescence quantitative PCR.The level of cellular calcification was counted by alizarin red staining.[Result]Compared with normal chondrocytes,the expression levels of TGF-β1 and GATA3 and the calcification level of chondrocytes in OA patients were significantly increased(1.01±0.02 vs 2.98±0.03;0.91±0.05 vs 2.33±0.11;P＜0.05).Compared with PBS group,the expression levels of ALP and COLX in K-7174 group(2.18±0.05 vs 0.91±0.06;2.83±0.03 vs 0.46±0.01)and the number of calcified cells(0.63±0.03 vs 0.25±0.01;P＜0.05).Compared with the PBS group,the expression levels of ALP and COLX in the SB-431542 group(1.01±0.02 vs 0.46±0.05;1.05±0.02 vs 0.43±0.02)and the number of calcified cells decreased significantly(P＜0.05).After TGF-β1 inhibition,the expression of GATA3 was significantly decreased(0.99±0.06 vs 0.51 ±0.02;P＜0.05).[Conclusion]The expression levels of TGF-β1 and GATA3 are increased in the cartilage of patients with osteoarthritis.TGF-β1 can increase the calcification level of chondrocytes in patients with osteoarthritis,and this process is closely related to the regulation of GATA3 expression.
TGF-β1osteoarthritischondrocytesGATA3calcificationbone and joint lesionstrabecular bonedegenera-tion of cartilage
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