| 摘要: |
| 摘要：目的：通过体外细胞模型实验，研究抑制Pannexin3是否能够治疗骨性关节炎。方法：收集因骨性关节炎行全膝关节置换术患者的关节软骨组织，建立细胞模型。实验组用siRNA+脂质体转染细胞，对照组用NC+脂质体转染细胞。通过RT-PCR及Western-blot检测细胞内MMP-1及MMP-13的mRNA水平及其蛋白表达情况。最后数据用独立样本T检验处理。结果：软骨细胞培养约14天（第一代）及8天（第二代）可长满瓶底80%-90%，前2代较少出现分化，可被II型胶原蛋白免疫化学法将其染成棕黄色，在转染24小时后约80%细胞内可见荧光表达。实验组细胞中MMP-1及MMP-13的mRNA含量及蛋白水平明显低于对照组，P<0.05，差异有统计学意义。结论：抑制Pannexin3基因可抑制骨性关节炎软骨细胞中MMP-1、MMP-13mRNA及其蛋白表达。Pannexin3可能是骨性关节炎潜在治疗靶点。 |
| 关键词: 骨性关节炎 软骨细胞 基质金属蛋白酶1 基质金属蛋白酶13 泛连接蛋白3 |
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| Study the effect of targeted therapy of Pannexin3 on osteoarthritis with cell culture experimental |
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| Abstract: |
| [] 【】 BO Objective: 窗体顶端
[Abstract] Objective:The present study was designed to determine whether inhibition of Pannexin3 is effective in the treatment of osteoarthritis in an in vitro cell model established.
Methods:Articular cartilage tissue of patients undergoing total knee arthroplasty for osteoarthritis was collected and a cell model was established. Cells in the experimental group were transfected with siRNA and those in the control group were transfected with NC by?lipofectamine. Expression levels of MMP-1 and MMP-13 mRNAs and proteins in cells were detected by RT-PCR and Western-blot, respectively. Data obtained were analyzed?with?independent-samples?T test.
Results:Following about 14 days (first generation) and 8 days (second generation) incubation, chondrocytes could adhere to 80%-90% of the bottom of the bottle. The first two generations were less differentiated and could be stained yellow brown by type II collagen using immunohistochemistry. After 24 hours of transfection, fluorescence expression was observed in about 80% of the observed cells.The mRNA and protein levels of MMP-1 and MMP-13 in the experimental group were significantly decreased compared with that of the control group and the differences were statistically significant (p<0.05).
Conclusion:Inhibition of Pannexin3 gene resulted in the decreased expression of MMP-1, MMP-13 mRNAs and proteins in osteoarthritis chondrocytes. Pannexin3 may serve as a potential therapeutic target for the treatment of osteoarthritis. |
| Key words: Osteoarthritis Chondrocytes Matrix Metalloproteinase 1 Matrix Metalloproteinase - 13 Pannexin3 |