摘要
【目的】探讨姜黄素通过Wnt/β-catenin信号通路对软骨细胞增殖的影响。【方法】取4周龄雌雄新西兰大白兔各1只,采用酶消化法构建体外软骨细胞培养体系;应用形态学观察、甲苯胺蓝染色方法进行细胞形态学鉴定;采用细胞计数试剂盒法(CCK-8法)检测不同浓度姜黄素给药24 h后对软骨细胞增殖的影响;将终浓度为5μmol/L、10μmol/L的姜黄素分别作用于软骨细胞12 h、24 h后,采用Western blot法检测Wnt/β-catenin信号通路中Wnt 2、GSK-3β、β-catenin蛋白表达情况。【结果】当姜黄素的浓度为10μmol/L时,姜黄素对软骨细胞的促增殖作用最为明显。姜黄素能够促进Wnt/β-catenin信号通路中Wnt 2及β-catenin蛋白表达量,且10μmol/L组较5μmol/L组、24 h组较12 h组的表达量更高,24 h两浓度组与对照组比较,差异均有统计学意义(P<0.05),但12 h的两浓度组与对照组比较,差异无统计学意义(P>0.05),而GSK-3β结果则正好与上述2种蛋白表达结果相反。【结论】姜黄素对软骨细胞具有一定的保护作用,是中药姜黄发挥抗骨关节炎作用的有效成分之一。
Objective To investigate the effect of curcumin on the proliferation of chondrocytes through Wnt/β-catenin signaling pathway. Methods One male and one female New Zealand rabbit with 4 weeks old were used for the experiment. The enzyme digestion method was adopted to construct cartilage cell culture system in vitro.Morphological observation and toluidine blue staining method were used for the identification of cell morphology.CCK-8 assay was used to observe the influence of various concentrations of curcumin on the proliferation of chondrocytes after treatment for 24 h. After cartilage cells were cultured with curcumin at final concentration of5, 10 μmol/L respectively for 12, 24 h, Western blot method was used to detect the expression of Wnt 2,GSK-3,β-catenin proteins associated with Wnt/βcatenin signaling pathway. Results When the concentration of curcumin was 10 μmol/L,the effect of curcumin on the proliferation of chondrocytes was the strongest. Curcumin promoted the expression of Wnt 2 and β-catenin proteins associated with Wnt/β-catenin signaling pathway：10μmol/L curcumin group had higher expression levels than 5 μmol/L curcumin（P 0.05）, and curcumin medication for 24 h resulted into higher expression than medication for 12 h,the differences being statistically significant（P 0.05）. The expression levels of Wnt 2 and β-catenin protein in chondrocytes treated with curcumin5,10 μmol/L for 24 h differed from those in the control group（P 0.05）,while the levels in chondrocytes treated with curcumin 5, 10 μmol/L for 12 h did not differ from those in the control group（P 0.05）. The resulst for GSK-3β were in contrast to those of Wnt 2 and β-catenin proteins. Conclusion Curcumin has protective effect on chondrocytes,and is one of the active anti-osteoarthritis ingredients of Rhizoma Curcumae Longae.
出处
《广州中医药大学学报》
CAS
2017年第1期90-95,共6页
Journal of Guangzhou University of Traditional Chinese Medicine
基金
江苏省自然科学基金青年基金资助项目(编号:BK20151007)
南京市医学科技发展项目(编号:YKK13214)
南京中医药大学骨伤修复与重建新技术实验室专项资金资助项目
