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目的探讨河蚌葡聚糖(HBP-A)治疗骨关节炎大鼠6周后的含药血清对软骨细胞β-catenin和MMP-13基因表达的影响。方法采用6周龄的雄性SD大鼠,随机分为空白组、模型组、西药组和中药组,每组5只。采用ACLT造模方法建立膝骨关节炎模型,造模后采取相应的干预措施,连续灌胃6周后采含药血清。软骨细胞依次用2.5%、5%、10%浓度的HBP-A含药血清进行体外培养,分为空白组、模型组、西药组和中药组。培养48h后观察细胞的形态变化,采用PCR实验检测各组细胞β-catenin和MMP-13基因的表达。结果形态学观察发现10%含药血清明显促进细胞增殖,同时伴发细胞老化凋亡现象。5%浓度含药血清培养的β-catenin、MMP-13基因表达较2.5%增加,差异有统计学意义(P<0.001)。与空白组比较,2.5%、5%的模型组β-catenin和MMP-13基因表达升高,差异有统计学意义(P<0.001)。与模型组比较,2.5%、5%浓度的西药组和中药组β-catenin、MMP-13基因表达降低,差异有统计学意义(P<0.001)。与西药组比较,2.5%、5%浓度的中药组β-catenin、MMP-13基因表达降低,差异有统计学意义(P<0.001)。结论 HBP-A含药血清通过降低β-catenin和MMP-13基因的表达起到保护软骨细胞的作用。
Objective To investigate the effects of serum containing β-catenin and MMP-13 on the expression of chondrocytes in serum of rats with osteoarthritis treated with HBP-A. Methods Six-week-old male Sprague-Dawley rats were randomly divided into blank group, model group, western medicine group and traditional Chinese medicine group, with 5 rats in each group. Arthroscopic knee osteoarthritis model was established by ACLT modeling method, and corresponding interventions were taken after the model was established. Serum was collected after 6 weeks of continuous gavage. Chondrocytes were cultured in vitro with 2.5%, 5% and 10% HBP-A-containing serum, and divided into blank group, model group, western medicine group and Chinese herbal medicine group. After cultured for 48h, the morphological changes of cells were observed. The expression of β-catenin and MMP-13 in each group were detected by PCR. Results Morphological observation showed that 10% drug-containing serum significantly promoted cell proliferation, accompanied by cell apoptosis. The expression of β-catenin and MMP-13 in 5% drug serum increased more than 2.5%, the difference was statistically significant (P <0.001). Compared with the blank group, the expression of β-catenin and MMP-13 increased in 2.5% and 5% of the model group, with statistical significance (P <0.001). Compared with the model group, the expressions of β-catenin and MMP-13 in the Chinese medicine group and the traditional Chinese medicine group decreased by 2.5% and 5%, respectively, with statistical significance (P <0.001). Compared with the western medicine group, the expressions of β-catenin and MMP-13 in the Chinese herbal medicine group decreased by 2.5% and 5% respectively, with statistical significance (P <0.001). Conclusion HBP-A-containing serum can protect chondrocytes by decreasing the expression of β-catenin and MMP-13.