健膝丸含药血清对IL-1β作用下兔关节软骨细胞Type-II Collagen、MMP-1、MMP-13蛋白表达的影响

摘要

目的：观察健膝丸含药血清对白细胞介素-1β（IL-1β）作用下体外培养的兔关节软骨细胞表达II型胶原（Coll-II）、基质金属蛋白酶-1（MMP-1）、基质金属蛋白酶-13（MMP-13）的影响，探讨健膝丸防治骨关节炎的作用机制。方法：提取1月龄的兔关节软骨细胞、Ⅰ代细胞用于实验。实验分为空白组、模型组、健膝丸组和抗骨增生组。细胞在培养第4天时，模型组、健膝丸组和抗骨增生组分别加入10 ng·mL-1的IL-1β继续进行培养，分别在培养24h、48h、72h时提取蛋白，并用In-cell western blot法检测其表达情况。结果：各时间点空白组与模型组Coll-II、MMP-1及MMP-13的表达比较，差异均有统计学意义（P <0.05）；健膝丸组较模型组Coll-II的表达在48h、72h时，差异有统计学意义（P <0.05）；MMP-1及MMP-13表达在各时间点比较，差异均有统计学意义（P<0.05）。结论：健膝丸可能在刺激软骨细胞分泌Coll-II，或者减轻对Coll-II的破坏，及减少MMP-1、MMP-13表达等途径发挥作用，从而促进软骨细胞增殖和延缓软骨基质降解，具有保护软骨细胞、延缓软骨基质降解的作用，临床上可作为骨关节炎的治疗用药。%Objective:To observe the inlfuence of serum medicated with Jianxi Pill on the expression of Type-II Collagen,MMP-1 and MMP-13 proteins of IL-1β-affected rabbit articular cartilage cells so as to explore the mechanisms of Jianxi Pill in the treatment of osteoarthritis.Methods:Articular chondrocytes of one-month rabbits were extracted,and the I-generation cells were used in the experiments.The experiment included a blank group,a model group,a Jianxi Pill group and an anti-bone hyperplasia group.At the fourth day of cell culture, 10 ng·mL-1 of IL-1β was added to the model group,the Jianxi Pill group and the anti-bone hyperplasia group. Proteins were extracted from the cultivated groups respectively after 24 hrs,48 hrs and 72 hrs and their expressions were tested with In-cell Western Blot Method.Results:The differences of Coll-II,MMP-1 and MMP-13 between the blank group and the model group at each time were statistically signiifcant (P< 0.05).There was signiifcant difference(P< 0.05) of the expression of Coll-II between the Jianxi Pill group and the model group after 48 hrs and 72 hrs.The differences of the expressions of MMP-1 and MMP-13 at each time were statistically signiifcant (P< 0.05). Conclusion:Jianxi Pill might stimulate chondrocytes to secrete Coll-II or reduce the damage to Coll-II and the expressions of MMP-1 and MMP-13,so as to promote the proliferation of chondrocytes and delay the degradation of cartilage matrix.So Jianxi Pill could protect cartilage cells,retard the degradation of cartilage matrix effect,and be clinically applied on the threatment of osteoarthritis.