Zhang Zhengyao, Liu Xiaofeng, Duan Lengxin, Li Xia, Zhang Yang, Zhou Qiuli
Department of Biopharmacy, School of Pharmaceutical Sciences, Jilin University, Jilin, PR China.
Acta Biochim Pol. 2011;58(3):297-302. Epub 2011 Jul 11.
To study the effects of velvet antler polypeptides (VAPs) on osteoarthritic chondrocytes (OCs) in rabbits.
An osteoarthritic rabbit model was established according to Hulth's method. OCs were isolated and cultured for observation of the cell cycle. Cell proliferation was detected by MTT assay and the cell cycle was monitored by flow cytometry. The phenotype was determined by toluidine blue staining as well as immunohistochemical staining for collagen type II. The expression of MMP-1, MMP-3, MMP-13, TIMP-1, and collagen I and X mRNA by chondrocytes was assayed by RT-PCR.
The VAPs had no obvious proliferative effect on OCs and did not affect the cell cycle. However, they significantly reduced the proportion of early apoptotic cells in a dose-dependent manner. Further, VAPs inhibited the expression of collagen I and X mRNA and induced abnormal expression of MMP-1 and MMP-13 mRNA. VAPs had no significant effect on MMP-3 and TIMP-1 mRNA levels. The toluidine blue and collagen type II immunohistochemical staining intensities of VAP-treated chondrocytes were positively correlated with the concentration of VAPs used.
VAPs had no significant effect on OC proliferation and the cell cycle, but did increase the glycosaminoglycan (GAG) and collagen type II expression levels in the extracellular matrix, and down-regulated collagen I and X mRNA expression. Treatment of cartilage cells with VAPs maintained their normal phenotype, inhibited matrix metalloproteinases (MMPs) secretion, kept the balance of cartilage matrix metabolism, and sustained an external environment where the cartilage cells could survive. Moreover, VAPs reduced the proportion of early apoptotic cells, suggesting that they may block the apoptotic pathway in OCs.
研究鹿茸多肽（VAPs）对兔骨关节炎软骨细胞（OCs）的影响。
按照胡尔特方法建立兔骨关节炎模型。分离培养OCs以观察细胞周期。采用MTT法检测细胞增殖，用流式细胞术监测细胞周期。通过甲苯胺蓝染色以及Ⅱ型胶原免疫组化染色确定细胞表型。采用RT-PCR检测软骨细胞中MMP-1、MMP-3、MMP-13、TIMP-1以及Ⅰ型和X型胶原mRNA的表达。
VAPs对OCs无明显增殖作用，不影响细胞周期。然而，它们能以剂量依赖方式显著降低早期凋亡细胞比例。此外，VAPs抑制Ⅰ型和X型胶原mRNA的表达，并诱导MMP-1和MMP-13 mRNA异常表达。VAPs对MMP-3和TIMP-1 mRNA水平无显著影响。VAP处理的软骨细胞的甲苯胺蓝和Ⅱ型胶原免疫组化染色强度与所用VAPs浓度呈正相关。
VAPs对OCs增殖和细胞周期无显著影响，但能增加细胞外基质中糖胺聚糖（GAG）和Ⅱ型胶原表达水平，并下调Ⅰ型和X型胶原mRNA表达。用VAPs处理软骨细胞可维持其正常表型，抑制基质金属蛋白酶（MMPs）分泌，保持软骨基质代谢平衡，维持软骨细胞能够存活的外部环境。此外，VAPs降低早期凋亡细胞比例，表明它们可能阻断OCs中的凋亡途径。
