如意珍宝片含药血清对大鼠软骨细胞增殖及分化的影响

目的探讨如意珍宝片含药血清对大鼠软骨细胞增殖及分化的影响。方法以如意珍宝片给大鼠灌胃1周后的含药血清干预软骨细胞，观察不同时间点的细胞形态，用蛋白印迹法检测细胞增殖细胞核抗原（PCNA）蛋白表达情况。进一步采用10ng／ml浓度的白细胞介素（IL）-1β诱导软骨细胞分化，分为空白血清组，IL-1β诱导＋空白血清组，IL-1β诱导＋含药血清组共3组培养，采用实时定量聚合酶链反应（RT—PCR）检测各组细胞的Sox-9和基质金属蛋白酶13（MMP-13）基因的表达，用蛋白印迹法检测各组细胞的B—catenin蛋白表达情况。结果如意珍宝片含药血清能促进软骨细胞的增殖，提高软骨细胞PCNA蛋白及Sox。9基因表达（P〈0．05），但对MMP-13基因和β—catenin蛋白表达无明显影响（P〉0．05）。结论如意珍宝片含药血清具有一定的促进软骨细胞增殖，抑制细胞分化的作用。

Effects of serum containing metabolic ingredients of ruyizhenbao tablets on proliferation and differentiation of chondrocytes in rats

Objective To investigate the effects of serum containing metabolic ingredient from Ruyizhenbao tablets on proliferation and differentiation of chondrocytes in rats. Methods Rats were intragastrically fed with serum containing metabolic ingredients of Ruyizhenbao tablets for 1 week to determine the morphology of chondro- cytes at various time points. Western blotting was employed to examine the expression of proliferating cell nuclear antigen （PCNA）. The 10 ng/ml interleukin-1β （IL-1β） was administered to induce chondrocyte differentiation. Cells were allocated to blank serum group （group I ）, IL-113 plus blank serum group （group lI） and IL-1β plus serum con- taining metabolic ingredients of Ruyizhenbao tablets （group III）, respectively. The mRNA expression of sex determin- ing region Y-box 9 （Sox-9） and matrix metallopeptidase-13 （MMP-13） were detected by real-time polymerase chain re- action. Furthermore, the expression level of β-catenin was examined by western blotting. Results Serum containing metabolic ingredients of Ruyizhenbao tablets promoted chondrocyte proliferation, as evidenced by augmented expres- sion of PCNA and Sox9 mRNA （both P〈0.05）, but not MMP-13 or β-catenin expression （bothP〉0.05）. Conclusion Serum containing metabolic ingredients of Ruyizhenbao tablets promotes chondrocyte proliferation and inhibits chon- drocyte differentiation in rats. Key words] Chondrocytes; Cell proliferation; Differentiation; Ruyizhenbao tablet