硫酸软骨素抑制实验性骨关节炎软骨细胞凋亡的体内研究

Inhibition of chondroitin sulfate on apoptosis of chondrocyte in experimental osteoarthritis in vivo

目的探讨硫酸软骨素对骨关节炎软骨细胞凋亡的影响,并研究其可能的作用机制。方法均采用改良Hulth造模法对20只新西兰兔进行造模实验,每组10只,给药组从第4周开始每天2次用5 g/m L硫酸软骨素灌胃,模型组给予等量0.9%Na Cl溶液。给药结束后取兔膝关节软骨组织切片后HE染色,观察组织形态学变化。用Propidium Iodide染色法检测2组兔软骨细胞凋亡情况,Ki67染色法检测细胞增殖。RT-q PCR检测mRNA水平上凋亡因子Caspases-3和凋亡抑制因子Bcl-2的表达变化。结果形态学观察及HE染色结果显示,给药组的病理变化程度较模型组轻。给药组股骨髁、胫骨平台软骨评分显著优于模型组(P<0.01)。给药组凋亡指数明显低于模型组凋亡指数(P<0.01)。给药组细胞的Ki67阳性率明显高于模型组(P<0.001)。给药组中Caspase-3的mRNA水平明显低于模型组(P<0.001)(图3A);而与此相对的,Bcl-2 mRNA水平明显高于模型组(P<0.001)。结论硫酸软骨素能够抑制骨关节炎软骨细胞过度凋亡,并减少凋亡基因Caspase-3的表达和增加凋亡抑制基因Bcl-2的表达。

Objective To explore the inhibitory effect of chondroitin sulfate on apoptosis of chondrocyte in experimental osteoarthritis in vivo and its mechanism. Methods 20 New Zealand white rabbits were established osteoarthritis model by improved Huhh modeling method with 10 rabbits in each group, the treatment group received 5 g/mL chondroitin sulfate by gavage from the 4^th week, twice daily, and the model group received 0. 9% NaCl solution. After treatment, the cartilage tissue of keen was taken out and histomorphology was observed by HE dyeing. The apoptosis of cartilage cells was observed by Propidium Iodide method, and cell proliferation was detected by Ki67 dyeing. The related apoptosis factor of Caspases-3 and Bcl-2 mRNA were detected by RT-qPCR. Results The morphological observation and HE results showed that the pathological change in treatment group was slighter than mondel group. The cartilage scores of condyles of femur and tibial plateau were better than model group （P 〈 0. 01 ）. The apoptosis index in treatment group was lower than model group （ P 〈 0. 01 ）. The Ki67 positive rate in treatment group was higher than model group （ P 〈 0. 001 ）. The levels of Caspase-3 mRNA was lower and Bel-2 mRNA was higher in treatment group than model group（ P 〈 0. 001 ）： Conclusion The chondroitin sulfate could inhibit the excessive osteoarthritis chondrocytes apoptosis , and reduce the expression of apoptosis gene caspase-3, while increase the expression of apoptosis inhibiting gene Bcl-2.