微管蛋白破坏对软骨细胞代谢功能的影响

发布时间：2018-05-30 00:17

  本文选题：微管蛋白 + 破坏　； 参考：《山西医科大学》2011年硕士论文

【摘要】：目的:探讨微管蛋白破坏对体外关节软骨细胞代谢功能的影响。 方法:2月龄新西兰大白兔8只,处死后取双膝关节全层软骨,采用常规0.4% Pronase酶和0.025%Ⅱ型胶原酶依次消化为软骨细胞培养3天贴壁后,分为对照组和实验组,对照组继续用原代培养基(90%DMEM/F12+10%胎牛血清)培养,实验组在原代培养基中加入微管蛋白破坏剂秋水仙素(终浓度为0.1μmol/l)。加药后第1、2天用Annexin-Ⅴ/PI流式细胞术检测软骨细胞早期凋亡率,加药后第6天细胞爬片HE染色观察细胞形态的改变,加药后第3、6、9天取细胞用实时定量荧光反转录聚合酶链式反应(Real time RT-PCR)法测定软骨细胞Ⅱ型胶原、蛋白多糖以及MMP-13 mRNA的表达量。 结果:实验组第2天软骨细胞早期凋亡率明显高于对照组(P0.05);与对照组比较,实验组第6天软骨细胞呈不规则多角形,胞核深染且分裂相增多,细胞基质减少,实验组第3、6、9天软骨细胞Ⅱ型胶原和蛋白多糖mRNA表达量较对照组均明显降低(P0.05),第6、9天实验组软骨细胞MMP-13mRNA表达较对照组明显增高(P0.01)。 结论:微管蛋白破坏可致软骨细胞早期凋亡,导致体外培养软骨细胞Ⅱ型胶原、蛋白多糖等基质成分合成减少,炎性因子MMP-13的表达增多。
[Abstract]:Objective: to investigate the effect of tubulin damage on the metabolism of articular chondrocytes in vitro. Methods eight New Zealand white rabbits (2 months old) were killed. The chondrocytes were digested into chondrocytes by conventional 0.4% Pronase enzyme and 0.025% collagenase for 3 days. The rabbits were divided into control group and experimental group. The control group was cultured with the primary medium of 90 DMEM / F12% fetal bovine serum, and the experimental group was treated with colchicine (0.1 渭 mol / L). The early apoptosis rate of chondrocytes was detected by Annexin- 鈪，

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