本文選題：微管蛋白 + 破壞　； 參考：《山西醫(yī)科大學(xué)》2011年碩士論文

【摘要】：目的:探討微管蛋白破壞對體外關(guān)節(jié)軟骨細胞代謝功能的影響。 方法:2月齡新西蘭大白兔8只,處死后取雙膝關(guān)節(jié)全層軟骨,采用常規(guī)0.4% Pronase酶和0.025%Ⅱ型膠原酶依次消化為軟骨細胞培養(yǎng)3天貼壁后,分為對照組和實驗組,對照組繼續(xù)用原代培養(yǎng)基(90%DMEM/F12+10%胎牛血清)培養(yǎng),實驗組在原代培養(yǎng)基中加入微管蛋白破壞劑秋水仙素(終濃度為0.1μmol/l)。加藥后第1、2天用Annexin-Ⅴ/PI流式細胞術(shù)檢測軟骨細胞早期凋亡率,加藥后第6天細胞爬片HE染色觀察細胞形態(tài)的改變,加藥后第3、6、9天取細胞用實時定量熒光反轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)(Real time RT-PCR)法測定軟骨細胞Ⅱ型膠原、蛋白多糖以及MMP-13 mRNA的表達量。 結(jié)果:實驗組第2天軟骨細胞早期凋亡率明顯高于對照組(P0.05);與對照組比較,實驗組第6天軟骨細胞呈不規(guī)則多角形,胞核深染且分裂相增多,細胞基質(zhì)減少,實驗組第3、6、9天軟骨細胞Ⅱ型膠原和蛋白多糖mRNA表達量較對照組均明顯降低(P0.05),第6、9天實驗組軟骨細胞MMP-13mRNA表達較對照組明顯增高(P0.01)。 結(jié)論:微管蛋白破壞可致軟骨細胞早期凋亡,導(dǎo)致體外培養(yǎng)軟骨細胞Ⅱ型膠原、蛋白多糖等基質(zhì)成分合成減少,炎性因子MMP-13的表達增多。
[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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