發(fā)布時(shí)間：2018-05-06 09:34

  本文選題：低氧 + 骨髓間充質(zhì)細(xì)胞來源軟骨樣細(xì)胞　； 參考：《南昌大學(xué)》2011年碩士論文

【摘要】：研究背景和目的： 軟骨細(xì)胞是構(gòu)成關(guān)節(jié)軟骨的主要基礎(chǔ),但因其高度分化增殖能力低下造成軟骨的再生能力差,使得軟骨損傷后難以自然修復(fù)。干細(xì)胞及其來源軟骨細(xì)胞移植治療解決了軟骨細(xì)胞低增殖力問題,已成為臨床上治療軟骨損傷的有效方法。大量研究發(fā)現(xiàn)體外常氧環(huán)境下培養(yǎng)的細(xì)胞移植進(jìn)入類似關(guān)節(jié)腔內(nèi)缺氧環(huán)境后活性及存活率均下降,Wakitani等研究中認(rèn)為移植細(xì)胞低存活率使得干細(xì)胞分化的軟骨的厚度、機(jī)械強(qiáng)度等均不及正常軟骨,所以認(rèn)為細(xì)胞移植的低存活率是制約這一方法應(yīng)用的因素。綠原酸(chlorogenic acid,CGA)為苯丙素類極性有機(jī)酸,具有強(qiáng)抗氧化能力可抑制氧化應(yīng)急引起的凋亡。綠原酸對缺氧環(huán)境下細(xì)胞內(nèi)活性氧改變引起的凋亡的作用,目前還不清楚。本實(shí)驗(yàn)擬體外誘導(dǎo)骨髓間充質(zhì)細(xì)胞為軟骨樣細(xì)胞,0.1% O2缺氧環(huán)境下模擬關(guān)節(jié)腔內(nèi)的缺氧環(huán)境模型,觀察綠缺氧環(huán)境下原酸對軟骨樣細(xì)胞的作用,探討其對軟骨細(xì)胞的影響及可能機(jī)制。 研究內(nèi)容和方法： 采用體外培養(yǎng)大鼠間充質(zhì)干細(xì)胞,以含0.2mg·L-1 BMP-2的DMEM誘導(dǎo)誘導(dǎo)液培養(yǎng)12天使其成為軟骨樣細(xì)胞后進(jìn)行實(shí)驗(yàn)。分為(A)正常對照組；(B)綠原酸組；(C)0.1% O2缺氧環(huán)境組；(D)綠原酸+0.1% O2缺氧環(huán)境組。進(jìn)而檢測各組Hoechst33258熒光染色檢測細(xì)胞凋亡、細(xì)胞內(nèi)活性氧水平；RT-PCR檢測不同組軟骨細(xì)胞Caspase-3和Bcl-2基因的表達(dá)。 結(jié)果： 與正常對照組比較,0.1%O2缺氧環(huán)境12小時(shí)能明顯造成干細(xì)胞來源軟骨樣細(xì)胞的凋亡(P0.05),綠原酸可降低0.1% O2缺氧引起的軟骨樣細(xì)胞凋亡率(P0.05),活性氧水平下降,Bcl-2表達(dá)增強(qiáng),Caspase-3表達(dá)減弱。 結(jié)論： 綠原酸可抑制0.1%02缺氧引起的內(nèi)皮細(xì)胞凋亡,其作用機(jī)制可能與降低細(xì)胞內(nèi)的活性氧水平,穩(wěn)定細(xì)胞的氧化還原狀態(tài)來保護(hù)線粒體膜電位,促進(jìn)凋亡抑制基因Bcl-2的表達(dá)及抑制Caspase-3的表達(dá)有關(guān)。
[Abstract]:Background and objectives of the study: Chondrocytes are the main base of articular cartilage, but the regeneration ability of cartilage is poor due to its low differentiation and proliferation, which makes it difficult to repair cartilage naturally after injury. Transplantation of stem cells and chondrocytes from chondrocytes has solved the problem of low proliferation of chondrocytes and has become an effective method for the treatment of cartilage injury. A large number of studies have found that the viability and survival rate of cells cultured in normoxic environment after transplantation into the anoxic environment of articular cavity decreased. Wakitani and other studies suggested that the low survival rate of transplanted cells resulted in the thickness of the cartilage of stem cell differentiation. The mechanical strength was not as good as normal cartilage, so the low survival rate of cell transplantation was considered to be a factor restricting the application of this method. Lv Yuan chlorogenic acid CGA is a polar organic acid of phenylpropanoid, which has strong antioxidant ability to inhibit the apoptosis induced by oxidation emergency. The effect of Lv Yuan on apoptosis induced by changes of reactive oxygen species in anoxic environment is unclear. The aim of this experiment was to induce bone marrow mesenchymal cells to be chondroid cells in 0.1% O2 hypoxia environment, and to observe the effect of protonic acid on chondroid cells in green anoxic environment. Objective: to investigate the effect of chondrocytes on chondrocytes and its possible mechanism. Contents and methods of the study: Rat mesenchymal stem cells (MSCs) were cultured in vitro, and 12 angels were cultured into chondroid cells in DMEM induction medium containing 0.2mg L-1 BMP-2. They were divided into two groups: normal control group (B)) Lv Yuan group with 0.1% O2 anoxic environment group and D acid 0.1% O2 anoxic environment group. Apoptosis was detected by Hoechst33258 fluorescence staining and the expression of Caspase-3 and Bcl-2 genes in different chondrocytes were detected by RT-PCR. Results: Compared with the control group, 0.1 O _ 2 hypoxia environment for 12 hours could significantly induce apoptosis of chondroid cells derived from stem cells. Lv Yuan acid could decrease the apoptosis rate of chondroid cells induced by 0.1% O _ 2 hypoxia and increase the expression of Caspase-3 in the chondroid cells induced by 0.1% O _ 2 hypoxia. Conclusion: Lv Yuan can inhibit the apoptosis of endothelial cells induced by anoxia. The mechanism may be related to reducing the level of reactive oxygen species in the cells and stabilizing the redox state of the cells to protect the mitochondrial membrane potential. Promoting the expression of apoptosis suppressor gene Bcl-2 and inhibiting the expression of Caspase-3 are related.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 胡向陽,孟剛,鮑揚(yáng)漪,朱曉梅,汪淵,周青;紫杉醇誘導(dǎo)Hela細(xì)胞凋亡及其與凋亡相關(guān)蛋白的關(guān)系[J];中國藥理學(xué)通報(bào);2004年09期

2 王曉琴;王振華;劉梅;張波;;氧化還原調(diào)控在紫檀芪誘導(dǎo)HeLa細(xì)胞內(nèi)質(zhì)網(wǎng)凋亡途徑中的作用[J];中國藥理學(xué)通報(bào);2010年09期

，

本文編號(hào)：1851800