论文部分内容阅读
目的以大鼠骨髓间充质干细胞(MSCs)为研究对象,在体外培养过程中观察氧化低密度脂蛋白(ox-LDL)对MSCs增殖、干细胞表面标志Oct-4表达的影响。方法将体外培养的MSCs分为4组:对照组,不加任何干预因素;Ox-LDL组,在培养基中分别加入不同浓度(1、5、10、20μg/mL)的ox-LDL;Ox-LDL+NAC组,采用抗氧化剂乙酰半胱氨酸(NAC)预处理后再加入ox-LDL;阴性对照组,在培养基中加入对应浓度的天然低密度脂蛋白(nLDL)。在不同培养时间点收集细胞计数并绘制生长曲线;蛋白免疫印迹法(Western blot)检测干细胞特异表面标记Oct-4表达水平;电子顺磁共振技术检测氧化活性产物(ROS)产量。结果 ox-LDL抑制MSCs的增殖,浓度越大细胞增殖抑制越明显,当浓度大于5μg/mL时细胞出现凋亡,抗凋亡蛋白Bcl-2表达水平减弱;ox-LDL干预后Oct-4的表达水平显著减弱,且与干预浓度呈正相关;ox-LDL干预后ROS的产量明显升高,而nLDL对ROS生成没有影响。结论 ox-LDL抑制MSCs的体外增殖和Oct-4表达,可能与ROS增加有关。
OBJECTIVE: To investigate the effects of ox-LDL on the proliferation of MSCs and the expression of Oct-4 on the surface of stem cells in rat bone marrow mesenchymal stem cells (MSCs). Methods MSCs cultured in vitro were divided into 4 groups: control group, without any intervention; Ox-LDL group, ox-LDL with different concentration (1,5,10,20μg / mL) -LDL + NAC group. Ox-LDL was pretreated with antioxidant NAC. Negative control group, the corresponding concentration of native LDL (nLDL) was added into the culture medium. Cell counts were collected at different culture time points and growth curves were drawn. Western blot was used to detect the expression of Oct-4 on specific surface markers of stem cells. Electron paramagnetic resonance (ROS) was used to detect the production of ROS. Results Ox-LDL inhibited the proliferation of MSCs. The greater the concentration of ox-LDL, the more obvious the inhibition of cell proliferation was. When the concentration of ox-LDL was more than 5μg / mL, apoptosis occurred and the expression of anti-apoptotic protein Bcl- The expression level of ROS was significantly weakened, and positively correlated with the concentration of intervention. The production of ROS increased obviously after ox-LDL intervention, while nLDL had no effect on the production of ROS. Conclusion Ox-LDL may inhibit the proliferation and the expression of Oct-4 of MSCs in vitro, which may be related to the increase of ROS.