共轭亚油酸是具有一定功能的多不饱和脂肪酸,但反式脂肪酸的功能至今仍有争议,本研究拟探讨实验室自制的t9,t11-CLA对氧化损伤的人脐静脉内皮细胞(HUVECs)的保护作用并对其可能机理进行解析。采用MTT法分析t9,t11-CLA对氧化损伤HUVECs...共轭亚油酸是具有一定功能的多不饱和脂肪酸,但反式脂肪酸的功能至今仍有争议,本研究拟探讨实验室自制的t9,t11-CLA对氧化损伤的人脐静脉内皮细胞(HUVECs)的保护作用并对其可能机理进行解析。采用MTT法分析t9,t11-CLA对氧化损伤HUVECs的保护作用,通过流式细胞术检测t9,t11-CLA对细胞凋亡及线粒体膜电位变化的影响,采用实时荧光定量PCR和caspase蛋白试剂盒检测与细胞凋亡相关的caspase蛋白及基因变化情况。结果表明,t9,t11-CLA能显著增强HUVECs抵御氧化损伤的能力,缓解氧化损伤造成的细胞膜电位下降、细胞凋亡比例上升、caspase-8、-3活性激活及caspase-8、-3 m RNA表达量的上调,说明t9,t11-CLA可能通过与线粒体途径相关的caspase依赖的抗凋亡机制保护了血管内皮细胞免受氧化损伤。展开更多
Considering the results of our previous research that conjugated linoleic acid mixture-paclitaxel （CLA-mixture-PTX） possesses anti-tumor activity against melanoma and brain glioma, the purpose of this study was to i...Considering the results of our previous research that conjugated linoleic acid mixture-paclitaxel （CLA-mixture-PTX） possesses anti-tumor activity against melanoma and brain glioma, the purpose of this study was to investigate the potential anti-tumor efficacy of cis-9, trans- 1 1-conjugated linoleic acid-paclitaxel （c9, tl 1-CLA-PTX） and trans- 1 O, cis- 12-conjugated linoleic acid-paclitaxel （tl0, c12-CLA-PTX） on MCF-7 breast cancer cell line in vitro and in vivo. The in vitro cytotoxicity, apoptosis induction effect and cell cycle arresting effect of c9, t1 1-CLA-PTX and t10, c12-CLA-PTX were investigated. The in vitro cellular uptake of c9, tl 1-CLA-PTX and tl0, cl2-CLA-PTX in MCF-7 cells were also analyzed. Besides, the anti-tumor activity of c9, tl 1-CLA-PTX and tl0, cl2-CLA-PTX was evaluated in MCF-7 tumor bearing nude mice in vivo. The in vitro cytotoxicity results showed that the value of ICs0 of the tl 0, c l2-CLA-PTX is （0.17±0.02） μM, compared with that of （1.08±0.15） μM in CLA-mixture-PTX and （6.50±1.20） μM in c9, tl 1-CLA-PTX treatment group （P〈0.01）. Both tl0, cl2-CLA-PTX and c9, t l 1-CLA-PTX increased the percentage of total apoptotic cells compared with that of control （P〈0.01）. And the rank of apoptosis induction efficacy was t 10, c 12-CLA-PTX〉CLA-mixture-PTX〉c9, t 11-CLA-PTX （P〈0.01）. Compared with untreated cells, the tl0, c12-CLA-PTX and c9, tl 1-CLA-PTX arrested cell cycle progression at the S and G2-M phase. The amount of cellular uptake of t 10, c 12-CLA-PTX was significantly higher than that of CLA-mixture-PTX （P〈0.01）, which was significantly higher than that of c9, t1 1-CLA-PTX （P〈0.01）. The rank of in vivo anti-tumor activity was tl0, c12-CLA-PTX〉CLA-mixture-PTX〉 c9, t1 1-CLA-PTX （P〈0.01）. In conclusion, our study demonstrated that both tl0, cl2-CLA-PTX and c9, tl 1-CLA-PTX has significant anti-tumor activity in MCF-7 cell line. And while c9, tl 1-CLA-PTX showed weaker inhibitory effect than CLA-mixture-PTX, stronger inhibitory effect was presented by t10, c12-CLA-PTX, which could be a promising alternative for CLA-mixture-PTX.展开更多
文摘共轭亚油酸是具有一定功能的多不饱和脂肪酸,但反式脂肪酸的功能至今仍有争议,本研究拟探讨实验室自制的t9,t11-CLA对氧化损伤的人脐静脉内皮细胞(HUVECs)的保护作用并对其可能机理进行解析。采用MTT法分析t9,t11-CLA对氧化损伤HUVECs的保护作用,通过流式细胞术检测t9,t11-CLA对细胞凋亡及线粒体膜电位变化的影响,采用实时荧光定量PCR和caspase蛋白试剂盒检测与细胞凋亡相关的caspase蛋白及基因变化情况。结果表明,t9,t11-CLA能显著增强HUVECs抵御氧化损伤的能力,缓解氧化损伤造成的细胞膜电位下降、细胞凋亡比例上升、caspase-8、-3活性激活及caspase-8、-3 m RNA表达量的上调,说明t9,t11-CLA可能通过与线粒体途径相关的caspase依赖的抗凋亡机制保护了血管内皮细胞免受氧化损伤。
基金National Natural Science Foundation of China (Grant No.81172992)the National Basic Research Program of China (973 Program,Grant No.2013CB932501)Innovation Team of Ministry of Education (Grant No.BMU20110263)
文摘Considering the results of our previous research that conjugated linoleic acid mixture-paclitaxel （CLA-mixture-PTX） possesses anti-tumor activity against melanoma and brain glioma, the purpose of this study was to investigate the potential anti-tumor efficacy of cis-9, trans- 1 1-conjugated linoleic acid-paclitaxel （c9, tl 1-CLA-PTX） and trans- 1 O, cis- 12-conjugated linoleic acid-paclitaxel （tl0, c12-CLA-PTX） on MCF-7 breast cancer cell line in vitro and in vivo. The in vitro cytotoxicity, apoptosis induction effect and cell cycle arresting effect of c9, t1 1-CLA-PTX and t10, c12-CLA-PTX were investigated. The in vitro cellular uptake of c9, tl 1-CLA-PTX and tl0, cl2-CLA-PTX in MCF-7 cells were also analyzed. Besides, the anti-tumor activity of c9, tl 1-CLA-PTX and tl0, cl2-CLA-PTX was evaluated in MCF-7 tumor bearing nude mice in vivo. The in vitro cytotoxicity results showed that the value of ICs0 of the tl 0, c l2-CLA-PTX is （0.17±0.02） μM, compared with that of （1.08±0.15） μM in CLA-mixture-PTX and （6.50±1.20） μM in c9, tl 1-CLA-PTX treatment group （P〈0.01）. Both tl0, cl2-CLA-PTX and c9, t l 1-CLA-PTX increased the percentage of total apoptotic cells compared with that of control （P〈0.01）. And the rank of apoptosis induction efficacy was t 10, c 12-CLA-PTX〉CLA-mixture-PTX〉c9, t 11-CLA-PTX （P〈0.01）. Compared with untreated cells, the tl0, c12-CLA-PTX and c9, tl 1-CLA-PTX arrested cell cycle progression at the S and G2-M phase. The amount of cellular uptake of t 10, c 12-CLA-PTX was significantly higher than that of CLA-mixture-PTX （P〈0.01）, which was significantly higher than that of c9, t1 1-CLA-PTX （P〈0.01）. The rank of in vivo anti-tumor activity was tl0, c12-CLA-PTX〉CLA-mixture-PTX〉 c9, t1 1-CLA-PTX （P〈0.01）. In conclusion, our study demonstrated that both tl0, cl2-CLA-PTX and c9, tl 1-CLA-PTX has significant anti-tumor activity in MCF-7 cell line. And while c9, tl 1-CLA-PTX showed weaker inhibitory effect than CLA-mixture-PTX, stronger inhibitory effect was presented by t10, c12-CLA-PTX, which could be a promising alternative for CLA-mixture-PTX.
