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
Objective: To determine the effect of cis-9, trans-1 1-conjugated linoleic acid on the cell cycle of mammary cancer cells (MCF-7) and the possible mechanism of the inhibitory effect of c9,t11-CLA. Methods: Using cell ...Objective: To determine the effect of cis-9, trans-1 1-conjugated linoleic acid on the cell cycle of mammary cancer cells (MCF-7) and the possible mechanism of the inhibitory effect of c9,t11-CLA. Methods: Using cell culture and immunocytochemical techniques, we examined the cell growth, DNA synthesis, expression of PCNA, cyclin A, B1, D1, p16ink4a and p21cip/waf1 of MCF-7 cells at various c9,t11-CLA concentrations (25μM, 50μM, 100μM and 200μM), at 24h and 48h. 96% ethand was used as negative control. Results: The cell growth and DNA synthesis of MCF-7 cells were inhibited by c9,t11-CLA. After treatment with various doses of c9,t11-CLA mentioned above for 8 days, the inhibition frequency was 27.18%, 35.43%, 91.05%, and 92.86%, respectively. Inhibitory effect of c9,t11-CLA on DNA synthesis (except for 25μM, 24h) was demonstrated by significantly less incorporation of 3H-TdR than the negative control (P<0.05 and P<0.01). To further investigate the influence of the cell cycle progression, we found that c9,t11-CLA may arrest the cell cycle of MCF-7 cells. Immunocytochemical staining demonstrated that incubation with different concentration of c9,t11-CLA at various times significantly decreased the expression of PCNA, Cyclin A, B1, D1 in MCF-7 cells compared to the negative control (P<0.01), whereas the expression of p16ink4a and p21cip/waf1, cyclin-dependent kinases inhibitors (CDKI), were increased. Conclusions: The cell growth and proliferation of MCF-7 cells is inhibited by c9,t11-CLA via blocking cell cycle, accompanying reduced expression of cyclin A, B1, D1 and enhanced expression of CDKI (p16ink4a and p21cip/waf1).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This work was supported by the National Natural Science Foundation of China(No.39870661). Phone: (0086-451)-3641309 Fax: (0086-451)-3641253
文摘Objective: To determine the effect of cis-9, trans-1 1-conjugated linoleic acid on the cell cycle of mammary cancer cells (MCF-7) and the possible mechanism of the inhibitory effect of c9,t11-CLA. Methods: Using cell culture and immunocytochemical techniques, we examined the cell growth, DNA synthesis, expression of PCNA, cyclin A, B1, D1, p16ink4a and p21cip/waf1 of MCF-7 cells at various c9,t11-CLA concentrations (25μM, 50μM, 100μM and 200μM), at 24h and 48h. 96% ethand was used as negative control. Results: The cell growth and DNA synthesis of MCF-7 cells were inhibited by c9,t11-CLA. After treatment with various doses of c9,t11-CLA mentioned above for 8 days, the inhibition frequency was 27.18%, 35.43%, 91.05%, and 92.86%, respectively. Inhibitory effect of c9,t11-CLA on DNA synthesis (except for 25μM, 24h) was demonstrated by significantly less incorporation of 3H-TdR than the negative control (P<0.05 and P<0.01). To further investigate the influence of the cell cycle progression, we found that c9,t11-CLA may arrest the cell cycle of MCF-7 cells. Immunocytochemical staining demonstrated that incubation with different concentration of c9,t11-CLA at various times significantly decreased the expression of PCNA, Cyclin A, B1, D1 in MCF-7 cells compared to the negative control (P<0.01), whereas the expression of p16ink4a and p21cip/waf1, cyclin-dependent kinases inhibitors (CDKI), were increased. Conclusions: The cell growth and proliferation of MCF-7 cells is inhibited by c9,t11-CLA via blocking cell cycle, accompanying reduced expression of cyclin A, B1, D1 and enhanced expression of CDKI (p16ink4a and p21cip/waf1).
