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国家自然科学基金(30470465)

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含14-3-3蛋白抑制肽R18重组腺病毒制备及其在心肌细胞中的表达
2008年
目的:构建并鉴定含14-3-3蛋白抑制肽R18的重组腺病毒,为研究14-3-3蛋白的功能提供基础工具。方法:用同源重组方法构建含14-3-3蛋白抑制肽R18的复制缺陷型腺病毒载体(AdR18),并加以鉴定、扩增,以获得高滴度AdR18病毒液,体外感染乳大鼠心肌细胞,检测目的基因表达。结果:将构建的重组腺病毒载体AdR18感染乳大鼠心肌细胞并表达48h后,蛋白印迹结果显示AdR18感染组有明显R18的表达,对照组无表达。结论:腺病毒载体可高效率导入外源基因在心肌细胞中高表达。
廖文强庞燕于长安李耿温见燕张翼冠李晓辉
关键词:重组腺病毒心肌细胞
三七皂苷对氧化低密度脂蛋白损伤血管内皮细胞保护作用的研究被引量:32
2008年
目的:研究三七皂苷(PNS)对氧化低密度脂蛋白(Ox-LDL)损伤血管内皮细胞的保护作用。方法:体外培养人脐静脉内皮细胞(HUVEC),加入100mg·L-1Ox-LDL造成细胞损伤,同时加入不同浓度PNS进行干预。观察形态学变化;采用MTT法测定细胞活性;比色法检测细胞上清液中乳酸脱氢酶(LDH)的释放量;硝酸还原法和放射免疫法分别测定HUVEC培养上清液中一氧化氮(NO)、内皮素(ET)含量。结果:各剂量PNS可使作用Ox-LDL的HUVEC形态趋于正常。与正常对照组比较,HU-VEC经Ox-LDL作用后,细胞活性显著下降,LDH的释放量显著升高;300、100、30mg·L-1PNS显著升高细胞活性,抑制LDH释放。HUVEC经Ox-LDL作用后,与正常对照组比较,培养上清液中NO含量显著降低,ET含量显著升高;300、100mg·L-1PNS显著升高NO含量,300mg·L-1PNS显著降低ET含量。结论:PNS对Ox-LDL损伤的血管内皮细胞具有一定的保护作用,可调节内皮细胞活性物质NO、ET的分泌。
何翠瑶李晓辉李淑惠何雪峰周胜华
关键词:三七皂苷氧化低密度脂蛋白乳酸脱氢酶
Atherosclerosis lesion is accelerated by persistent systemic inflammation but attenuated by saponins from Panax Notoginseng in rabbits
2008年
Objective: To explore the roles of persistent systemic inflammation in atherosclerosis and the effects of saponins of Panax Notoginseng (PNS) on this process in rabbits. Methods: Thirty rabbits were divided randomly and equally into 6 groups, i.e., control, high-fat diet, inflammation, aspirin, PNS and simple-inflammation group. All the animals except that in control group and simple-inflammation group were fed with high-fat diet for 8 weeks. Based on that, rabbits in inflammation, aspirin and PNS groups were treated with zymosan injection (10 mg/kg, i.p.). Normal saline was given to rabbits in control group. Besides zymosan injection, animals in aspirin and PNS group were administrated with aspirin (12 mg/kg, i.g.) and PNS (120 mg/kg, i.g.) respectively. The animals in simple-inflammation group were treated with zymosan injection (10mg/kg, i.p.) and fed with normal diet. The atherosclerosis lesion in aortas was observed by Sudan IV staining. Serum total cholesterol, triglyceride (TG), TNF-α and activity of post-heparin lipoprotein lipase (LPL) were measured at the end of the 4th and 8th week after an overnight fast. Results: Compared with high-fat diet group, the area of atherosclerosis lesion, serum TG and TNF-α were markedly increased in rabbits of inflammation group, and the activity of LPL was decreased remarkably. Serum TNF-α level was negatively correlated with the activity of post-heparin LPL (r=0.708, P〈0.01). The area of atherosclerosis, serum TG and TNF-α were decreased in aspirin and PNS group compared with that in inflammation group, and the activity of LPL was increased remarkably. Compared with control group, serum TG and TNF-α were markedly increased in simple-inflammation group, while LPL activity was decreased. Atherosclerotic lesion did not occur in simple-inflammation group. Conclusion: Persistent systemic inflammation could accelerate the formation of atherosclerosis lesion in aortas, which partly depend on the decreasing of the activity of post-
Jia Yi Li Xiaohui Liu Ya Zhang Haigang
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