摘要
目的研究黄芪注射液对正常心肌细胞胞外信号调节激酶（ERK）信号转导的影响。方法心肌细胞取材SpragueDawley大鼠。采用培养的原代正常心肌细胞进行研究。量效关系研究分为5组。细胞培养液DMEM中加入黄芪注射液，其终浓度分别为0、20、100、200、400ml／L培养液。37~C培养2h后取细胞进行ERK和一氧化氮合酶（NOS）蛋白表达检测。时效关系研究分为两组。A组为黄芪组，细胞培养液DMEM中加入黄芪注射液。B组为抑制剂组，细胞培养液DMEM中加入黄芪注射液和ERK抑制剂PD98059。37℃孵育，分别于0、5、15、30、60、120min时取细胞进行Western印迹检测ERK和NOS蛋白表达。结果量效关系研究显示黄芪注射液终浓度分别为0、20、100、200、400ml／L培养液时心肌细胞ERK表达没有明显的变化。但NOS表达随黄芪浓度增加有减弱的趋势。时效研究显示心肌细胞加入黄芪注射液培养后，ERK和NOS的表达随时间呈增加趋势。ERK抑制剂PD98059可减弱ERK和NOS的表达。结论黄芪注射液对心肌细胞ERK和NOS信号表达有调节作用，随时间增加呈增强趋势，但随浓度升高呈减弱趋势。
Objective To investigate the dose-effect relationship and time-effect relationship of astragalus injection on the protein expression of nitric-oxide synthase （NOS） and extraeellular signal- regulated kinase （ ERK ） in euhured cardiac myocytes. Methods （ 1 ） Ventrieular myocytes of Sprague- Dawley rats were isolated, euhured, and incubated with astragalus injection of the concentrations of 0, 20, 100, 200, or 400 ml/L for 8 days. Cell growth curve was drawn. Automatic biochemical analyzer was used to detect the level of ereatine kinase （CK）. （2） Western blotting was used 2 h after incubation to observe the protein expression of ERK and nitric oxide synthase （NOS）. （3） Myocytes were incubated with astragalus injection （200 ml/L） or with astragalus and PD98059, an ERK inhibitor, for 5,15,30,60, and 120 min respectively, and Western blotting was used to detect the protein expression of ERK and NOS. Results Astragalus of different concentrations showed no effect on the cell proliferation. Only the CK level of the astragalus 200 ml/L group was significantly higher than that of the control group （ P 〈 0. 05 ）. Astragalus showed no dose-dependent effect on the ERK expression, and dose-dependently reduced the NOS expression level. 15 min after the co-incubation with astragalus 200 ml/L the ERK level of the myocytes decreased transiently, and then began to increase gradually since 15 min after the co-incubation; the NOS level began to increase since 60 min after the co-incubation （ P 〈 0. 05 ）. After the addition of PD98059 the ERK expression significantly decreased （ P 〈 0. 05 ）. Conclusion Astragalus injection has modulation effect on ERK and NOS expression. This effect is enhanced time-dependently at appropriate Astragalus concentration, but attenuated by high dose Astragalus injection.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2008年第19期1359-1362,共4页
National Medical Journal of China
基金
国家自然科学基金资助项目（30572427）
关键词
黄芪
信号传递
一氧化氮合酶
蛋白激酶类
Astragalus membranaceus
Signal transduction
Nitric-oxide synthase
Proteinkinases
