尾加压素对人单核巨噬细胞DNA、RNA及蛋白质合成的影响  

Effects of urotensin on synthesis of DNA, RNA and protein of human monocyte macrophages

作　　者：钱云峰[1] 石向东[1] 李志梁[1] 

机构地区：[1]南方医科大学珠江医院心内科,广东省广州市510282

出　　处：《中国临床康复》2005年第39期101-103,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家重点基础发展项目资助(973子课题J200056905)~~

摘　　要：目的:观察新近发现最强的缩血管活性肽之一的尾加压素对体外培养的人单核巨噬细胞DNA,RNA及蛋白质的合成过程的影响。方法:实验于2004-10/12在南方医科大学心内科实验室及广州军区总医院中心实验室完成。取健康自愿者50mL新鲜静脉血,用聚乙酰胺吡咯烷酮包被的硅胶颗粒液(d=1.130)分离单核细胞。采用24孔塑料培养板,每孔加入有巨噬细胞特性、浓度为2×105个/mL细胞0.5mL,随机分成4组。分别加入1×10-8,1×10-9,1×10-10,0mmol/L尾加压素,每组再随机分成3小组,分别加入3H-Leu,3H-TdR,3H-UR3.7×1010Bq/mL。培养7d,分别于第1、4、7天测定样品3H-Leu,3H-TdR,3H-UR的放射性,并分别代表蛋白质、RNA、DNA的合成状态。细胞蛋白含量测定:将细胞培养于8孔塑料培养板中,随机分成4组,培养至第3天分别加入1×10-8,1×10-9,1×10-10,0mmol/L尾加压素,继续培养4d,收集细胞。Lowery法常规测细胞蛋白含量。结果:①在RPMI1640+300mL/L自体血清中,细胞生长、成熟、分裂状态良好,细胞合成DNA、RNA、蛋白质迅速,尤以3H-UR掺入作用较强。②成熟的人单核巨噬细胞与1×10-8,1×10-9,1×10-10,0mmol/L浓度范围的尾加压素孵育24h,未见细胞有形态学改变。1×10-10mmol/L浓度尾加压素在24h时即对3H-UR,3H-TdR掺入有抑制作用,随浓度升高和时间延长,作用均加强;且在1×10-9,1×10-8mmol/L浓度下对RNA作用较DNA强(0.608±0.005,0.730±0.007,t=18.8,P<0.01;0.500±0.007,0.571±0.004,t=10.4,P<0.01)。③加入1×10-8,1×10-9mmol/L浓度尾加压素与未加入尾加压素组比较对蛋白质的合成有抑制作用[(22.4±0.707)×10-3,(32.5±0.634)×10-3dpm,t=23.8,P<0.01;(27.35±0.636)×10-3,(32.5±0.634)×10-3dpm,t=12.8,P<0.01],但对蛋白质合成的抑制作用明显低于对DNA,RNA合成的抑制作用(0.688±0.003,0.571±0.004,0.500±0.007,t=12.3,17.5,P<0.001;0.840±0.007,0.730±0.007,0.608±0.005,t=9.6,31.2,P<0.001)。④1×10-8mmol/L尾加压素与成熟人单核巨噬AIM： To study the effects of urotensin, one of the optimal mini-vessel bioactive peptide that found recently on DNA, RNA and protein synthesis of human monocyte macrophages that cultured in vitro. METHODS： The experiment was conducted at the laboratory of Department of Cardiology, Southern Medical University and central laboratory of General Hospital of Guangzhou Military Area Command from October to December 2004. The 50 mL fresh venous blood was gained from healthy volunteers. The monocytes were separated with silica gel granule liquor （d= 1.130） coated with poly-acetamide pyrrolidine. Using the 12 holes plastic culture plate, the 0.5 mL cells at the concentration of 2×10^5 each per mL that had macrophage character in every hole, and divided randomly into 4 groups,1×10^-8 mmol/L, 1×10^-9 mmol/L, 1×10^-10 mmol/L,0 mmol/L urotensin were added, respectively, and the groups were assigned randomly into 3 subgroups, adding with 3H-Leu,3H-TdR,3H-UR 3.7×10^10 Bq/mL, respectively, cultured for 7 days. The radioactivities of 3H-Leu, 3H-TdR, 3H-UR Were detected at the 1st, 4th and 7th days, representing the synthesis status of protein, RNA and DNA, respectively. The detection of protein content： The cells were cultured in the 8-hole plastic cultured board, and assigned randomly into 4 groups. After culturing at the 3^rd day, the 1×10^-8 mmol/I, 1×10^-9 retool/L, 1×10^-10 retool/L, 0 mmol/L urotensin were added, respectively, cultured for 4 days continuously, then the cells were collected. The content of cell protein was detected routinely with the Lowery method. RESULTS： ① In the RPMI 1640＋300 mL/L autobody serum, the growth, mature and splitting status of cells were good. The synthesis of DNA, RNA and protein was rapid; especially the filter effect of 3H-UR was strong.② There was no change of cell morphology of mature human monocyte macrophage and urotensin at the concentration of 1×10^-8 mmol/L, 1×10^-9 mmol/L, 1×10^-10 mmol/L and 0 mmol/L incubated for 24 hours. The urotensin at the concent

关 键 词：血管升压素类/分析 血管升压素类/代谢 单核细胞 巨噬细胞 细胞 培养的 

分 类 号：R342[医药卫生—基础医学]