血管紧张素Ⅱ对成骨细胞IL-6表达的影响  

Effects of angiotensinⅡon expression of IL-6 in osteoblasts

作　　者：徐良志 张晨[2] 

机构地区：[1]核工业四一七医院骨科,临潼710600 [2]西安交通大学医学院第二附属医院骨一科,西安710004

出　　处：《郑州大学学报（医学版）》2014年第5期629-633,共5页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金资助项目81301562;西安交通大学基本科研业务费学科综合交叉科研项目xjj2014152

摘　　要：目的:研究血管紧张素Ⅱ(AngⅡ)对成骨细胞MC3T3-E1 IL-6表达的影响。方法:取对数生长期的MC3T3-E1细胞,随机分为对照组、AngⅡ处理组、奥美沙坦+AngⅡ处理组、NAC+AngⅡ处理组以及PDTC+AngⅡ处理组。然后分别利用RT-PCR和ELISA的方法检测各组细胞中IL-6的表达,用荧光显微镜检测细胞内活性氧的变化,用Western blot方法测定NK-κB p65磷酸化水平,用细胞免疫荧光法检测NK-κB p65蛋白在细胞内的定位。结果:与对照组比较,AngⅡ处理组细胞中IL-6 mRNA和蛋白的表达均明显增加;与AngⅡ处理组比较,奥美沙坦、NAC和PDTC+AngⅡ组细胞中IL-6 mRNA和蛋白的表达降低(F=46.971和38.048,P<0.001)。与对照组比较,AngⅡ处理组细胞内活性氧水平明显增加;与AngⅡ处理组比较,奥美沙坦和NAC+AngⅡ组细胞内活性氧水平降低(F=57.413,P<0.001)。与对照组比较,AngⅡ增加了细胞中NK-κB p65的磷酸化水平;与AngⅡ处理组比较,奥美沙坦、NAC和PDTC+AngⅡ组细胞中NK-κB p65的磷酸化水平降低(F=44.511,P<0.001)。结论:AngⅡ通过其Ⅰ型受体促进成骨细胞内活性氧的产生,进而作用于NF-κB信号通路,最后促进成骨细胞高表达IL-6。Aim：To study the effects of Angiotensin Ⅱ （ Ang Ⅱ ） on IL-6 expression in osteoblasts in MC3T3-E1 cells. Methods：MC3T3-E1 cells.in log phase were randomly divided into control group, Ang Ⅱ group,olmesartan ＋ Ang Ⅱ group,NAC ＋ Ang Ⅱ group and PDTC ＋ Ang Ⅱ group. Expressions of IL-6 was determined by RT-PCR and ELISA ana- lyses. Intracellular reactive oxygen species （ROS） was measured with a fluorescence microscope and the phosphorylation level of NK-κB p65 was examined by Western-blot. Changes of NK-KB p65 protein in the nucleus was detected by immunofluorescence. Fesults：Expression of IL-6 in Ang I1 group was obviously increased than that in control group, however, olmesartan,NAC and PDTC decreased IL-6 expression more than that in Ang 11 group （ F = 46. 971 and 38. 048, P 〈 0. 001 ）. lntracellular ROS in MC3T3-E1 cells was obviously increased than that in control group, however,olmesartan, NAC and PDTC decreased ROS expression more than that in Ang Ⅱ group（ F = 57. 413 ,P 〈 0. 001 ） ; Phosphorylation of NK-κB p65 was obviously increased than that in control group, however, olmesartan, NAC and PDTC decreased Phosphorylation of NK-KB more than that in Ang Ⅱ group（ F = 44. 511 ,P 〈 0. 001 ）. Conclusion ：Type I receptor of Ang Ⅱ promotes active oxygen in osteoblasts,acts on NF- κB pathway,and finally promotes the high expression of IL-6 in osteoblasts.

关 键 词：血管紧张素Ⅱ 成骨细胞 IL-6 活性氧 NK—κB信号通路 

分 类 号：R322.7[医药卫生—人体解剖和组织胚胎学]