摘要
目的探讨尾加压素Ⅱ（UⅡ）对大鼠近端肾小管上皮NRK-52E细胞增殖和细胞周期的影响及其机制。方法在NRK-52E细胞培养液中加入10^-10、10^-9、10^-8和10^-7mol/L UⅡ,同时设UⅡ活性阻断组：UⅡ＋尼卡地平和UⅡ＋EDTA,以单纯DMEM为对照组。培养48 h后,采用5-溴-2′-脱氧尿嘧啶核苷（BrdU）掺入法检测各组细胞的增殖活性,流式细胞术分析细胞周期。结果UⅡ（10^-10-10^-8mol/L）以浓度依赖方式促进NRK-52E细胞BrdU掺入（A值分别为0.491±0.038、0.291±0.024和0.281±0.037）,其中以10-8mol/L UⅡ的作用最明显,10^-7mol/L UⅡ对NRK-52E细胞增殖的影响与对照组比较差异无显著意义。UⅡ影响NRK-52E细胞周期,在10^-10-10^-8mol/L浓度范围内增加S期细胞百分比（分别为26.96%±3.35%、44.26%±3.28%、48.12%±2.22%）。尼卡地平和EDTA均可以降低UⅡ诱导NRK-52E细胞的BrdU掺入增加,降低S期细胞百分比。结论UⅡ具有较强促进NRK-52E细胞增殖的作用,但大剂量则无此作用,其诱导NRK-52E细胞增殖作用部分是通过Ca^2＋内流来介导的。
Objective To explore the effect of urotensin Ⅱ（U Ⅱ ） on proliferation and cell cycle of rat proximal tubular epithelium NRK-52 cells and its mechanism.Methods Divide NRK-52E cells into one control and six test groups. The cells in the six test grops were incubated for 48 h after addition of 10^-10,10^- 9,10^ - 8 and 10 - 7moL/L U Ⅱ , 10 - 5 mol/L nicardipine plus 10- Smol/L U Ⅱ and 2 × 10^-3mol/L EDTA plus 10^-8 mol/L U Ⅱ respectively.However, no any stimulant was added in the cells in control group.Determine the proliferation activity of cells in various groups by BrdU incorporation method and the cell cycle by flow cytometry. Results At the concentration range of 10^-10 - 10^-8mol/L, U Ⅱ showed a concentration-dependent promoting effect on the incorporation of BrdU into NRK-52E cells. The incorporation （A values） of BrdU at U Ⅱ concentrations of 10 ^- 10,10^ - 9 and 10^- 8mol/L were 0.491 ± 0.038,0.291 ± 0.024 and 0.281 ± 0.037 respectively. However, 10-Tmol/L U Ⅱ showed no significant effect on the proliferation of NRK-52E cells.After treatment with 10^-10,10^-9 and 10^-8mol/L U Ⅱ ,the percentage of cells at S phase were （26.96 ± 3.35） %, （44.26 ± 3.28） % and （48.12 ± 2.22） % respectively. Either nicardipine or EDTA decreased the incorporation of BrdU induced by U Ⅱ and the pementage of cells at S phase. Conclusion The U Ⅱ at a certain cocentration promoted the proliferation of NRK-52E cells significantly. The induction of proliferation of NRK-52E cells was partially mediated by calcium ion inflow.
出处
《中国生物制品学杂志》
CAS
CSCD
2008年第2期99-102,共4页
Chinese Journal of Biologicals
基金
国家自然科学基金资助课题(30570855)
