尾加压素Ⅱ促NRK-49F细胞增殖及ECM分泌作用

目的探讨尾加压素Ⅱ（UrotensinⅡ,UⅡ）促进大鼠肾脏成纤维细胞（NRK-49F）增殖及细胞外基质（extracel-lular matrix,ECM）作用。方法分别用含不同浓度UⅡ（10-7、10-8、10-9和10-10mol.L-1）的培养液培养NRK-49F,采用MTT法检测细胞增殖情况,ELISA法检测细胞培养上清中Ⅰ型胶原（collagenⅠ,colⅠ）、Ⅲ型胶原（collagenⅢ,colⅢ）及纤连蛋白（fibronectin,FN）含量;RT-PCR法评价UⅡ抗体、尼莫地平和EDTA对UⅡ诱导NRK-49F colⅠ、colⅢ和FNmRNA过表达的抑制作用。结果 UⅡ可明显促进NRK-49F的增殖,且随着UⅡ浓度的增加而增强;10-9和10-8mol.L-1UⅡ可明显刺激NRK-49F ECM分泌,与对照组比较,差异显著（P〈0.01）;培养液中加入UⅡ抗体、尼莫地平和ED-TA后,NRK-49F colⅠ、colⅢmRNA表达量明显降低,与UⅡ组比较,有统计学意义（P〈0.01）。结论 UⅡ对体外培养的肾间质成纤维细胞有明显的促增殖作用,并且促进其ECM分泌,其机制可能与细胞外Ca2＋内流增加有关。

Role of Increasing Productivity and ECM Excreation in NRK-49F by Urotensin II

Objective To investigate the effects of urotensinⅡ（UⅡ） on the proliferation of the NRK-49F cells and explore the mechanism.Methods Various concentration of U II（10-7、10-8、10-9和10-10 mol·L-1）was added into the culture medium of NRK-49F,then the proliferation of NRK-49F was evaluated with MTT and the excretion of collagen I（col I）,collagen III（col III） and fibronectin（FN） was detected with ELISA.RT-PCR was applied to evaluate the mRNA expression of col I,col III and FN of NRK-49F treated with the antibody of U II,Nimodipine and EDTA.Results The proliferation of NRK-49F can be accelerated by UⅡ which shows dose dependent effects.When the antibody of UⅡ,Nimodipine and EDTA were added,the expression of col I and col III mRNA decreased significantly.Conclusion UⅡ can obviously accelerate the proliferation of NRK-49F and increase the ECM excreted by NRK-49F which maybe has relationship with the concentration of extracellular calcium.