【摘要】 为了研究rhALR在促进HepG2细胞增殖过程中对细胞Na+,K+ATPase的影响,采用四甲基噻唑盐(MTT)法检验了重组人肝再生增强因子(rhALR)对HepG2细胞的增殖作用并用无机磷比色法测定了细胞Na+,K+ATPase的酶活力。结果显示:rhALR能以浓度依赖的方式促进HepG2的细胞增殖,对细胞Na+,K+ATPase的酶活呈剂量时间依赖型影响;rhALR提高HepG2细胞Na+,K+ATPase的酶活符合MichaelisMenten方程作用机制,Vmax由0.84±0.11μmol·mg-1·min-1上升到1.68±0.07μmol·mg-1·min-1(p<0.01,差异有极显著性),而Km则由23.54±0.12mM变为20.86±0.13mM(p>0.05,差异无显著性),rhALR提高了细胞Na+,K+ATPase的转化效率;Na+,K+ATPase的专一性抑制剂(奎巴因)可以抑制rhALR促进HepG2细胞增殖的作用;ALR能以浓度时间依赖的方式激活细胞的Na+,K+ATPase,ALR引发的细胞增殖与了Na+,K+ATPase的活化有关。该为进一步研究rhALR的生物学功能提供了理论依据。更多还原

【Abstract】 In order to elucidate the effect on Na+, K+-ATPase in proliferation of HepG2 cell caused by recombinant human augmenter of liver regeneration (rh-ALR). MTT assay was used to measure the effect of rh-ALR on the proliferation of HepG2 cell, and the activities of the Na+, K+-ATPase were measured by inorganic phosphorus spectrophotometer. The result showed that rh-ALR could promote the proliferation of HepG2 cell in vitro, and rh-ALR stimulated the activation of Na+, K+-ATPase on HepG2 cells by concentration-time depended method. The mechanism of the stimulation in Na+, K+-ATPase on HepG2cell caused by rh-ALR was fit for a Michaelis-Menten equation model. The rh-ALR promoted the turnover rate of Na+, K+-ATPase, the V_ max increased from 0.84±0.11 μmol·mg -1·min -1 to 1.68±0.07 μmol·mg -1·min -1(p<0.01), K_m from 23.54±0.12 mM to 20.86±0.13 mM (p>0.05). The proliferation of HepG2 which caused by rh-ALR was inhibited by the Na+, K+-ATPase specific inhibitor, ouabain. This article presented that rh-ALR could stimulate the activation of Na+, K+-ATPase on HepG2 cells by concentration-time depended method. The proliferation of HepG2 cell caused by rh-ALR related with the activation of Na+, K+-ATPase. These results would be helpful in further research toward investigating the mechanisms of rh-ALR biological functions as the newly genetic engineering medicine.更多还原