尾加压素Ⅱ促进乳鼠心肌成纤维细胞分泌胶原及增殖的细胞内信号转导机制

目的探讨尾加压素Ⅱ(UⅡ)促进乳鼠心肌成纤维细胞(CFs)分泌胶原及增殖的细胞内信号转导机制。方法体外培养CFs,采用免疫组织化学染色法及羟脯氨酸测定法分别观察不同浓度UⅡ作用下,CFs中磷酸化ERK1/2细胞灰度和CFs培养上清中胶原含量的变化;蛋白激酶C(PKC)抑制剂chelerythrine chloride(Che)、ERK1/2抑制剂PD98059和钙调神经磷酸酶(CaN)抑制剂cyclosporin A(CsA)各自对UⅡ诱导的细胞增殖的影响。结果在1×10-10、1×10-9、1×10-8mol/L UⅡ作用下,CFs培养上清中胶原含量均较对照组明显增加,而CFs中磷酸化ERK1/2细胞灰度均较对照组有显著降低(P<0.01);在1×10-7mol/L UⅡ作用下,上述各参数与对照组比较无统计学意义(P>0.05)。1×10-6mol/L Che+1×10-8mol/L UⅡ组、1×10-5mol/L PD98059+1×10-8mol/L UⅡ组和5μg/mLCsA+1×10-8mol/L UⅡ组的胶原含量均高于对照组而低于1×10-8mol/L UⅡ组(P<0.05)。1×10-6mol/L Che+1×10-8mol/L UⅡ组、1×10-5mol/L PD98059+1×10-8mol/L UⅡ组和5μg/mL CsA+1×10-8mol/L UⅡ组的p-ERK1/2的灰度低于对照组(P<0.01);1×10-6mol/L Che+1×10-8mol/L UⅡ组和5μg/mL CsA+1×10-8mol/LUⅡ组的p-ERK1/2的灰度高于1×10-8mol/L UⅡ组(P<0.01),而1×10-5mol/L PD98059+1×10-8mol/L UⅡ组与之比较则无统计学意义(P>0.05)。结论UⅡ具有促进CFs分泌胶原及增殖的作用,其作用可能是通过PKC/MAPK/CaN途径实现的。

Effect of urotensin Ⅱ on the proliferation of cardiac myofibroblasts and its intracellular signaling mechanism in new-born SD rats

Objective To explore the effect of urotensin Ⅱ(UⅡ) on the proliferation of cardiac fibroblasts(CFs) and its intracellular signaling mechanism in new-born SD rats.Methods Isolated and cultured CFs from new-born SD rats were used,and cell proliferation was observed by immunohistochemical staining and hydroxyproline assay.Results With the treatment of 10-10,and 10-9,and 10-8mol/L UⅡ,the collagen content in CFs supernatant was significantly higher than that in control group,the grey scale of p-ERK1/2 was significantly lower than that of control(P<0.01),while the indices in 10-7mol/L UⅡ group were not significantly different from those of control(P>0.05).Compared with that in control group,the collagen content in 10-6mol/L Che+10-8mol/L UⅡ and 5 μg/mL CsA+10-8 mol/L UⅡ groups increased(P<0.05),while no difference was found between 10-5mol/L PD98059+10-8mol/L UⅡ and control groups(P>0.05).Compared with those in control group,the grey scales of p-ERK1/2 in 10-6 mol/L Che+10-8mol/L UⅡ,and 10-5mol/L PD98059+10-8mol/L UⅡ,5 μg/mL CsA+10-8mol/L UⅡ groups declined significantly(P<0.01).In contrast to those in 10-8mol/L UⅡ group,the grey scales in 10-6mol/L Che+10-8mol/L UⅡ and 5 μg/mL CsA+10-8 mol/L UⅡ groups increased significantly(P<0.01),but had no difference for 10-5mol/L PD98059+10-8 mol/L UⅡ group(P>0.05).Conclusion UⅡ can stimulate the proliferation of CFs in new-born SD rats and the UⅡ-induced proliferation of CFs is probably mediated by PKC/MAPK/CaN.