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目的探讨氟化钠(NaF)对大鼠成骨骨肉瘤UMR-106细胞及小鼠MC3T3-E1成骨细胞内Gs/Gi-AC-PKA信号途径的影响。方法以不同剂量NaF染毒UMR-106和MC3T3-E1细胞,培养24 h,用CCK-8法测定细胞增殖活性;用qPCR和Western Blot分别测定刺激性G蛋白(Stimulating adenylate cyclase g protein,Gs)、抑制性G蛋白(inhibitory adenylate cyclase g protein,Gi)、腺苷酸环化酶(adenylate cyclase,AC)和蛋白激酶A(protein kinase A,PKA)的mRNA和蛋白的表达水平。结果 UMR-106细胞和MC3T3-E1细胞NaF染毒后的存活率随NaF浓度增加而降低(P<0.05);不同剂量组之间,UMR-106细胞和MC3T3-E1细胞Gs、Gi mRNA的表达差异有统计学意义(P<0.05),且随着染毒剂量的升高呈先升高后下降趋势;AC mRNA表达及蛋白表达差异均有统计学意义(P<0.05),且呈明显下调趋势;UMR-106细胞和MC3T3-E1细胞的PKA mRNA和蛋白的表达差异有统计学意义(P<0.05),且随着染毒剂量的增加而呈明显上升趋势。结论 Gs/Gi-AC-PKA信号途径在氟骨症的形成过程中可能发挥一定作用。
Objective To investigate the effects of sodium fluoride (NaF) on Gs / Gi-AC-PKA signal pathway in osteogenic osteosarcoma UMR-106 cells and mouse MC3T3-E1 osteoblasts. Methods UMR-106 and MC3T3-E1 cells were exposed to different doses of NaF and cultured for 24 h. Cell proliferation was measured by CCK-8 assay. Stimulating adenylate cyclase g protein (Gs) , Inhibitory adenylate cyclase protein (Gi), adenylate cyclase (AC) and protein kinase A (PKA) mRNA and protein expression levels were detected. Results The survival rates of UMR-106 cells and MC3T3-E1 cells decreased with the increase of NaF concentration (P <0.05). The expressions of Gs and Gi mRNA in UMR-106 cells and MC3T3-E1 cells The difference was statistically significant (P <0.05), and then increased with the increase of dose, then decreased; AC mRNA expression and protein expression differences were statistically significant (P <0.05), and showed a significant downward trend; The expressions of PKA mRNA and protein in UMR-106 cells and MC3T3-E1 cells were significantly different (P <0.05), and increased with the increase of dose. Conclusion Gs / Gi-AC-PKA signaling pathway may play a role in the formation of skeletal fluorosis.