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目的观察Apelin对小鼠成骨细胞MC3T3-E1增殖和分化的作用,探讨其可能的作用机制。方法 RT-PCR检测MC3T3-E1细胞APJ(Apelin受体)的表达、Western blot、ELISA、放射免疫法和[3H]TdR-掺入法检测MAPKs和Akt的激活,以及MC3T3-E1细胞的增殖和分化指标;用PI3-K抑制剂LY294002,AKT抑制剂HIMO和JNK抑制剂SP600125预处理以观察Apelin影响MC3T3-E1细胞增殖的信号转导机制。结果 MC3T3-E1细胞表达APJ;Apelin对MC3T3-E1细胞碱性磷酸酶(alkali phosphatase,ALP)活性、骨钙素(osteocalcin,OC)和Ⅰ型胶原(type Ⅰ collagen)的分泌及Runx2蛋白的表达无影响;Apelin促进MC3T3-E1细胞增殖;Apelin诱导JNK和PI3-K/AKT的磷酸化;PI3-K抑制剂LY294002,AKT抑制剂HIMO和JNK抑制剂SP600125均可以抑制Apelin对MC3T3-E1细胞的促增殖作用。结论 MC3T3-E1细胞表达APJ;Apelin通过JNK和PI3-K信号转导通路促进MC3T3-E1细胞增殖,但对其分化无影响。
Objective To observe the effect of Apelin on proliferation and differentiation of mouse osteoblast MC3T3-E1 and to explore its possible mechanism. Methods The expression of APJ (Apelin receptor) in MC3T3-E1 cells was detected by RT-PCR. The activation of MAPKs and Akt were detected by Western blot, ELISA, radioimmunoassay and [3H] TdR- PI3-K inhibitor LY294002, AKT inhibitor HIMO and JNK inhibitor SP600125 were used to observe the signal transduction mechanism of Apelin on the proliferation of MC3T3-E1 cells. Results AP3 was expressed in MC3T3-E1 cells; alkaline phosphatase (ALP) activity, secretion of osteocalcin (OC) and type Ⅰ collagen and expression of Runx2 protein in MC3T3- Apelin could promote the proliferation of MC3T3-E1 cells; Apelin induced the phosphorylation of JNK and PI3-K / AKT; PI3-K inhibitor LY294002, AKT inhibitor HIMO and JNK inhibitor SP600125 could inhibit the effect of Apelin on MC3T3-E1 cells Promote the role of proliferation. Conclusion MC3T3-E1 cells express APJ. Apelin can promote the proliferation of MC3T3-E1 cells through JNK and PI3-K signal transduction pathways, but have no effect on the differentiation.