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目的研究斯钙素1(stanniocalcin 1,STC1)对内皮祖细胞(endothelial precusor cells,EPCs)增殖和迁移的影响并探讨其机制。方法密度梯度离心法分离、培养后,采用荧光双染法鉴定SD大鼠脾源性EPCs。不同浓度人重组斯钙素1(recombinant human stanniocalcin 1,rhSTC1)处理EPCs,结合siRNA技术干扰EPCs STC1表达,检测EPCs增殖能力及迁移能力的变化;观察STC1对蛋白激酶B(protein kinase B,PKB/Akt)、内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)及其磷酸化水平的影响;然后观察使用磷脂酰肌醇3激酶(phosphatidylinositol 3-kinase,PI3K)特异性抑制剂LY294002和eNOS特异性抑制剂L-NAME预处理的EPCs对rhSTC1的反应。结果荧光双染法鉴定EPCs阳性率约为88.5%;外源性给予rhSTC1呈浓度依赖性的促进EPCs增殖与迁移;而干扰内源性STC1的表达,则抑制其增殖与迁移;外源性给予rhSTC1增加EPCs中Akt和eNOS的磷酸化水平,而运用PI3K抑制剂(LY294002)和eNOS抑制剂(L-NAME)分别处理EPCs后,rhSTC1诱导的EPCs增殖与迁移能力明显降低。结论 STC1通过激活PI3K/Akt/eNOS信号通路,增强EPCs增殖、迁移能力,为损伤血管的修复提供了新的靶点。
Objective To investigate the effect of stanniocalcin 1 (STC1) on the proliferation and migration of endothelial progenitor cells (EPCs) and to explore its mechanism. Methods After density gradient centrifugation and culture, the splenic EPCs of SD rats were identified by fluorescence double staining. EPCs were treated with different concentrations of recombinant human stanniocalcin 1 (rhSTC1), and siRNA was used to interfere with the expression of STC1 in EPCs. The changes of proliferation and migration of EPCs were detected. The effect of STC1 on PKB / Akt, eNOS and eNOS were detected by immunohistochemistry. Then we observed the effect of phosphorylation of PI3K on the expression of eNOS and phosphorylation of phosphatidylinositol 3-kinase (PI3K) Specific inhibitor of L-NAME pretreatment of EPCs response to rhSTC1. Results The positive rate of EPCs identified by fluorescent double staining was about 88.5%. Exogenous rhSTC1 promoted the proliferation and migration of EPCs in a concentration-dependent manner, while inhibited the proliferation and migration of EPCs by interfering with the expression of endogenous STC1. rhSTC1 increased the phosphorylation levels of Akt and eNOS in EPCs, while EPCs treated with PI3K inhibitor (LY294002) and eNOS inhibitor (L-NAME) significantly reduced the proliferation and migration of EPCs induced by rhSTC1. Conclusion STC1 can enhance the proliferation and migration of EPCs by activating the PI3K / Akt / eNOS signaling pathway and provide a new target for the repair of damaged blood vessels.