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目的:探讨ghrelin对大鼠心肌微血管内皮细胞(cardiac microvascular endothelial cells,CMECs)增殖、迁移能力和分泌NO的影响。方法:以酶消化法获得SD大鼠的CMECs,采用Dil-ac-LDL吞噬试验鉴定CMECs。将新一代CMECs分5组分别干预24 h:即(A)空白对照组、(B)1×10-9mol/L ghrelin组、(C)1×10-8mol/L ghrelin组、(D)1×10-7mol/L ghrelin组和(E)ghrelin+拮抗剂LY294002组(1×10-7mol/L ghrelin和20μmol/L LY294002)。用MTT比色法检测ghrelin对CMECs增殖的影响;以transwell法检测ghrelin对CMECs迁移的影响。用硝酸还原法检测CMECs分泌NO的量。结果:Dil-ac-LDL吞噬试验表明,90%以上的CMECs表达Dil-ac-LDL。与对照组相比,10-8~10-7mol/L的ghrelin可明显促进CMECs增殖和迁移,以及NO分泌(P<0.05);LY294002则能够明显抑制ghrelin对CMECs的促增殖和迁移作用,以及抑制ghrelin刺激CMECs分泌NO的作用(P>0.05)。结论:一定浓度条件下,ghrelin能够促进心肌微血管内皮细胞(CMEC)的增殖、迁移和NO分泌,这可能与激活AKT/PI3K信号转导通路有关。
Objective: To investigate the effect of ghrelin on proliferation, migration and secretion of NO in rat cardiac microvascular endothelial cells (CMECs). Methods: CMECs of SD rats were obtained by enzymatic digestion. The Dil-ac-LDL phagocytosis assay was used to identify CMECs. The new generation of CMECs were divided into 5 groups for 24 h: (A) blank control group, (B) 1 × 10-9 mol / L ghrelin group, (C) 1 × 10-8 mol / L ghrelin group, × 10-7mol / L ghrelin group and (E) ghrelin + antagonist LY294002 group (1 × 10-7mol / L ghrelin and 20μmol / L LY294002). The effect of ghrelin on the proliferation of CMECs was detected by MTT colorimetric assay. The effect of ghrelin on the migration of CMECs was detected by transwell assay. Nitric acid reduction was used to detect the amount of NO secreted by CMECs. Results: Dil-ac-LDL phagocytosis test showed that more than 90% of CMECs expressed Dil-ac-LDL. Compared with the control group, ghrelin of 10-8-10-7mol / L significantly promoted the proliferation and migration of CMECs and NO secretion (P <0.05); LY294002 significantly inhibited the proliferation and migration of CMECs induced by ghrelin, and Inhibition of ghrelin stimulated the secretion of NO in CMECs (P> 0.05). CONCLUSION: Ghrelin can promote the proliferation, migration and NO secretion of myocardial microvascular endothelial cells (CMEC) under certain concentration, which may be related to the activation of AKT / PI3K signal transduction pathway.