相对分子质量透明质酸在脐静脉内皮细胞增殖及连接蛋白Ezrin磷酸化表达中的作用研究

Effects of hyaluronic acid on human umbilical vein endothelial cells proliferation and expression of membrane-cytoskeleton linker protein Ezrin

目的探讨高相对分子质量透明质酸(nHA)和低相对分子质量透明质酸(oHA)对脐静脉内皮细胞增殖能力及膜-细胞骨架连接蛋白埃兹(Ezrin)表达的影响。方法分别用nHA和oHA处理人脐静脉内皮细胞,以5-溴脱氧尿嘧啶核苷(Brdu)酶联免疫吸附试验(ELISA)检测透明质酸(HA)作用后细胞增殖能力;用荧光实时定量聚合酶链反应(PCR)和蛋白免疫印迹试验(Western blot)检测HA对Ezrin mRNA和蛋白表达的影响。结果与对照组相比,72 h后oHA(10μg/mL)显著促进血管内皮细胞增殖(P<0.001),24 h后促进Ezrin基因的表达,48 h后促进Ezrin蛋白磷酸化表达。而nHA(200μg/mL)则抑制血管内皮细胞的增殖(P<0.01)和Ezrin蛋白的表达磷酸化。结论oHA能有效促进血管内皮的增殖并促进其胞内连接蛋白Ezrin的表达磷酸化,nHA则抑制内皮细胞的增殖和Ezrin蛋白的表达。

Objective To investigate the effects of native high molecular weight hyaluronic acid （nHA） and low molecular weight hyaluronic acid（oHA） on the proliferation of human umbilical vein endothelial cell and the expression of membrane-cytoskeleton linker protein Ezrin. Methods Cell proliferation ability was detected by 5-bromo 2-deoxyuridine （Brdu） enzyme-linked immunosorbent assay （ELISA）. Expression of Ezrin mRNA and protein was detected by real-time polymerase chain reaction（PCR） and Western blot after being treated with oHA and nHA in human umbilical vein endothelial cells. Results Human umbilical vein endothelial cell proliferation ability was higher than control group after being treated with oHA at the concentration of 10 μg/mL for 72 h（ P 〈0.001 ）. Ezrin mRNA expression was upregulated after being treated with oHA for 24 h. The expression of Ezrin protein , especially the expression of phosphorylated Ezrin was upregulated by oHA for 48 h. nHA （200 μg/mL）inhibited human umbilical vein endothelial cell proliferation （ P 〈 0.01 ） and Ezfin phosphorylated. Conclusions oHA may promote endothelial cell proliferation and the expression of membrane-cytoskeleton linker protein Ezrin. nHA inhibits cell proliferation and the expression of Ezrin.