奥曲肽对缺氧环境下大鼠胰腺外分泌细胞增殖、氧耗及HIF-1α、VEGF表达的影响

Effects of octreotide on rat pancreatic exocrine cell proliferation,oxygen consumption and ex⁃pression of HIF-1αand VEGF under hypoxic environment

目的:探究奥曲肽(Oct)在胰岛移植早期缺氧环境下对胰腺外分泌细胞AR42J的影响及可能机制。方法:将胰岛β细胞Min6分为常氧培养组和缺氧培养组,CCK-8法检测Min6细胞在常氧、缺氧环境中的增殖活性以确定培养时间。AR42J细胞分为常氧阴性对照组、常氧+Oct组、缺氧阴性对照组、缺氧+Oct组,将Oct稀释为不同浓度(10-6 mol/L、10-7 mol/L、10-8 mol/L、10-9 mol/L、10-10 mol/L)处理细胞。CCK-8法检测常氧、缺氧条件下Oct对AR42J增殖活性的影响,酶联免疫吸附试验(ELISA)法检测AR42J细胞培养上清液中血管内皮生长因子(VEGF)含量,磷氧探针检测细胞耗氧量,Western blotting法检测缺氧诱导因子(HIF-1α)蛋白表达。结果:Min6细胞在培养9 h后增殖活性达到最佳。与缺氧阴性对照组比较,缺氧+Oct组在作用3 h、6 h、9 h细胞活力上升(P<0.05)。与缺氧阴性对照组比较,除10-10 mol/L Oct组细胞HIF-1α表达无明显差异(P>0.05)外,其他各浓度Oct组HIF-1α蛋白表达量显著下降(均P<0.05)。与常氧阴性对照组比较,各浓度Oct组HIF-1α蛋白表达量均显著降低(均P<0.05)。与缺氧阴性对照组比较,缺氧+Oct组细胞上清VEGF含量和耗氧量降低(均P<0.05)。与常氧阴性对照组比较,各加药组HIF-1α表达降低(P<0.05),而VEGF含量、细胞耗氧量无明显差异(P>0.05)。结论:Oct提高缺氧环境下胰腺外分泌细胞活力,抑制HIF-1α与VEGF,降低细胞耗氧量,这可能是Oct对胰岛移植后缺氧环境中的胰岛细胞起到保护作用的机制之一。

Objective:To investigate the protective effect and possible mechanism of octreotide(Oct)on pancreatic exocrine Ar42Jcells in early hypoxic environment at the early stage of islet transplantation.Methods:Pancreaticβcells(Min6)cells were divided into normoxic culture group and hypoxic culture group.The CCK-8 method detected the proliferation activity of Min6 in normoxic and hypoxic environments to determine the culture time.AR42J cells were divided into normoxia negative control group,normoxia+Oct group,hypoxia negative control group,hypoxia+Oct group,and Oct was diluted into different concentrations(10-6,10-7,10-8,10-9,10-10mmol/L).CCK-8 method was used to detect the effect of Octon the proliferation activity of AR42J under normal and hypoxic conditions.The expressions of HIF-1αand VEGFproteins were detected by western blot and enzyme-linked immunosorbent assay(ELISA)respectively,moreover,the cell oxygen consumption was detected by phosphate oxygen probe.Results:The proliferation activity of Min6 reached the best after 9 h of culture.Compared with the hypoxia negative control group,the cell proliferation increased in the hypoxia+OCT group at 3 h,6 h,and 9 h(P<0.05).There was no significant difference in the expression of HIF-1αin the 10-10 mol/L Oct group(P>0.05),while the expression of HIF-1αin the other drug-added groups decreased(P<0.05);compared with the hypoxia negative control group,in thehypoxia+Oct group,VEGF expression and oxygen consumptiondecreased(P<0.05).Compared with the normoxia negative control group,the expression of HIF-1αin each addition group decreased(P<0.05),but in the normoxia+Oct group,there was no significant difference inVEGF and cell oxygen consumption(P>0.05).Conclusion:Oct can enhance the proliferation activity and reduce the expressions ofHIF-1α,VEGF and oxygen consumption of pancreatic exocrine cells in hypoxia.