Effects of haem oxygenase-1 expression on oxidative injury and biological behaviours of rat dermal fibroblasts
Effects of haem oxygenase-1 expression on oxidative injury and biological behaviours of rat dermal fibroblasts
复制标题
血红素加氧酶1表达对大鼠真皮成纤维细胞氧化损伤及生物学行为的影响
DOI：
10.12968/jowc.2018.27.11.780
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发表时间：
2018-11-01
影响因子：
1.9
通讯作者：
Chen, Li-hong
中科院分区：
文献类型：
--
作者：
Li, Qing-ling;Guo, Ruo-mi;Chen, Li-hong
目的：本研究探讨血红素氧合酶-1（HO-1）高表达对高糖条件下大鼠真皮成纤维细胞氧化损伤及生物学行为的影响。胆红素试剂盒、实时荧光定量PCR和Western blotting分别检测HO-1的蛋白酶活性、mRNA和蛋白水平。酶联免疫吸附测定（ELISA）试剂盒测量8-羟基脱氧鸟苷（8-OHdG）、活性氧（ROS）和胶原（羟脯氨酸）分泌的培养基水平。使用流式细胞术测量细胞增殖。采用Hoechst 33258染色和流式细胞术检测细胞凋亡。结果：HO-1表达呈时间依赖性变化，高糖组HO-1表达在96 h时最低，正常葡萄糖组HO-1表达在96 h时最低。与NG组相比，HG组在96 h时8-OHdG、ROS和细胞凋亡增加，胶原分泌、细胞增殖和细胞迁移（水平和垂直方向）减少。与NG/HG组相比，NG/HG + NH/HG组和HH组HO-1表达持续升高至少96 h，细胞8-OHdG和ROS减少，胶原合成增加，增殖和迁移能力增强，凋亡减少。结论：Haemin诱导成纤维细胞HO-1表达，可能影响成纤维细胞的氧化损伤和生物学行为。这些发现表明HO-1可能通过减轻氧化损伤和改善成纤维细胞的生物学行为来加速糖尿病伤口的愈合。利益声明：作者没有利益冲突需要声明。
Objective: This study investigated the effects of high haem oxygenase-1 (HO-1) expression on oxidative injury and the biological behaviours of rat dermal fibroblasts, under high glucose conditions.Method: Rat dermal fibroblasts were cultured in normal glucose (1.0g/l), high glucose (4.5g/l) or haemin (5 mu m). A bilirubin kit, real-time polymerase chain reaction (RT-PCR) and Western blotting measured the protease activity, mRNA, and protein levels of HO-1, respectively. An enzyme-linked immunosorbent assay (ELISA) kit measured media levels of 8-hydroxydeoxyguanosine (8-OHdG), reactive oxygen species (ROS) and collagen (hydroxyproline) secretion. Cell proliferation was measured using flow cytometry. Cell apoptosis was measured using Hoechst 33258 staining and flow cytometry. The transwell method and scratch test evaluated cell migration.Results: HO-1 expression exhibited a time-dependent change that was lowest in the high glucose (HG) group at 96 hours compared with the normal glucose (NG) group. In the HG group, the 8-OHdG, ROS and cell apoptosis were increased, and collagen secretion, cell proliferation and cell migration (horizontal and vertical) were decreased compared with the NG group at 96 hours. Haemin treatment sustained high HO-1 expression for at least 96 hours, and the cells exhibited decreased 8-OHdG and ROS, increased collagen synthesis, improved proliferation and migration ability, and decreased apoptosis in the NG and haemin (NH) group/HG and haemin (HH) group compared with the NG/HG groups. These cells recovered from oxidative injury and biological behaviours dysfunction.Conclusion: Haemin induces HO-1 expression in fibroblasts and it may influence the oxidative injury and biological behaviours of fibroblasts. These findings suggest that HO-1 may accelerate the healing of diabetic wounds via alleviation of oxidative injury and improvement of biological behaviours of fibroblasts.Declaration of interest: The authors have no conflicts of interest to declare.