Golpour Monireh, Fattahi Sadegh, Niaki Haleh Akhavan, Hadipoor Abbas, Abedian Zeinab, Ahangarian Gholam Reza, Parsian Hadi, Mosapour Abbas, Khorasani Hamid Reza, Vaziri Hamid Reza, Bijani Ali, Mostafazadeh Amrollah
Cellular and Molecular Biology Research Center, Babol University of Medical Sciences, Babol, Iran; University of Guilan, Rasht, Iran.
Biol Cell. 2014 May;106(5):139-50. doi: 10.1111/boc.201300063. Epub 2014 Apr 14.
There are several reports indicating that starved fibroblasts show higher proliferation rates when re-fed with foetal bovine serum. We have evidence demonstrating that this phenomenon is related to secretory proteins which may be beneficial to wound healing.
After re-feeding, 16 and 72 h serum-starved fibroblasts showed the highest and lowest proliferation rates, 1.59 and 0.51-fold difference compared to the non-starved control, respectively (P < 0.05). However, the latest value could be normalised by incubating cells with 16 h-starved fibroblast cell culture supernatant (16-SFS), prior to re-feeding. A strong correlation was found between total protein level in starved fibroblast culture supernatants and post re-feeding proliferation rates (r(2) = 0.90, P < 0.001). Two-dimensional gel electrophoresis analysis of 16-SFS confirmed the presence of proteins with relative molecular weights of 10-120 kDa and pI ranging from 4 to 6. A significant difference in calcium influx course was found between 16-SFS and the negative control (Dulbecco's Modified Eagle Medium) (P < 0.05). There was no significant difference in Ca(2+) concentrations after 1 h between non-starved controls and 16-SFS-treated fibroblasts. The scratch test demonstrated that the 16-SFS is able to induce fibroblast migration.
We concluded that human starved fibroblasts secrete proteins that are able to induce post re-feeding cell proliferation and fibroblasts migration, probably through the induction of a sustained calcium influx. This is worth being considered as a potential tool for wound healing.
有几份报告表明，饥饿的成纤维细胞在用胎牛血清重新喂养时显示出更高的增殖率。我们有证据表明，这种现象与可能对伤口愈合有益的分泌蛋白有关。
重新喂养后，血清饥饿16小时和72小时的成纤维细胞显示出最高和最低的增殖率，与未饥饿的对照组相比，分别有1.59倍和0.51倍的差异（P < 0.05）。然而，在重新喂养之前，用饥饿16小时的成纤维细胞培养上清液（16-SFS）孵育细胞可以使最新值正常化。在饥饿的成纤维细胞培养上清液中的总蛋白水平与重新喂养后的增殖率之间发现了很强的相关性（r(2) = 0.90，P < 0.001）。对16-SFS进行二维凝胶电泳分析证实存在相对分子质量为10 - 120 kDa且pI范围为4至6的蛋白质。在16-SFS和阴性对照（杜尔贝科改良伊格尔培养基）之间发现钙内流过程存在显著差异（P < 0.05）。未饥饿的对照组和16-SFS处理的成纤维细胞在1小时后Ca(2+)浓度没有显著差异。划痕试验表明16-SFS能够诱导成纤维细胞迁移。
我们得出结论，人类饥饿的成纤维细胞分泌的蛋白质可能通过诱导持续的钙内流来诱导重新喂养后的细胞增殖和成纤维细胞迁移。这值得被视为一种潜在的伤口愈合工具。
