Effect of cobalt protoporphyrin on hyperexpression of heme oxygenase-1 and secretion of IL-10 in rat bone marrow mesenchymal stem cells / 中国实验血液学杂志

ABSTRACT

This study was aimed to investigate the effects of cobalt protoporphyrin (CoPP) on hyperexpression of heme oxygenase-1 (HO-1) and secretion of IL-10 in bone marrow-derived mesenchymal stem cells (BMMSC). The BMMSC were isolated from rats and cultured, then were randomly divided into 4 groups, and were induced in culture medium with CoPP of 0, 25, 50, and 75 µmol/L respectively. The expressions of CD34, CD45, CD44 and CD71 in BMMSC were detected by flow cytometry; the osteoblast and adipocyte induction of BMMSC was determined by alizarin red and oil red staining respectively; the proliferation status of BMMSC was assayed by MTT method; the HO-1 expression in test groups was detected by RT-PCR; the optimal concentration of CoPP for induction of HO-1 expression was screened. The ELISA kit was used to detect the HO-1 levels in each groups. The results showed that the expressions of CD34, CD45 in BMMSC were negative while the expressions of CD44 and CD71 in BMMSC were positive. After induced by osteoblast and adipocyte inductor, BMMSC were positive for alizarin red staining and oil red staining. Various concentrations of CoPP had no significant impact on cell proliferation. When the CoPP concentration of 50 µmol/L could induce strong expression of HO-1, expression level in test groups was significantly higher than that in control group, meanwhile the IL-10 secretion was significantly higher than that in control group. It is concluded that CoPP has no ability to promote cell proliferation, 50 µmol/L is the optimum concentration for up-regulation of CoPP-induced BMMSC HO-1 expression, the up-regulation of HO-1 expression can increased IL-10 secretion significantly.