钴原卟啉诱导大鼠骨髓间充质干细胞血红素加氧酶-1过表达及其对IL-10分泌的影响  

Effect of Cobalt Protoporphyrin on Hyperexpression of Heme Oxygenase-1 and Secretion of IL-10 in Rat Bone Marrow Mesenchymal Stem Cells

作　　者：韩夏[1] 白海[1] 王存邦[1] 高婧[2] 胡晓燕[1] 艾昊[1] 陈哲[1] 

机构地区：[1]兰州军区总医院血液科全军血液病中心,甘肃兰州730050 [2]甘肃省人民医院呼吸科,甘肃兰州730000

出　　处：《中国实验血液学杂志》2010年第5期1297-1301,共5页Journal of Experimental Hematology

摘　　要：本研究探讨不同浓度钴原卟啉(CoPP)诱导大鼠骨髓间充质干细胞(BMMSC)血红素加氧酶-1(hemeoxygenase-1,HO-1)过表达的效应关系及其对BMMSC分泌IL-10的影响。分离培养BMMSC,用流式细胞术检测BMMSC中CD34、CD45、CD44和CD71的表达;应用茜素红染色和油红染色检测BMMSC向成骨及成脂诱导情况。BMMSC随机分为4组,分别置于CoPP浓度为0、25、50和75μmol/L的培养液中进行诱导。用MTT法分析BMMSC增殖情况。采用RT-PCR检测各试验组HO-1的表达水平,筛选出诱导HO-1表达上调的最适合的CoPP浓度。用ELISA试剂盒检测各组IL-10的分泌量。结果表明,体外分离培养的BMMSC中CD34、CD45表达呈阴性,CD44和CD71表达呈阳性;BMMSC经成骨及成脂诱导后出现钙沉积和脂滴,茜素红染色和油红染色呈阳性。各个浓度的CoPP对细胞增殖无显影响,CoPP 50μmol/L能强烈诱导BMMSC的HO-1表达,其在试验组的表达高于对照组,与此同时IL-10分泌量也较对照组增多。结论:CoPP对细胞增殖活性无促进作用,CoPP50μmol/L是诱导BMMSC HO-1表达上调的最适合的浓度,HO-1表达上调可诱导IL-10分泌增加。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 adiopocyte 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.

关 键 词：钴原卟啉 骨髓间充质干细胞 血红素加氧酶-1 IL-10 

分 类 号：R979.5[医药卫生—药品]