大鼠骨髓间充质干细胞血红素加氧酶-1过表达对其IL-10分泌影响研究

发布时间：2018-01-15 10:01

本文关键词：大鼠骨髓间充质干细胞血红素加氧酶-1过表达对其IL-10分泌影响研究　出处：《兰州大学》2011年硕士论文　论文类型：学位论文

【摘要】：目的：本研究探讨不同浓度钴原卟啉(cobalt protoporphyrin, CoPP)诱导大鼠骨髓间充质干细胞(BMMSC)血红素加氧酶-1(heme oxygenase-1, HO-1)过表达的效应关系及对BMMSC分泌IL-10的影响。方法：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ELSIA试剂盒检测诱导组与实验组IL-10、TGFβ1和[FN-γ的分泌量。结果：流式细胞技术检测体外分离培养的BMMSC表面抗原结果显示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对细胞增殖无明显影响,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。诱导后BMMSC IL-10分泌量较对照组增高,TGFβi和IFN-γ的分泌量无明显变化。结论：CoPP对细胞增殖活性无明显影响；50μmol/L的CoPP是诱导BMMSC的HO-1表达上调的最适合的浓度；HO-1表达上调可诱导BMMSC IL-10分泌增加。
[Abstract]:Objective: to study cobalt protoporphyrin with different concentrations of cobalt protoporphyrin. CoPP- induced rat bone marrow mesenchymal stem cells (BMMSC) heme oxygenase-1 (heme oxygenase-1). The effect relationship of HO-1) overexpression and its effect on the secretion of IL-10 by BMMSC. Methods: BMMSC was isolated and cultured. The expression of CD34, CD45, CD44 and CD71 in BMMSC was detected by flow cytometry. Alizarin red staining and oil red staining were used to detect the osteogenesis and adipogenic induction of BMMSC. BMMSC was randomly divided into 4 groups with CoPP concentration of 0 ~ 25. The proliferation of BMMSC was analyzed by MTT method. The expression of HO-1 was detected by RT-PCR. The most suitable concentration of COPP to induce up-regulation of HO-1 expression was selected. IL-10 TGF- 尾 1 and TGF- 尾 1 were detected by ELSIA kit. [The secretion of FN- 纬. Results: flow cytometry was used to detect the surface antigens of BMMSC isolated in vitro. The results showed that the expression of CD34 + CD45 was negative and the expression of CD44 and CD71 was positive. Calcium deposition and lipid droplet were observed after osteogenesis and lipogenesis of BMMSC. Alizarin red staining and oil red staining were positive. Different concentrations of CoPP had no effect on cell proliferation. CoPP 50 渭 mol/L could strongly induce the HO-1 expression of BMMSC, and its expression in the experimental group was higher than that in the control group, and the secretion of BMMSC IL-10 was higher than that in the control group. The secretion of TGF 尾 I and IFN- 纬 did not change significantly. Conclusion% Copp has no significant effect on cell proliferation activity. 50 渭 mol/L CoPP was the most suitable concentration to induce the up-regulation of HO-1 expression in BMMSC. Upregulation of HO-1 expression can induce the increase of BMMSC IL-10 secretion.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R329

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