本文關(guān)鍵詞：大鼠骨髓間充質(zhì)干細胞血紅素加氧酶-1過表達對其IL-10分泌影響研究　出處：《蘭州大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 鈷原卟啉 大鼠骨髓間充質(zhì)干細胞 血紅素加氧酶-1 IL-10

【摘要】：目的：本研究探討不同濃度鈷原卟啉(cobalt protoporphyrin, CoPP)誘導(dǎo)大鼠骨髓間充質(zhì)干細胞(BMMSC)血紅素加氧酶-1(heme oxygenase-1, HO-1)過表達的效應(yīng)關(guān)系及對BMMSC分泌IL-10的影響。 方法：分離培養(yǎng)BMMSC;用流式細胞術(shù)檢測BMMSC中CD34、CD45、CD44和CD71的表達；應(yīng)用茜素紅染色和油紅染色檢測BMMSC向成骨及成脂誘導(dǎo)情況。BMMSC隨機分為4組,分別置于CoPP濃度為0、25、50和75μmol/L的培養(yǎng)液中進行誘導(dǎo)。用MTT法分析BMMSC增殖情況。采用RT-PCR檢測各試驗組HO-1的表達水平,篩選出誘導(dǎo)HO-1表達上調(diào)的最適合的COPP濃度。用ELSIA試劑盒檢測誘導(dǎo)組與實驗組IL-10、TGFβ1和[FN-γ的分泌量。 結(jié)果：流式細胞技術(shù)檢測體外分離培養(yǎng)的BMMSC表面抗原結(jié)果顯示CD34、CD45表達呈陰性,CD44和CD71表達呈陽性；BMMSC經(jīng)成骨及成脂誘導(dǎo)后出現(xiàn)鈣沉積和脂滴,茜素紅染色和油紅染色呈陽性。各個濃度的CoPP對細胞增殖無明顯影響,CoPP 50μmol/L能強烈誘導(dǎo)BMMSC的HO-1表達,其在試驗組的表達高于對照組。誘導(dǎo)后BMMSC IL-10分泌量較對照組增高,TGFβi和IFN-γ的分泌量無明顯變化。 結(jié)論：CoPP對細胞增殖活性無明顯影響；50μmol/L的CoPP是誘導(dǎo)BMMSC的HO-1表達上調(diào)的最適合的濃度；HO-1表達上調(diào)可誘導(dǎo)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- 緯. 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- 緯 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.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R329

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