基質細胞衍生因子-1 α /CXCR4/CXCR7軸對骨髓間充質干細胞遷移的影響
發(fā)布時間:2021-07-30 22:47
目的探討基質細胞衍生因子-1α(SDF-1α)受體CXCR4、CXCR7在骨髓間充質干細胞(BMSCs)中蛋白和mRNA的表達;及SDF-1α/CXCR4/CXCR7軸對BMSCs遷移作用的可能機制。方法體外培養(yǎng)大鼠BMSCs,流式細胞術鑒定細胞表面抗原CD29、CD44和CD34。應用CXCR4特異性拮抗劑AMD3100及CXCR7中和抗體分別阻斷CXCR4及CXCR7,通過Western blotting和RT-PCR分別檢測BMSCs蛋白和mRNA的表達變化;Transwell法檢測細胞遷移能力。本次實驗分為單純BMSCs組(A)、AMD3100預處理BMSCs組(B)、CXCR7中和抗體預處理BMSCs組(C)及AMD3100+CXCR7中和抗體預處理BMSCs組(D)。結果經(jīng)鑒定第3代大鼠BMSCs中CD29和CD44均呈陽性表達,而CD34表達陰性。BMSCs中CXCR4、CXCR7蛋白和mRNA均有表達。與A組相比,B組及D組CXCR4及CXCR7蛋白表達明顯受到抑制(P<0.05),C組只有CXCR7蛋白表達降低(P<0.05);各組CXCR4 mRNA和C...
【文章來源】:解剖學報. 2014,45(05)北大核心CSCD
【文章頁數(shù)】:7 頁
【部分圖文】:
BMSCs的培養(yǎng)與鑒定A.第3代BMSCs;B~D分別為流式細胞術檢測第3代細胞CD44、CD34、CD29表達標尺示100μmFig.1CultureandidentificationofBMSCsA,ThethirdpassageofBMSCs;B-DistheexpressionofCD44,CD34,CD29ofthethirdpassagecellsbyflowcytometryrespectivelyBar=100μm
·642·解剖學報45卷,5期圖1BMSCs的培養(yǎng)與鑒定A.第3代BMSCs;B~D分別為流式細胞術檢測第3代細胞CD44、CD34、CD29表達標尺示100μmFig.1CultureandidentificationofBMSCsA,ThethirdpassageofBMSCs;B-DistheexpressionofCD44,CD34,CD29ofthethirdpassagecellsbyflowcytometryrespectivelyBar=100μm圖2RT-PCR檢測不同處理組細胞CXCR4mRNA、CXCR7mRNA的表達情況M.DNA分子量標準;A.單純BMSCs組;B.AMD3100組;C.CXCR7中和抗體組;D.AMD3100+CXCR7中和抗體組,不同處理組細胞與單純BMSCs組相比,mRNA表達無影響(x珋±s,n=3)Fig.2DetectionofCXCR4mRNA,CXCR7mRNAbyRT-PCRM,DNAmaker;A,BMSCsgroup;B,AMD3100group;C,CXCR7-specificneutralizingantibodygroup;D,AMD3100+CXCR7-specificneutralizingantibodygroup.ComparedwithBMSCsgroup,theexpressionofmRNAarenotchangedbydifferenttreatments(x珋±s,n=3)圖3Westernblotting檢測AMD3100及CXCR7中和抗體對BMSCs中CXCR4、CXCR7蛋白表達的影響A.單純BMSCs組;B.AMD3100組;C.CXCR7中和抗體組;D.AMD3100+CXCR7中和抗體組,與單純BMSCs組相比,*P<0.05,#P<0.05;與AMD3100組相比,▲P<0.05(x珋±s,n=3)Fig.3WesternblottinganalysisofCXCR4,CXCR7proteinexpressioninBMSCspretreatedwithAMD3100orCXCR7-specificneutralizingantibodyA,BMSCsgroup;B,AMD3100group;C,CXCR7-specificneutralizingantibodygroup;D,AMD3100+CXCR7-specificneutralizingantibodygroup;*P<0.05,#P<0.05,vsBMSCsgroup;▲P<0.05,vsAMD3100group(x珋±s,n=3)
·642·解剖學報45卷,5期圖1BMSCs的培養(yǎng)與鑒定A.第3代BMSCs;B~D分別為流式細胞術檢測第3代細胞CD44、CD34、CD29表達標尺示100μmFig.1CultureandidentificationofBMSCsA,ThethirdpassageofBMSCs;B-DistheexpressionofCD44,CD34,CD29ofthethirdpassagecellsbyflowcytometryrespectivelyBar=100μm圖2RT-PCR檢測不同處理組細胞CXCR4mRNA、CXCR7mRNA的表達情況M.DNA分子量標準;A.單純BMSCs組;B.AMD3100組;C.CXCR7中和抗體組;D.AMD3100+CXCR7中和抗體組,不同處理組細胞與單純BMSCs組相比,mRNA表達無影響(x珋±s,n=3)Fig.2DetectionofCXCR4mRNA,CXCR7mRNAbyRT-PCRM,DNAmaker;A,BMSCsgroup;B,AMD3100group;C,CXCR7-specificneutralizingantibodygroup;D,AMD3100+CXCR7-specificneutralizingantibodygroup.ComparedwithBMSCsgroup,theexpressionofmRNAarenotchangedbydifferenttreatments(x珋±s,n=3)圖3Westernblotting檢測AMD3100及CXCR7中和抗體對BMSCs中CXCR4、CXCR7蛋白表達的影響A.單純BMSCs組;B.AMD3100組;C.CXCR7中和抗體組;D.AMD3100+CXCR7中和抗體組,與單純BMSCs組相比,*P<0.05,#P<0.05;與AMD3100組相比,▲P<0.05(x珋±s,n=3)Fig.3WesternblottinganalysisofCXCR4,CXCR7proteinexpressioninBMSCspretreatedwithAMD3100orCXCR7-specificneutralizingantibodyA,BMSCsgroup;B,AMD3100group;C,CXCR7-specificneutralizingantibodygroup;D,AMD3100+CXCR7-specificneutralizingantibodygroup;*P<0.05,#P<0.05,vsBMSCsgroup;▲P<0.05,vsAMD3100group(x珋±s,n=3)
【參考文獻】:
期刊論文
[1]纖維蛋白凝膠與幾丁質對骨髓間充質干細胞向軟骨細胞分化的影響[J]. 李冬梅,金連弘,張寶東,楊水友,李繼承. 解剖學報. 2009 (06)
本文編號:3312265
【文章來源】:解剖學報. 2014,45(05)北大核心CSCD
【文章頁數(shù)】:7 頁
【部分圖文】:
BMSCs的培養(yǎng)與鑒定A.第3代BMSCs;B~D分別為流式細胞術檢測第3代細胞CD44、CD34、CD29表達標尺示100μmFig.1CultureandidentificationofBMSCsA,ThethirdpassageofBMSCs;B-DistheexpressionofCD44,CD34,CD29ofthethirdpassagecellsbyflowcytometryrespectivelyBar=100μm
·642·解剖學報45卷,5期圖1BMSCs的培養(yǎng)與鑒定A.第3代BMSCs;B~D分別為流式細胞術檢測第3代細胞CD44、CD34、CD29表達標尺示100μmFig.1CultureandidentificationofBMSCsA,ThethirdpassageofBMSCs;B-DistheexpressionofCD44,CD34,CD29ofthethirdpassagecellsbyflowcytometryrespectivelyBar=100μm圖2RT-PCR檢測不同處理組細胞CXCR4mRNA、CXCR7mRNA的表達情況M.DNA分子量標準;A.單純BMSCs組;B.AMD3100組;C.CXCR7中和抗體組;D.AMD3100+CXCR7中和抗體組,不同處理組細胞與單純BMSCs組相比,mRNA表達無影響(x珋±s,n=3)Fig.2DetectionofCXCR4mRNA,CXCR7mRNAbyRT-PCRM,DNAmaker;A,BMSCsgroup;B,AMD3100group;C,CXCR7-specificneutralizingantibodygroup;D,AMD3100+CXCR7-specificneutralizingantibodygroup.ComparedwithBMSCsgroup,theexpressionofmRNAarenotchangedbydifferenttreatments(x珋±s,n=3)圖3Westernblotting檢測AMD3100及CXCR7中和抗體對BMSCs中CXCR4、CXCR7蛋白表達的影響A.單純BMSCs組;B.AMD3100組;C.CXCR7中和抗體組;D.AMD3100+CXCR7中和抗體組,與單純BMSCs組相比,*P<0.05,#P<0.05;與AMD3100組相比,▲P<0.05(x珋±s,n=3)Fig.3WesternblottinganalysisofCXCR4,CXCR7proteinexpressioninBMSCspretreatedwithAMD3100orCXCR7-specificneutralizingantibodyA,BMSCsgroup;B,AMD3100group;C,CXCR7-specificneutralizingantibodygroup;D,AMD3100+CXCR7-specificneutralizingantibodygroup;*P<0.05,#P<0.05,vsBMSCsgroup;▲P<0.05,vsAMD3100group(x珋±s,n=3)
·642·解剖學報45卷,5期圖1BMSCs的培養(yǎng)與鑒定A.第3代BMSCs;B~D分別為流式細胞術檢測第3代細胞CD44、CD34、CD29表達標尺示100μmFig.1CultureandidentificationofBMSCsA,ThethirdpassageofBMSCs;B-DistheexpressionofCD44,CD34,CD29ofthethirdpassagecellsbyflowcytometryrespectivelyBar=100μm圖2RT-PCR檢測不同處理組細胞CXCR4mRNA、CXCR7mRNA的表達情況M.DNA分子量標準;A.單純BMSCs組;B.AMD3100組;C.CXCR7中和抗體組;D.AMD3100+CXCR7中和抗體組,不同處理組細胞與單純BMSCs組相比,mRNA表達無影響(x珋±s,n=3)Fig.2DetectionofCXCR4mRNA,CXCR7mRNAbyRT-PCRM,DNAmaker;A,BMSCsgroup;B,AMD3100group;C,CXCR7-specificneutralizingantibodygroup;D,AMD3100+CXCR7-specificneutralizingantibodygroup.ComparedwithBMSCsgroup,theexpressionofmRNAarenotchangedbydifferenttreatments(x珋±s,n=3)圖3Westernblotting檢測AMD3100及CXCR7中和抗體對BMSCs中CXCR4、CXCR7蛋白表達的影響A.單純BMSCs組;B.AMD3100組;C.CXCR7中和抗體組;D.AMD3100+CXCR7中和抗體組,與單純BMSCs組相比,*P<0.05,#P<0.05;與AMD3100組相比,▲P<0.05(x珋±s,n=3)Fig.3WesternblottinganalysisofCXCR4,CXCR7proteinexpressioninBMSCspretreatedwithAMD3100orCXCR7-specificneutralizingantibodyA,BMSCsgroup;B,AMD3100group;C,CXCR7-specificneutralizingantibodygroup;D,AMD3100+CXCR7-specificneutralizingantibodygroup;*P<0.05,#P<0.05,vsBMSCsgroup;▲P<0.05,vsAMD3100group(x珋±s,n=3)
【參考文獻】:
期刊論文
[1]纖維蛋白凝膠與幾丁質對骨髓間充質干細胞向軟骨細胞分化的影響[J]. 李冬梅,金連弘,張寶東,楊水友,李繼承. 解剖學報. 2009 (06)
本文編號:3312265
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