为探讨去卵巢骨质疏松大鼠骨髓间充质干细胞(Bone m arrow m esenchym a l stem ce lls,M SC s)成纤维细胞集落生成单位(CFU-F s)、细胞周期、成骨分化能力和成脂分化能力的变化,选用3月龄和6月龄健康雌性SD大鼠行双侧卵巢切除术,建立...为探讨去卵巢骨质疏松大鼠骨髓间充质干细胞(Bone m arrow m esenchym a l stem ce lls,M SC s)成纤维细胞集落生成单位(CFU-F s)、细胞周期、成骨分化能力和成脂分化能力的变化,选用3月龄和6月龄健康雌性SD大鼠行双侧卵巢切除术,建立骨质疏松动物模型,动物实验分为4组:(1)3月龄正常组(con tro l-3);(2)3月龄卵巢切除组(OVX-3);(3)6月龄正常组(con tro l-6);(4)6月龄卵巢切除组(OVX-6)。采用密度梯度离心方法分别获取各组M SC s,体外培养,选用第3~4代M SC s用于实验。倒置相差显微镜观察CFU-F s;流式细胞技术检测细胞周期、细胞增殖指数(pro liferation index,P I)和细胞凋亡指数(apoptotic index,A I))。加入成骨诱导剂,采用碱性磷酸酶活性蛋白检测法(IFCC推荐法)检测M SC s碱性磷酸酶(a lka line phosphatase,ALP)分泌量;采用同位素标记方法检测骨钙素(O steoca lc in,OCN)分泌量;茜素红染色观察钙结节形成。加入成脂诱导剂,油红O染色观察M SC s内脂滴形成;RT-PCR方法检测M SC s脂蛋白脂酶(lipoprote in lipase,LPL)mRNA的表达。结果表明:与相应对照组比较,OVX-3组和OVX-6组CFU-F s、P I值明显降低(P<0.05),A I值明显增加(P<0.05);OVX-6组CFU-F s、P I值的降低和A I值的增加都明显大于OVX-3组(P<0.05)。成骨诱导表明:与相应对照组比较,OVX-3组和OVX-6组ALP、BGP分泌量以及钙结节形成的数量均明显降低(P<0.05),OVX-6组的降低最为明显。成脂诱导表明:与相应对照组比较,OVX-3组和OVX-6组,LPL mRNA的表达量明显增加(P<0.05),OVX-6组LPL mRNA的表达量的增加明显高于其他各组(P<0.05)。结论:去卵巢骨质疏松大鼠M SC s增殖能力明显下降,向成骨细胞分化能力下降,向脂肪细胞分化能力增强,其中6OVX组M SC s的变化最明显。提示:去卵巢骨质疏松大鼠M SC s向成骨细胞分化能力的降低和向脂肪细胞分化能力的增强,可能与绝经后骨质疏松症的发生相关。展开更多
目的观察缺氧预处理骨髓间充质干细胞(MSCs)移植对脑缺血再灌注损伤大鼠SDF-1/CXCR4 m RNA和蛋白表达的影响及清热化瘀方的干预作用。方法采用线栓法制备局灶性脑缺血大鼠(MCAO)模型,将216只SD大鼠随机分为6组:假手术组(SO)、模型组(MC...目的观察缺氧预处理骨髓间充质干细胞(MSCs)移植对脑缺血再灌注损伤大鼠SDF-1/CXCR4 m RNA和蛋白表达的影响及清热化瘀方的干预作用。方法采用线栓法制备局灶性脑缺血大鼠(MCAO)模型,将216只SD大鼠随机分为6组:假手术组(SO)、模型组(MCAO)、MSCs移植对照组(N-MSCs)、经HP处理后的MSCs移植组(HP-MSCs)、MSCs移植联合清热化瘀方组(MSCs+QRHY)、HP-MSCs移植联合清热化瘀方组(HP+QRHY)。每组大鼠36只,每组根据取材时间点7,14,28 d又可分为每组3个亚组,每个亚组12只大鼠。采用q RT-PCR和Western blot观察3个时间点SDF-1/CXCR4 m RNA及其蛋白的表达变化,并以TUNEL法检测神经细胞凋亡。结果各组缺血半暗带SDF-1/CXCR4 m RNA及其蛋白的表达均于7d达到高峰,14,28 d表达逐渐下降。其中7,14 d同一时间点组间比较,HP-MSCs组、MSCs+QRHY组及HP+QRHY组二者的表达均明显优于N-MSCs组(P<0.01,P<0.05),而以HP+QRHY组增高最为明显(P<0.05,P<0.01),28 d后,各移植组的表达趋势趋同,但仍高于模型组(P<0.05)。结论缺氧预处理MSCs移植能够显著提高脑缺血再灌注损伤大鼠SDF-1/CXCR4的表达,清热化瘀方能够进一步上调SDF/1CXCR4的表达,减少细胞凋亡。展开更多
Mesenchymal stem cells （MSCs） of nonembryonic origins possess the proliferation and multi-lineage differentiation potentials. It has been established that epigenetic mechanisms could be critical for determining the ...Mesenchymal stem cells （MSCs） of nonembryonic origins possess the proliferation and multi-lineage differentiation potentials. It has been established that epigenetic mechanisms could be critical for determining the fate of stem cells, and MSCs derived from different origins exhibited different expression profiles individually to a certain extent. In this study, ChiP-on-chip was used to generate genome-wide histone H3-Lys9 acetylation and dimethylation profiles at gene promoters in human bone marrow MSCs. We showed that modifications of histone H3-Lys9 at gene promoters correlated well with mRNA expression in human bone marrow MSCs. Functional analysis revealed that many key cellular pathways in human bone marrow MSC self-renewal, such as the canonical signaling pathways, cell cycle pathways and cytokine related pathways may be regulated by H3-Lys9 modifications. These data suggest that gene activation and silencing affected by H3-Lys9 acetylation and dimethylation, respectively, may be essential to the maintenance of human bone marrow MSC self-renewal and multi-potency.展开更多
文摘目的观察缺氧预处理骨髓间充质干细胞(MSCs)移植对脑缺血再灌注损伤大鼠SDF-1/CXCR4 m RNA和蛋白表达的影响及清热化瘀方的干预作用。方法采用线栓法制备局灶性脑缺血大鼠(MCAO)模型,将216只SD大鼠随机分为6组:假手术组(SO)、模型组(MCAO)、MSCs移植对照组(N-MSCs)、经HP处理后的MSCs移植组(HP-MSCs)、MSCs移植联合清热化瘀方组(MSCs+QRHY)、HP-MSCs移植联合清热化瘀方组(HP+QRHY)。每组大鼠36只,每组根据取材时间点7,14,28 d又可分为每组3个亚组,每个亚组12只大鼠。采用q RT-PCR和Western blot观察3个时间点SDF-1/CXCR4 m RNA及其蛋白的表达变化,并以TUNEL法检测神经细胞凋亡。结果各组缺血半暗带SDF-1/CXCR4 m RNA及其蛋白的表达均于7d达到高峰,14,28 d表达逐渐下降。其中7,14 d同一时间点组间比较,HP-MSCs组、MSCs+QRHY组及HP+QRHY组二者的表达均明显优于N-MSCs组(P<0.01,P<0.05),而以HP+QRHY组增高最为明显(P<0.05,P<0.01),28 d后,各移植组的表达趋势趋同,但仍高于模型组(P<0.05)。结论缺氧预处理MSCs移植能够显著提高脑缺血再灌注损伤大鼠SDF-1/CXCR4的表达,清热化瘀方能够进一步上调SDF/1CXCR4的表达,减少细胞凋亡。
基金the National Basic Research Program of China (No 2005CB522404 and 2006CB910506)the Program for Changjiang Scholars and Innovative Research Team (PCSIRT) in Universities (No IRT0519)the National Natural Science Founda-tion of China (No 30771232 and 30671184)
文摘Mesenchymal stem cells （MSCs） of nonembryonic origins possess the proliferation and multi-lineage differentiation potentials. It has been established that epigenetic mechanisms could be critical for determining the fate of stem cells, and MSCs derived from different origins exhibited different expression profiles individually to a certain extent. In this study, ChiP-on-chip was used to generate genome-wide histone H3-Lys9 acetylation and dimethylation profiles at gene promoters in human bone marrow MSCs. We showed that modifications of histone H3-Lys9 at gene promoters correlated well with mRNA expression in human bone marrow MSCs. Functional analysis revealed that many key cellular pathways in human bone marrow MSC self-renewal, such as the canonical signaling pathways, cell cycle pathways and cytokine related pathways may be regulated by H3-Lys9 modifications. These data suggest that gene activation and silencing affected by H3-Lys9 acetylation and dimethylation, respectively, may be essential to the maintenance of human bone marrow MSC self-renewal and multi-potency.
