Objective:To investigate the effect of SHU555A,a clinically approved iron nanoparticle,labeling on differentiation of bone marrow mesenchymal stem cells(BMSCs) into neurocyte-like cells in vitro.Methods:10 times dilut...Objective:To investigate the effect of SHU555A,a clinically approved iron nanoparticle,labeling on differentiation of bone marrow mesenchymal stem cells(BMSCs) into neurocyte-like cells in vitro.Methods:10 times dilution of 10μl,20μl,40μl and 80μl SHU555A were added to 2ml of culture medium containing rat BMSCs to obtain four experimental groups of SHU555A labeling of BMSCs with ferri ion concentrations of 14μg/ml,28μg/ml,56μg/ml and 112μg/ml,respectively.2ml of culture medium with rat BMSCs did not contain SHU555A served as control group.The BMSCs of all the groups were pre-induced by bFGF,and induced by DMSO/butylated hydroxyanisole(BHA) for six hours,subsequently reverse transcription polymerase chain reaction(RT-PCR) technique was employed to detect mRNA expression of nestin,neuronspecific analase(NSE) and glial fibrillary acid protein(GFAP).Western blot technique was used to detectprotein expression of nestin.Results:Quantitative-PCR revealed high mRNA expression of nestin,NSE and GFAP induced by DMSO/BHA in all the experimental groups,but the difference between the experimental groups and the control group was not significant(P>0.05).Western blot analysis demonstrated there was no statistically significant difference in nestin protein expression between the experimental groups and the control group(P>0.05).Conclusion:SHU555A labeling do not affect differentiation of rat BMSCs into neurocyte-like cells in vitro.展开更多
Aim Aging is an independent risk factor for heart disease, however the effective intervention has not been found so far. Bone marrow mesenchymal stem cells （BMSCs） have been shown to offer a wide variety of cel- l...Aim Aging is an independent risk factor for heart disease, however the effective intervention has not been found so far. Bone marrow mesenchymal stem cells （BMSCs） have been shown to offer a wide variety of cel- lular functions including the protective effects on damaged hearts. Here we investigated the antiaging properties of BMSCs and the underlying mechanism in a cellular model of cardiomyocyte senescence and a rat model of aging hearts. Methods In vitro study, neonatal rat ventricular cells （NRVCs） and BMSCs were cocultured in the same dish with a semipermeable membrane to separate the two populations. In vivo, the BMSCs were injected into the rat hearts to observe their antiaging effects. The expression of β-galactosidase and aging-related proteins, and the lev- els of oxidative stress were determined in vivo and in vitro. The heart function was measured by the High-Resolution Imaging System. Results Monocultured NRVCs displayed the senescence-associated phenotypes, characterized by an increase in the number of β-galaetosidase-positive cells and decreases in the degradation and disappearance of cellular organelles in a time-dependent manner. The levels of reactive oxygen species and malondialdehyde were el- evated, whereas the activities of antioxidant enzymes superoxide dismutase and glutathione peroxidase were de- creased, along with upregulation of p53, p21cipl/wafl and p16INK4a in the aging eardiomyoeytes. These deleterious alterations were abrogated in aging NRVCs cocultured with BMSCs. Qualitatively, the same senescent phenotypes were consistently observed in aging rat hearts. Notably, BMSC transplantation significantly prevented these detri- mental alterations and improved the impaired cardiac function in the aging rats. Conclusions BMSCs possess strong antiseneseence action on the aging NRVCs and hearts and can improve cardiac function after transplantation in aging rats. The present study, therefore, provides an alternative approach for the treatment of heart failure in the elderly population.展开更多
目的观察辛伐他汀(SIM)体外单独给药对大鼠骨髓间充质干细胞(BMSCs)成骨分化的影响。方法取第二代大鼠BMSCs随机分为4组,对照组(CM):完全培养基培养;诱导组(OM):成骨诱导培养基培养;辛伐他汀组(SIM):含终浓度为10-7mol/L的辛伐他汀的完...目的观察辛伐他汀(SIM)体外单独给药对大鼠骨髓间充质干细胞(BMSCs)成骨分化的影响。方法取第二代大鼠BMSCs随机分为4组,对照组(CM):完全培养基培养;诱导组(OM):成骨诱导培养基培养;辛伐他汀组(SIM):含终浓度为10-7mol/L的辛伐他汀的完全培养基培养;阻断剂组(SB+SIM):先用p38MAPK通路阻断剂SB203580干预30min后,加入等浓度辛伐他汀。给药4 h后,Western Blot检测p-p38MAPK、p38MAPK蛋白的表达。给药7d后,进行碱性磷酸酶(ALP)染色和比活性测定。给药7、14d后,采用Real-time PCR法检测ALP、Ι型胶原(COLΙ)、骨钙素(OCN)和Runt相关转录因子2(Runx2)m RNA的表达。结果 1Western Blot检测:OM组和SIM组p-p38MAPK表达水平显著高于CM组和SB+SIM组(P<0.05)。2ALP比活性测定:OM组和SIM组显著高于CM组,与相比,SB+SIM组显著低于SIM组(P<0.05)。3ALP染色:OM组和SIM组细胞胞浆内出现棕色颗粒,而SB+SIM组和CM组未见明显阳性染色颗粒。4Real-time PCR检测:在第7d、14d,OM组和SIM组ALP、COLΙ和Runx2 m RNA表达水平均显著高于CM组,SB+SIM组明显低于SIM组(P<0.05);第7d,OM组OCN m RNA表达水平明显高于CM组(P<0.05);在第14d,OM组和SIM组OCN m RNA表达水平高于CM组,SB+SIM组低于SIM组(P<0.05)。结论辛伐他汀可以在缺乏成骨诱导成分的环境下,通过活化p38MAPK信号诱导大鼠BMSCs向成骨细胞分化。展开更多
基金Henan ontstanding talent program(084200510012)zhou research programs(083SGYS33262-5)zhou university 2011 project,third constraction projection:basic and clinical research of stem cell
文摘Objective:To investigate the effect of SHU555A,a clinically approved iron nanoparticle,labeling on differentiation of bone marrow mesenchymal stem cells(BMSCs) into neurocyte-like cells in vitro.Methods:10 times dilution of 10μl,20μl,40μl and 80μl SHU555A were added to 2ml of culture medium containing rat BMSCs to obtain four experimental groups of SHU555A labeling of BMSCs with ferri ion concentrations of 14μg/ml,28μg/ml,56μg/ml and 112μg/ml,respectively.2ml of culture medium with rat BMSCs did not contain SHU555A served as control group.The BMSCs of all the groups were pre-induced by bFGF,and induced by DMSO/butylated hydroxyanisole(BHA) for six hours,subsequently reverse transcription polymerase chain reaction(RT-PCR) technique was employed to detect mRNA expression of nestin,neuronspecific analase(NSE) and glial fibrillary acid protein(GFAP).Western blot technique was used to detectprotein expression of nestin.Results:Quantitative-PCR revealed high mRNA expression of nestin,NSE and GFAP induced by DMSO/BHA in all the experimental groups,but the difference between the experimental groups and the control group was not significant(P>0.05).Western blot analysis demonstrated there was no statistically significant difference in nestin protein expression between the experimental groups and the control group(P>0.05).Conclusion:SHU555A labeling do not affect differentiation of rat BMSCs into neurocyte-like cells in vitro.
文摘Aim Aging is an independent risk factor for heart disease, however the effective intervention has not been found so far. Bone marrow mesenchymal stem cells （BMSCs） have been shown to offer a wide variety of cel- lular functions including the protective effects on damaged hearts. Here we investigated the antiaging properties of BMSCs and the underlying mechanism in a cellular model of cardiomyocyte senescence and a rat model of aging hearts. Methods In vitro study, neonatal rat ventricular cells （NRVCs） and BMSCs were cocultured in the same dish with a semipermeable membrane to separate the two populations. In vivo, the BMSCs were injected into the rat hearts to observe their antiaging effects. The expression of β-galactosidase and aging-related proteins, and the lev- els of oxidative stress were determined in vivo and in vitro. The heart function was measured by the High-Resolution Imaging System. Results Monocultured NRVCs displayed the senescence-associated phenotypes, characterized by an increase in the number of β-galaetosidase-positive cells and decreases in the degradation and disappearance of cellular organelles in a time-dependent manner. The levels of reactive oxygen species and malondialdehyde were el- evated, whereas the activities of antioxidant enzymes superoxide dismutase and glutathione peroxidase were de- creased, along with upregulation of p53, p21cipl/wafl and p16INK4a in the aging eardiomyoeytes. These deleterious alterations were abrogated in aging NRVCs cocultured with BMSCs. Qualitatively, the same senescent phenotypes were consistently observed in aging rat hearts. Notably, BMSC transplantation significantly prevented these detri- mental alterations and improved the impaired cardiac function in the aging rats. Conclusions BMSCs possess strong antiseneseence action on the aging NRVCs and hearts and can improve cardiac function after transplantation in aging rats. The present study, therefore, provides an alternative approach for the treatment of heart failure in the elderly population.
文摘目的观察辛伐他汀(SIM)体外单独给药对大鼠骨髓间充质干细胞(BMSCs)成骨分化的影响。方法取第二代大鼠BMSCs随机分为4组,对照组(CM):完全培养基培养;诱导组(OM):成骨诱导培养基培养;辛伐他汀组(SIM):含终浓度为10-7mol/L的辛伐他汀的完全培养基培养;阻断剂组(SB+SIM):先用p38MAPK通路阻断剂SB203580干预30min后,加入等浓度辛伐他汀。给药4 h后,Western Blot检测p-p38MAPK、p38MAPK蛋白的表达。给药7d后,进行碱性磷酸酶(ALP)染色和比活性测定。给药7、14d后,采用Real-time PCR法检测ALP、Ι型胶原(COLΙ)、骨钙素(OCN)和Runt相关转录因子2(Runx2)m RNA的表达。结果 1Western Blot检测:OM组和SIM组p-p38MAPK表达水平显著高于CM组和SB+SIM组(P<0.05)。2ALP比活性测定:OM组和SIM组显著高于CM组,与相比,SB+SIM组显著低于SIM组(P<0.05)。3ALP染色:OM组和SIM组细胞胞浆内出现棕色颗粒,而SB+SIM组和CM组未见明显阳性染色颗粒。4Real-time PCR检测:在第7d、14d,OM组和SIM组ALP、COLΙ和Runx2 m RNA表达水平均显著高于CM组,SB+SIM组明显低于SIM组(P<0.05);第7d,OM组OCN m RNA表达水平明显高于CM组(P<0.05);在第14d,OM组和SIM组OCN m RNA表达水平高于CM组,SB+SIM组低于SIM组(P<0.05)。结论辛伐他汀可以在缺乏成骨诱导成分的环境下,通过活化p38MAPK信号诱导大鼠BMSCs向成骨细胞分化。
