BACKGROUND Heart diseases are the primary cause of death all over the world.Following myocardial infarction,billions of cells die,resulting in a huge loss of cardiac function.Stem cell-based therapies have appeared as...BACKGROUND Heart diseases are the primary cause of death all over the world.Following myocardial infarction,billions of cells die,resulting in a huge loss of cardiac function.Stem cell-based therapies have appeared as a new area to support heart regeneration.The transcription factors GATA binding protein 4(GATA-4)and myocyte enhancer factor 2C(MEF2C)are considered prominent factors in the development of the cardiovascular system.AIM To explore the potential of GATA-4 and MEF2C for the cardiac differentiation of human umbilical cord mesenchymal stem cells(hUC-MSCs).METHODS hUC-MSCs were characterized morphologically and immunologically by the presence of specific markers of MSCs via immunocytochemistry and flow cytometry,and by their potential to differentiate into osteocytes and adipocytes.hUC-MSCs were transfected with GATA-4,MEF2C,and their combination to direct the differentiation.Cardiac differentiation was confirmed by semiquant itative real-time polymerase chain reaction and immunocytochemistry.RESULTS hUC-MSCs expressed specific cell surface markers CD105,CD90,CD44,and vimentin but lack the expression of CD45.The transcription factors GATA-4 and MEF2C,and their combination induced differentiation in hUC-MSCs with significant expression of cardiac genes i.e.,GATA-4,MEF2C,NK2 homeobox 5(NKX2.5),MHC,and connexin-43,and cardiac proteins GATA-4,NKX2.5,cardiac troponin T,and connexin-43.CONCLUSION Transfection with GATA-4,MEF2C,and their combination effectively induces cardiac differentiation in hUC-MSCs.These genetically modified MSCs could be a promising treatment option for heart diseases in the future.展开更多
Objective:To explore the effect of"Yingxiang-Hegu"on Th1,Th2-related cytokines and[2]transcription factors T-bet and GATA-3 in rats with allergic rhinitis.Methods:Rats were randomly divided into three groups...Objective:To explore the effect of"Yingxiang-Hegu"on Th1,Th2-related cytokines and[2]transcription factors T-bet and GATA-3 in rats with allergic rhinitis.Methods:Rats were randomly divided into three groups:blank group,model group and acupoint group.The rat model of ovalbumin(OVA)AR was established,and the general condition of the rats was observed and scored.Acupuncture intervention was performed on the acupoint group on the second day after successful modeling,once per day for 20 min for 10 d.After intervention,the general behavior,behavioral score and histomorphological changes of nasal mucosa were observed.Eosinophils(EOS)were counted under microscope after nasal lavage smear staining,and the contents of total IgE,IFN-γ,IL-12,IL-4 and IL-5 in serum were detected by ELISA.Westernblot and IHC were used to detect the protein level and positive protein expression of specific transcription factors T-bet and GATA-3 in rat nasal mucosa.Results:After the establishment of the model,except for the blank group,the behavioral observation scores of rats in the model group and acupoint group were more than 5 points,indicating that the model was successful.After acupuncture intervention on acupoint"Yingxiang-Hegu",the behavioral score of rats in the acupoint group and western medicine group was significantly lower than that in the model group(P<0.05).Microscopic examination showed that the structure of nasal mucosa in the model group was obviously damaged,cilia were arranged discontinuously,uneven,local congestion and swelling,a large number of epithelial cells exfoliated and necrotic,goblet cell proliferation,obvious inflammatory cell infiltration.The pathological degree of nasal mucosa in the pair point group was significantly less than that in the model group.Compared with the model group,the levels of IFN-γ,IL-2 and IL-12 in serum were significantly increased,while IgE,IL-4,IL-5 and IL-6 were significantly decreased,GATA-3 protein and positive expression in nasal mucosa were significantly decreased and T-bet was significantly increased after acupuncture.Conclusion:Acupuncture at"Yingxiang-Hegu"can effectively improve the nasal sensitive symptoms and control nasal inflammation in AR rats.The mechanism may be that acupuncture at Yingxiang-Hegu can up-regulate the expression of T-bet,decrease the level of GATA-3,promote the production of Th1 cytokines and inhibit the synthesis and secretion of Th2 cells,thus restoring the immune balance of Th1 and Th2.展开更多
目的体外培养HepG2细胞观察川西獐牙菜醇提物(Swertia mussitii Franch)对多药耐药相关蛋白3(multidrug resistance protein 3,MRP3)、核转录因子SP1(specificity protein 1 transcription factor,SP1)及核受体孕烷X受体(pregnane X rec...目的体外培养HepG2细胞观察川西獐牙菜醇提物(Swertia mussitii Franch)对多药耐药相关蛋白3(multidrug resistance protein 3,MRP3)、核转录因子SP1(specificity protein 1 transcription factor,SP1)及核受体孕烷X受体(pregnane X receptor,PXR)、CYP7A启动子结合因子(CYP7A promoter-binding factor,CPF)表达的影响。方法用MTT比色法检测川西獐牙菜醇提物的最佳作用浓度,再分别于0、12、24、48、72h刺激HepG2细胞,抽提细胞的RNA、总蛋白和核蛋白,采用荧光定量PCR和蛋白免疫印迹技术检测膜转运蛋白MRP3、转录因子SP1及核受体PXR、CPF在转录与蛋白水平的表达变化。每个时间点设立阴性对照DMSO组和阳性对照熊去氧胆酸(ursodeoxycholic acid,UDCA)组。结果川西獐牙菜醇提物可显著诱导HepG2细胞膜转运蛋白MRP3的mRNA和蛋白水平的高表达,其作用强于UDCA(P<0.05)。川西獐牙菜醇提物也可明显上调核转录因子SP1及核受体PXR的mRNA和蛋白表达水平,作用强于UDCA。对CPF表达的上调作用与UDCA相当(P<0.05)。结论川西獐牙菜醇提物可刺激HepG2细胞膜转运蛋白MRP3上调,且有可能是通过核转录因子SP1及核受体PXR、CPF上调其表达。展开更多
基金Supported by the Higher Education Commission(HEC),Pakistan Scholarship for Ph.D.Studies to Razzaq SS,No.520-148390-2BS6-011.
文摘BACKGROUND Heart diseases are the primary cause of death all over the world.Following myocardial infarction,billions of cells die,resulting in a huge loss of cardiac function.Stem cell-based therapies have appeared as a new area to support heart regeneration.The transcription factors GATA binding protein 4(GATA-4)and myocyte enhancer factor 2C(MEF2C)are considered prominent factors in the development of the cardiovascular system.AIM To explore the potential of GATA-4 and MEF2C for the cardiac differentiation of human umbilical cord mesenchymal stem cells(hUC-MSCs).METHODS hUC-MSCs were characterized morphologically and immunologically by the presence of specific markers of MSCs via immunocytochemistry and flow cytometry,and by their potential to differentiate into osteocytes and adipocytes.hUC-MSCs were transfected with GATA-4,MEF2C,and their combination to direct the differentiation.Cardiac differentiation was confirmed by semiquant itative real-time polymerase chain reaction and immunocytochemistry.RESULTS hUC-MSCs expressed specific cell surface markers CD105,CD90,CD44,and vimentin but lack the expression of CD45.The transcription factors GATA-4 and MEF2C,and their combination induced differentiation in hUC-MSCs with significant expression of cardiac genes i.e.,GATA-4,MEF2C,NK2 homeobox 5(NKX2.5),MHC,and connexin-43,and cardiac proteins GATA-4,NKX2.5,cardiac troponin T,and connexin-43.CONCLUSION Transfection with GATA-4,MEF2C,and their combination effectively induces cardiac differentiation in hUC-MSCs.These genetically modified MSCs could be a promising treatment option for heart diseases in the future.
基金Lv Jingshan Acupuncture and Medicine Combination Technology Innovation Team(No.2019TD-003)Lv Jingshan Acupuncture and Moxibustion and Bioelectronic Medicine Innovation Team(No.2022TD-1006)。
文摘Objective:To explore the effect of"Yingxiang-Hegu"on Th1,Th2-related cytokines and[2]transcription factors T-bet and GATA-3 in rats with allergic rhinitis.Methods:Rats were randomly divided into three groups:blank group,model group and acupoint group.The rat model of ovalbumin(OVA)AR was established,and the general condition of the rats was observed and scored.Acupuncture intervention was performed on the acupoint group on the second day after successful modeling,once per day for 20 min for 10 d.After intervention,the general behavior,behavioral score and histomorphological changes of nasal mucosa were observed.Eosinophils(EOS)were counted under microscope after nasal lavage smear staining,and the contents of total IgE,IFN-γ,IL-12,IL-4 and IL-5 in serum were detected by ELISA.Westernblot and IHC were used to detect the protein level and positive protein expression of specific transcription factors T-bet and GATA-3 in rat nasal mucosa.Results:After the establishment of the model,except for the blank group,the behavioral observation scores of rats in the model group and acupoint group were more than 5 points,indicating that the model was successful.After acupuncture intervention on acupoint"Yingxiang-Hegu",the behavioral score of rats in the acupoint group and western medicine group was significantly lower than that in the model group(P<0.05).Microscopic examination showed that the structure of nasal mucosa in the model group was obviously damaged,cilia were arranged discontinuously,uneven,local congestion and swelling,a large number of epithelial cells exfoliated and necrotic,goblet cell proliferation,obvious inflammatory cell infiltration.The pathological degree of nasal mucosa in the pair point group was significantly less than that in the model group.Compared with the model group,the levels of IFN-γ,IL-2 and IL-12 in serum were significantly increased,while IgE,IL-4,IL-5 and IL-6 were significantly decreased,GATA-3 protein and positive expression in nasal mucosa were significantly decreased and T-bet was significantly increased after acupuncture.Conclusion:Acupuncture at"Yingxiang-Hegu"can effectively improve the nasal sensitive symptoms and control nasal inflammation in AR rats.The mechanism may be that acupuncture at Yingxiang-Hegu can up-regulate the expression of T-bet,decrease the level of GATA-3,promote the production of Th1 cytokines and inhibit the synthesis and secretion of Th2 cells,thus restoring the immune balance of Th1 and Th2.
文摘目的体外培养HepG2细胞观察川西獐牙菜醇提物(Swertia mussitii Franch)对多药耐药相关蛋白3(multidrug resistance protein 3,MRP3)、核转录因子SP1(specificity protein 1 transcription factor,SP1)及核受体孕烷X受体(pregnane X receptor,PXR)、CYP7A启动子结合因子(CYP7A promoter-binding factor,CPF)表达的影响。方法用MTT比色法检测川西獐牙菜醇提物的最佳作用浓度,再分别于0、12、24、48、72h刺激HepG2细胞,抽提细胞的RNA、总蛋白和核蛋白,采用荧光定量PCR和蛋白免疫印迹技术检测膜转运蛋白MRP3、转录因子SP1及核受体PXR、CPF在转录与蛋白水平的表达变化。每个时间点设立阴性对照DMSO组和阳性对照熊去氧胆酸(ursodeoxycholic acid,UDCA)组。结果川西獐牙菜醇提物可显著诱导HepG2细胞膜转运蛋白MRP3的mRNA和蛋白水平的高表达,其作用强于UDCA(P<0.05)。川西獐牙菜醇提物也可明显上调核转录因子SP1及核受体PXR的mRNA和蛋白表达水平,作用强于UDCA。对CPF表达的上调作用与UDCA相当(P<0.05)。结论川西獐牙菜醇提物可刺激HepG2细胞膜转运蛋白MRP3上调,且有可能是通过核转录因子SP1及核受体PXR、CPF上调其表达。
