目的：研究柴胡三参胶囊含药血清对骨髓间充质干细胞分化为心肌细胞过程中PI3-K/Akt信号通路的影响,探讨其抗心律失常机制与PI3-K/Akt信号转导通路的相关性。方法：将P3代骨髓间充质干细胞随机分为4组,分别为BMP-2组、BMP-2＋含药血清...目的：研究柴胡三参胶囊含药血清对骨髓间充质干细胞分化为心肌细胞过程中PI3-K/Akt信号通路的影响,探讨其抗心律失常机制与PI3-K/Akt信号转导通路的相关性。方法：将P3代骨髓间充质干细胞随机分为4组,分别为BMP-2组、BMP-2＋含药血清组、含药血清组及空白血清组,以MTT法测BMSCs增殖情况,确定最佳浓度及最佳时间;加入相应浓度的诱导剂及血清,诱导至最佳时间,Western blotting法检测各组细胞内Akt蛋白表达水平;RT-PCR法检测各组细胞内GATA-4、β-MHC m RNA表达情况。结果：与空白血清组相比,BMP-2组、BMP-2＋含药血清组、含药血清组Akt、GATA-4、β-MHC表达水平均明显升高,统计学具有明显差异（P〈0.01）;与BMP-2组相比,BMP-2＋含药血清组表达水平升高（P〈0.05）,与含药血清组相比,BMP-2＋含药血清组表达水平升高（P〈0.05）;BMP-2组与含药血清组比较,差异无统计学意义（P〉0.05）。结论：柴胡三参胶囊能够通过作用于PI3-K/Akt信号通路诱导骨髓间充质干细胞向心肌细胞分化。展开更多
Glycogen synthase kinase-3β (GSK-3β) is a serine/threonine protein kinase,which takes part not only in glycogen metabolism,but also in cell proliferation,differentiation and apoptosis. GSK-3β is inhibited by growth...Glycogen synthase kinase-3β (GSK-3β) is a serine/threonine protein kinase,which takes part not only in glycogen metabolism,but also in cell proliferation,differentiation and apoptosis. GSK-3β is inhibited by growth factors and hypertrophic stimuli through phosphorylation of its N-terminal end serine (Ser9) residue. It is also activated by phosphorylation of its tyrosine (Tyr216) residue. GSK-3β is profoundly inactivated in mice with hypertrophic cardiomyopathy (HCM) and plays a secondary role in myosin heavy chain mutation. However,the role of GSK-3β in HCM was controversial. Recent studies have demonstrated that the activation of GSK-3β inhibits the myocardial hypertrophy,and is regulated by Wnt/Frizzld and PI3-K/Akt signaling pathways. This article introduces the molecular role of glycogen synthase kinase-3β signaling in myocardial hypertrophy and the different pathways on the activity of glycogen synthase kinase-3β (GSK-3β)展开更多
目的了解六黄合剂对胰岛素抵抗大鼠血管内皮保护的作用机制。方法健康SD大鼠,按体重随机分为空白组、模型组、六黄合剂组和罗格列酮组,每组10只。应用高脂饲料联合肌注地塞米松的方法诱导大鼠胰岛素抵抗模型,分别给予罗格列酮和六黄合...目的了解六黄合剂对胰岛素抵抗大鼠血管内皮保护的作用机制。方法健康SD大鼠,按体重随机分为空白组、模型组、六黄合剂组和罗格列酮组,每组10只。应用高脂饲料联合肌注地塞米松的方法诱导大鼠胰岛素抵抗模型,分别给予罗格列酮和六黄合剂干预,7周后应用RT-PCR法测定大鼠胸主动脉PI3-Km RNA、Aktm RNA和e NOS m RNA的表达水平。结果与模型组比较,六黄合剂组胸主动脉Aktm RNA表达水平(0.20±0.05)明显升高(P<0.05),PI3-Km RNA和e NOS m RNA的表达水平也升高,分别为(1.07±0.31)和(0.87±0.27),差异无统计学意义。结论六黄合剂通过上调PI3-K/Akt信号通路来发挥对胰岛素抵抗大鼠血管内皮的保护作用。展开更多
Objective The present study aimed to explore the role of P2Y1 receptor in glial fibrillary acidic protein （GFAP） production and glial cell line-derived neurotrophic factor （GDNF） secretion of astrocytes under isch...Objective The present study aimed to explore the role of P2Y1 receptor in glial fibrillary acidic protein （GFAP） production and glial cell line-derived neurotrophic factor （GDNF） secretion of astrocytes under ischemic insult and the related signaling pathways. Methods Using transient right middle cerebral artery occlusion （tMCAO） and oxygen-glucose-serum deprivation for 2 h as the model of ischemic injury in vivo and in vitro, immunofluorescence, quantitative real-time reverse transcription-polymerase chain reaction （RT-PCR）, Western blotting, enzyme linked immunosorbent assay （ELISA） were used to investigate location of P2Y1 receptor and GDNF, the expression of GFAP and GDNF, and the changes of signaling molecules. Results Blockage of P2Y1 receptor with the selective antagonist N^6-methyl-2′-deoxyadenosine 3′,5′-bisphosphate diammonium （MRS2179） reduced GFAP production and increased GDNF production in the antagonist group as compared with simple ischemic group both in vivo and in vitro. Oxygen-glucose-serum deprivation and blockage of P2Y1 receptor caused elevation of phosphorylated Akt and cAMP response element binding protein （CREB）, and reduction of phosphorylated Janus kinase2 （JAK2） and signal transducer and activator of transcription3 （STAT3, Ser727）. After blockage of P2Y1 receptor and deprivation of oxygen-glucose-serum, AG490 （inhibitor of JAK2） reduced phosphorylation of STAT3 （Ser727） as well as expression of GFAP; LY294002, an inhibitor of phosphatidylinositol 3-kinase （PI3-K）, decreased phosphorylation of Akt and CREB; the inhibitor of mitogen-activated protein kinase kinase 1/2 （MEK 1/2） U0126, an important molecule of Ras/extracellular signal- regulated kinase （ERK） signaling pathway, decreased the phosphorylation of JAK2, STAT3 （Ser727）, Akt and CREB. Conclusion These results suggest that P2Y1 receptor plays a role in the production of GFAP and GDNF in astrocytes under transient ischemic condition and the related signaling pathways may be JAK2/STAT3 and PI3-K/Akt/CREB, respectively, and that crosstalk probably exists between them.展开更多
文摘目的：研究柴胡三参胶囊含药血清对骨髓间充质干细胞分化为心肌细胞过程中PI3-K/Akt信号通路的影响,探讨其抗心律失常机制与PI3-K/Akt信号转导通路的相关性。方法：将P3代骨髓间充质干细胞随机分为4组,分别为BMP-2组、BMP-2＋含药血清组、含药血清组及空白血清组,以MTT法测BMSCs增殖情况,确定最佳浓度及最佳时间;加入相应浓度的诱导剂及血清,诱导至最佳时间,Western blotting法检测各组细胞内Akt蛋白表达水平;RT-PCR法检测各组细胞内GATA-4、β-MHC m RNA表达情况。结果：与空白血清组相比,BMP-2组、BMP-2＋含药血清组、含药血清组Akt、GATA-4、β-MHC表达水平均明显升高,统计学具有明显差异（P〈0.01）;与BMP-2组相比,BMP-2＋含药血清组表达水平升高（P〈0.05）,与含药血清组相比,BMP-2＋含药血清组表达水平升高（P〈0.05）;BMP-2组与含药血清组比较,差异无统计学意义（P〉0.05）。结论：柴胡三参胶囊能够通过作用于PI3-K/Akt信号通路诱导骨髓间充质干细胞向心肌细胞分化。
文摘Glycogen synthase kinase-3β (GSK-3β) is a serine/threonine protein kinase,which takes part not only in glycogen metabolism,but also in cell proliferation,differentiation and apoptosis. GSK-3β is inhibited by growth factors and hypertrophic stimuli through phosphorylation of its N-terminal end serine (Ser9) residue. It is also activated by phosphorylation of its tyrosine (Tyr216) residue. GSK-3β is profoundly inactivated in mice with hypertrophic cardiomyopathy (HCM) and plays a secondary role in myosin heavy chain mutation. However,the role of GSK-3β in HCM was controversial. Recent studies have demonstrated that the activation of GSK-3β inhibits the myocardial hypertrophy,and is regulated by Wnt/Frizzld and PI3-K/Akt signaling pathways. This article introduces the molecular role of glycogen synthase kinase-3β signaling in myocardial hypertrophy and the different pathways on the activity of glycogen synthase kinase-3β (GSK-3β)
文摘目的了解六黄合剂对胰岛素抵抗大鼠血管内皮保护的作用机制。方法健康SD大鼠,按体重随机分为空白组、模型组、六黄合剂组和罗格列酮组,每组10只。应用高脂饲料联合肌注地塞米松的方法诱导大鼠胰岛素抵抗模型,分别给予罗格列酮和六黄合剂干预,7周后应用RT-PCR法测定大鼠胸主动脉PI3-Km RNA、Aktm RNA和e NOS m RNA的表达水平。结果与模型组比较,六黄合剂组胸主动脉Aktm RNA表达水平(0.20±0.05)明显升高(P<0.05),PI3-Km RNA和e NOS m RNA的表达水平也升高,分别为(1.07±0.31)和(0.87±0.27),差异无统计学意义。结论六黄合剂通过上调PI3-K/Akt信号通路来发挥对胰岛素抵抗大鼠血管内皮的保护作用。
基金the National Natural Science Foundation of China (No. 30500189)
文摘Objective The present study aimed to explore the role of P2Y1 receptor in glial fibrillary acidic protein （GFAP） production and glial cell line-derived neurotrophic factor （GDNF） secretion of astrocytes under ischemic insult and the related signaling pathways. Methods Using transient right middle cerebral artery occlusion （tMCAO） and oxygen-glucose-serum deprivation for 2 h as the model of ischemic injury in vivo and in vitro, immunofluorescence, quantitative real-time reverse transcription-polymerase chain reaction （RT-PCR）, Western blotting, enzyme linked immunosorbent assay （ELISA） were used to investigate location of P2Y1 receptor and GDNF, the expression of GFAP and GDNF, and the changes of signaling molecules. Results Blockage of P2Y1 receptor with the selective antagonist N^6-methyl-2′-deoxyadenosine 3′,5′-bisphosphate diammonium （MRS2179） reduced GFAP production and increased GDNF production in the antagonist group as compared with simple ischemic group both in vivo and in vitro. Oxygen-glucose-serum deprivation and blockage of P2Y1 receptor caused elevation of phosphorylated Akt and cAMP response element binding protein （CREB）, and reduction of phosphorylated Janus kinase2 （JAK2） and signal transducer and activator of transcription3 （STAT3, Ser727）. After blockage of P2Y1 receptor and deprivation of oxygen-glucose-serum, AG490 （inhibitor of JAK2） reduced phosphorylation of STAT3 （Ser727） as well as expression of GFAP; LY294002, an inhibitor of phosphatidylinositol 3-kinase （PI3-K）, decreased phosphorylation of Akt and CREB; the inhibitor of mitogen-activated protein kinase kinase 1/2 （MEK 1/2） U0126, an important molecule of Ras/extracellular signal- regulated kinase （ERK） signaling pathway, decreased the phosphorylation of JAK2, STAT3 （Ser727）, Akt and CREB. Conclusion These results suggest that P2Y1 receptor plays a role in the production of GFAP and GDNF in astrocytes under transient ischemic condition and the related signaling pathways may be JAK2/STAT3 and PI3-K/Akt/CREB, respectively, and that crosstalk probably exists between them.
