Objective:To investigate the mechanism by which hydroxyl safflower yellow A,an active component of safflower (Carthamus tinctorius L.),promotes apoptosis in abnormal human umbilical vein endothelial cells (HUVECs).Met...Objective:To investigate the mechanism by which hydroxyl safflower yellow A,an active component of safflower (Carthamus tinctorius L.),promotes apoptosis in abnormal human umbilical vein endothelial cells (HUVECs).Methods:Supernatant of BGC-823 was used to stimulate HUVECs to establish a model of abnormal proliferation of HUVECs.After determining an ideal concentration of HSYA by MTT assay,apoptosis was detected with flow cytometry and TUNEL assay.Mechanism of apoptosis was assessed using quantitative real-time polymerase chain reaction,Western blot,and ELISA.Results:A range of concentrations of HSYA inhibited proliferation and promoted apoptosis of abnormal HUVECs.As the rate of apoptosis increased,mRNA expression of caspase-3 increased while expression of mutant p53 decreased.HSYA had no effect on Fas gene expression.Analogously,protein expression of Bax was increased while those of Bcl-2,Fas,and Fas-L were decreased.Conclusions:HSYA appears to induce apoptosis of HUVECs with the stimulation of the supematant of tumor cells.The mechanism of apoptosis by HSYA may involve activation of the mitochondrial apoptotic pathway and regulation of the expressions of Bcl-2,Bax,and p53.展开更多
Objective To study the protective effects of hydroxysafflor yellow A （HSYA） against the oxidative damage caused by β-mercaptoethanol （BME） during neural differentiation of mesenchymal stem cells （MSCs） in vitro...Objective To study the protective effects of hydroxysafflor yellow A （HSYA） against the oxidative damage caused by β-mercaptoethanol （BME） during neural differentiation of mesenchymal stem cells （MSCs） in vitro. Methods When the confluence reached 50%-60%, 4th passage MSCs were divided into three groups to culture. Gt ： normal group which was cultured using basic medium （DMEM containing 10% FBS all the time）; G2： unprotected group which was continuously cultured using basic medium for 24 h, and then cultured using pre-induction medium （DMEM containing 10% FBS and 1 mmol/L BME）; G3： protected group which was firstly cultured using protective medium （DMEM containing 10% FBS and 160 mg/L HSYA） for 24 h, and then cultured using pre-induction medium for 24 h. After these treatments as above, cell viability, relative levels of SOD/GSH and apoptosis rate were respectively detected. The expression of Bcl and Bax was examined by Western blotting. After HSYA protection and BME pre-induction, neural induction was performed. The expression of NSE and MAP-2 was respectively analyzed on cellular and molecular levels. Results Compared with unprotected group, 160 mg/L HSYA could obviously improve cells viability, maintain high level of SOD and GSH in MSCs, reduce apoptosis rate and improve the ratio of Bcl/Bax. After protection with 160 mg/L HSYA, the survival time of neuron-like cells could be extended. Immunocytochemical staining showed that after 10 h of neural induction, the differentiated neuron-like cells in protected group were still in a good state, and the mRNA levels of NSE and MAP-2 were increased during the induction course checked. Conclusion HSYA could improve the resistance of cells to the oxidative damage caused by BME.展开更多
文摘Objective:To investigate the mechanism by which hydroxyl safflower yellow A,an active component of safflower (Carthamus tinctorius L.),promotes apoptosis in abnormal human umbilical vein endothelial cells (HUVECs).Methods:Supernatant of BGC-823 was used to stimulate HUVECs to establish a model of abnormal proliferation of HUVECs.After determining an ideal concentration of HSYA by MTT assay,apoptosis was detected with flow cytometry and TUNEL assay.Mechanism of apoptosis was assessed using quantitative real-time polymerase chain reaction,Western blot,and ELISA.Results:A range of concentrations of HSYA inhibited proliferation and promoted apoptosis of abnormal HUVECs.As the rate of apoptosis increased,mRNA expression of caspase-3 increased while expression of mutant p53 decreased.HSYA had no effect on Fas gene expression.Analogously,protein expression of Bax was increased while those of Bcl-2,Fas,and Fas-L were decreased.Conclusions:HSYA appears to induce apoptosis of HUVECs with the stimulation of the supematant of tumor cells.The mechanism of apoptosis by HSYA may involve activation of the mitochondrial apoptotic pathway and regulation of the expressions of Bcl-2,Bax,and p53.
基金Major Projection of Hebei North University(No.ZD201413)
文摘Objective To study the protective effects of hydroxysafflor yellow A （HSYA） against the oxidative damage caused by β-mercaptoethanol （BME） during neural differentiation of mesenchymal stem cells （MSCs） in vitro. Methods When the confluence reached 50%-60%, 4th passage MSCs were divided into three groups to culture. Gt ： normal group which was cultured using basic medium （DMEM containing 10% FBS all the time）; G2： unprotected group which was continuously cultured using basic medium for 24 h, and then cultured using pre-induction medium （DMEM containing 10% FBS and 1 mmol/L BME）; G3： protected group which was firstly cultured using protective medium （DMEM containing 10% FBS and 160 mg/L HSYA） for 24 h, and then cultured using pre-induction medium for 24 h. After these treatments as above, cell viability, relative levels of SOD/GSH and apoptosis rate were respectively detected. The expression of Bcl and Bax was examined by Western blotting. After HSYA protection and BME pre-induction, neural induction was performed. The expression of NSE and MAP-2 was respectively analyzed on cellular and molecular levels. Results Compared with unprotected group, 160 mg/L HSYA could obviously improve cells viability, maintain high level of SOD and GSH in MSCs, reduce apoptosis rate and improve the ratio of Bcl/Bax. After protection with 160 mg/L HSYA, the survival time of neuron-like cells could be extended. Immunocytochemical staining showed that after 10 h of neural induction, the differentiated neuron-like cells in protected group were still in a good state, and the mRNA levels of NSE and MAP-2 were increased during the induction course checked. Conclusion HSYA could improve the resistance of cells to the oxidative damage caused by BME.
