ABSTRACT
Objective To study the effects of abnormal blood flow on the secretion of ET-1/NO and the expression of the mRNA and the protein of ET-1, eNOS, VCAM-1, ICAM-1 and MCP-1 in human umbilical vein endothelial cells (HUVECs), so as to explore the mechanism of atherosclerosis （AS） caused by abnormal hemodynamics. MethodsThe HUVECs were divided into stress group, wall pressure group and normal group according to the different stress. The HUVECs were cultured under the corresponding stress for 24 hours and then collected. The secretion levels of NO and ET-1 were detected by enzyme method and radioimmunoassay method. The mRNA expression levels of eNOS and ET-1 were detected by qPCR. The expression levels of the mRNA and the protein of VCAM-1, ICAM-1, MCP-1 were detected by qPCR and Western blot. Results Compared with normal group, the secretion level and the mRNA expression level of ET-1 in wall pressure group increased significantly (P＜0.01), and the secretion level of NO and the mRNA expression level of eNOS in stress group also increased significantly (P＜0.01), The expressions level of the mRNA and the protein of VCAM-1, ICAM-1 and MCP-1 obviously increased in stress group and wall pressure group (P＜0.01). Conclusions Stress or wall pressure acting on HUVECs alone could lead to its dysfunction of the secretion and the expression of gene and protein. The mechanism of AS caused by abnormal blood flow was related to these dysfunction of HUVEC.
ABSTRACT
Objective To study the effects of abnormal blood flow on the secretion of ET-1/NO and the expression of the mRNA and the protein of ET-1, eNOS, VCAM-1, ICAM-1 and MCP-1 in human umbilical vein endothelial cells (HUVECs), so as to explore the mechanism of atherosclerosis （AS） caused by abnormal hemodynamics. MethodsThe HUVECs were divided into stress group, wall pressure group and normal group according to the different stress. The HUVECs were cultured under the corresponding stress for 24 hours and then collected. The secretion levels of NO and ET-1 were detected by enzyme method and radioimmunoassay method. The mRNA expression levels of eNOS and ET-1 were detected by qPCR. The expression levels of the mRNA and the protein of VCAM-1, ICAM-1, MCP-1 were detected by qPCR and Western blot. Results Compared with normal group, the secretion level and the mRNA expression level of ET-1 in wall pressure group increased significantly (P＜0.01), and the secretion level of NO and the mRNA expression level of eNOS in stress group also increased significantly (P＜0.01), The expressions level of the mRNA and the protein of VCAM-1, ICAM-1 and MCP-1 obviously increased in stress group and wall pressure group (P＜0.01). Conclusions Stress or wall pressure acting on HUVECs alone could lead to its dysfunction of the secretion and the expression of gene and protein. The mechanism of AS caused by abnormal blood flow was related to these dysfunction of HUVEC.
ABSTRACT
Objective To investigate the characteristics of growth, accumulation of indole alkaloid, and the optical concentration of the nutrition components in the Catharanthus roseus mutant cells induced by the colchicine, looking forward to getting the ideal material producing alkaloids which was suited for industrialized cell cultivation. Methods Mutant cells cultivated on the MS medium were harvested according to experiment designs,and the fresh weight was got.The indole alkaloid as extracted by organic solvent. The ajmalicine and catharanthine extracted from the samples were analyzed by RP-HPLC. Results The growth rate was faster and the indole alkaloid accumulation was more when the mutant cells were cultivated generations 30 and the both were sharply decreased at generations 45, while the contents of medical compounds amounted to the peak at the generations 20. The content of medical components and alkaloid accumulation were evidently prompted by the tryptophan added in the media, especially at the concentration of Ca2+ 1 760 and Zn2+ 12.6 mg/L, respectively. Conclusion C.roseus mutant cells probably is an ideal material for industrialized cell cultivation.