RESUMO
Objective: In this study, we examined whether Danhong Injection (DHI) exerted a relaxation effect in vascular smooth muscle cells (A7r5) in co-culture system by human umbilical vein endothelial cells (Ea.Hy926) and studied its mechanisms. Methods: First of all, Western blotting assay was carried out to quantify the protein expression of MLC and P-MLC in A7r5 cells. Secondly, calcium assay kit was used to detect the changes in intracellular calcium while the changes of CaM mRNA expression were subsequently detected by RT-PCR. MTT assay was used to detect the effect of DHI in proliferation of A7r5 cell, which is induced by PDGF-BB. Finally, the co-culture system of EA.Hy926 and A7r5 cells was constructed by a Transwell. After the treatment of Hy926 cells with DHI for co-culture system, the protein expression of MLC and P-MLC in A7r5 cells in co-culture system was also quantified by Western blotting. Cell-based ELISA was used to observe the cAMP generation in A7r5 cells. Results: DHI (10 μL/mL) contributed to the decrease of the ratio of P-MLC/MLC (P < 0.05) and CaM mRNA expression (P < 0.01) in A7r5 cells, which are cultured individually, but had no significant effect in intracellular calcium concentration and the expression of MLC proteins. In co-culture system, DHI (10 μL/mL) could still decrease the ratio of P-MLC/MLC (P < 0.05) and increase the secretion of cAMP (P < 0.01) in A7r5 cells. In addition, DHI contributed to suppress the proliferation of A7r5 cells which were induced by the PDGF-BB (P < 0.01). Conclusion: DHI can directly relax vascular smooth muscle cells by reducing the ratio of P-MLC/MLC and the expression of CaM mRNA, meanwhile indirectly relaxing the vascular smooth muscle cells in co-culture system by promoting the secretion of cAMP which is increased by vasodilatory factors in endothelial cells.
RESUMO
Cardiovascular disease is becoming the leading cause of death in the world. Salvia miltiorrhiza has a history of thousands of years in clinical therapy in China, which is widely used in the treatment of coronary heart disease, myocardial infarction, blood circulation diseases, and other cardiovascular diseases. More and more studies show that danshensu, the main water-soluble component in S. miltiorrhiza, has a variety of pharmacological effects, including inflammation and tumor inhibition, neuro and myocardial protection, immunity improving, and so on. With further research, the mechanisms of these pharmacological effects have been gradually clear. In this paper, we focus on the mechanism of danshensu on cardiovascular protection and provide a theoretical basis for the further study and clinical application of danshensu. The aspects of mechanism include inhibition on ischemia/reperfusion injury, hypertension, myocardial infarction, atherosclerosis, high cholesterol, and hypertension, protection of endothelial cells, and so on.