Effective Mechanism Of Compound Ligusticum Chuanxiong And Angelica Sinensis On Vascular Cells

To better understand the complex substance and effective mechanism of Traditional Chinese Medicine, in this paper, the molecular mechanisms of compound Ligusticum chuanxiong and Angelica sinensis and its active substances were estimated on the vascular smooth muscle cell and endothelial cell involved in the ROS, MAPK signal pathways and the relation of the signal molecules. The biological effect of the extract Ligusticum chuanxiong and Angelica sinensis (ELCAS) on vascular smooth muscle cell proliferation and endothelial cell damage induced respectively by serum and H2O2 was estimated. The data show that ELCAS significantly inhibited vascular smooth muscle cell proliferation and endothelial cell damage, ELCAS suppressed the intracellular ROS formation, resulting in inactivation of ERK, JNK, p38, and increment of p21 level, decrement of the Cyclin D1 and pRb, subsequently, inhibited the proliferation of vascular smooth muscle cell. The analysis of HPLC suggests ELCAS contained ferulic acid, essential oil, tetramethlpyrazine. The cell proliferation and damaged models were used to assay the three kinds of active substances. The results suggest that the pharmacological effect of ferulic acid was better than others. Serum significantly activated ERK pathway, but ferulic acid inhibited activation of ERK and peaked at 10 min, suggesting that the inhibition of ferulic acid on the endothelial cell proliferation was involved in the ERK pathway. On the other hand, ferulic acid increased the NO production and peaked at 4 h. SNP, NO donor, decreased activation of ERK. PD 98059 inhibited the activation of ERK, but had no effect on NO production, suggesting NO down-regulated the activation of ERK. Angiotensin II significantly stimulated the vascular smooth muscle cell proliferation, but ferulic acid inactivated angiotensin II induced activation of ERK, JNK rather than p38, and upregulated the expression of p21, suppressed Cyclin D1 and phosphorylation of pRb, resulting in the cell arrested in G₀/G₁. To further understand the relation of experiment and model, the signal pathway model was built. The model may be a new idea for analysis of pharmacological effect. The signal pathway model suggests that AT₁ receptor was activated quickly and peaked at 1 minute, and the phosphorylation of RasGTP, Raf*, MEKPP peaked at 5 minute. ERK was activated at 15 minute, but it was inhibited by ferulic acid, the experimental results were also in line with the model, the correlation coefficient was 99.3%. Conclusion, it is the first time to investigate the molecular mechanism of compound Ligusticum chuanxiong and Angelica sinensis and its active substances on the effect of vascular cells, which was associated with the inactivation of ROS and MAPK signal pathway.