Compound Danshen Dropping Pill Regulate The Expression Of MicroRNAs In Mice

Compound danshen dropping pill(DSP) is composed of salvia miltiorrhiza, notoginseng and borneol. The DSP is used for coronary heart disease, angina pectoris as well as hypertension, hyperlipidemia, cerebral thrombosis. Pharmacological studies have shown that phenolic acids in DSP could increase the activity of anticoagulant and fibrinolysis, inhibit platelet aggregation and thrombosis, oxidation and protect cardiac microvascular endothelial cells, improve microcirculation and so on; Saponins could improve the function of endothelial cells, inhibit proliferation of vascular smooth muscle cell, resist thrombosis, dilate blood vessels and protect myocardial.In order to understand the mechanisms of the therapeutic effect of compound danshen dripping pills(DSP) on cardiovascular disease, our study aims to consider the expression effect of CDDP on mouse micro RNA. Through the deeply research of differential miRNAs' on cardiovascular disease, we expect to reveal the molecular mechanisms of CDDP on cardiovascular disease through a new perspective.To find the differential endogenous miRNAs in mice caused by CDDP, the Institute for Cancer Research(ICR)mice were eaten for 500 ul 4.4mg/ml DSP solution as the experimental group and for equivalent water as the control group, then took blood, heart, liver, kidney, lung and stomach respectively at the time of 0h, 0.5h, 1h, 3h, 6h and 12 h after the mice was performed the stomach perfusion. The s RNA sequencing analysis in blood showed that there were 42 differential expression miRNAs including,16 up-regulated miRNAs and 26 down-regulated miRNAs. Further verified by quantitative reverse-transcription PCR(q RT-PCR), 12 miRNAs are up-regulated, 1 miRNAs are down-regulated and 3 miRNAs have no obvious change among the 16 RNA-sequenced up-regulated miRNAs. And there are 20 down-regulated miRNAs, 1 up-regulated miRNA and 5 no obvious change miRNAs among the 26 RNA-sequenced down-regulated miRNAs. In the mice heart, 11 miRNAs show up-regulated and 5 miRNAs have no obvious change among 16 RNA-sequenced up-regulated miRNAs, and among 26 RNA-sequenced down-regulated miRNAs, 21 miRNAs show up-regulated and other 5 miRNAs have no obvious change.In order to verify the roles of these differential miRNAs in myocardial cell apoptosis and hypertrophy, we use different myocardial cell hypertrophy inducer Phenylephrine(PE,10? M), Angiotensin?(Ang ?,10? g/L)and insulin-like growth factors-1(IGF-1,100 n M) to induce hypertrophy of primary cultured myocardial cell and test the expression of 42 miRNAs.The results indicate that the expression of 19 miRNAs change differentially when treated by PE, 6 miRNAs have an obviously change when treated by Ang?, and the other miRNAs change obviously when treated by IGF-1. miR7084-5p is up-regulated by PE and Ang? and down-regulated by IGF-1.In order to explore the function of these differential miRNAs in myocardial cell apoptosis and hypertrophy, 5 miRNAs(miR-106a-5p?miR-1964-3p?miR-467d-3p?miR-468-3p and miR-672-5p) were chosen to be verified and found that the inhibition of miR106a-3p?miR467d-3p and miR486-3p can obviously reduce the diameter of hypertrophic myocardial cell. Then cell viability was detected by MTT method and the results showed that the overexpression of miR106 a had an inhibitory effect on the cell apoptosis, and the cell apoptosis reduced when the expression of miR672-5p was inhibited.Based on the above results, we speculate that CDDP probably regulate the myocardial hypertrophy by the regulation of miR106a-5p, miR467d-3p and miR486-3p, and play a role in the cell apoptosis through the regulation of miR106a-5p and miR672-5p.