| Objective:The Callicarpa nudiflora hook is Verbenacase and Callicarpa genus plant, Hainan authentic medicinal materials, the leaves is main medicinal parts, which has detoxification sore cure and convergence bleeding and dispels wind and dampness, Can be used to treat burns, burns and trauma hemorrhage, etc. Existing studies have shown that Callicarpa nudiflora hook extract can promote bleeding, inhibition of thrombosis, inhibit bacterial and cytotoxicity, resistance to inflammation, strengthens the immune and other pharmacological activity. The paper main purpose is determine effective parts Callicarpa nudiflora hook Hemostatic activity, and initial exploration of its Hemostatic activity and mechanism of action from the perspective of platelet activation. Application of the broken tail method and Tajima improved method to examine the Callicarpa nudiflora Hook extracts on bleeding time and clotting time in mice. Methods:(1) The four parts from Callicarpa nudiflora hook petroleum ether, ethyl acetate, n-butanol, water were batches tested, take the same batch of 50 mice were randomly divided into control group, positive group and Yunnan Baiyao group and Callicarpa nudiflora hook extracts high dose group, middle dose group and low dose group of five groups, based on body weight dose administered in vivo continuous 3d, determine the effective area.(2) On this basis, using rat, grouping up, medication for ten days, the Callicarpa nudiflora Hook ethyl acetate and n-butanol extract prothrombin time, fibrinogen, activated partial thromboplastin time, thrombin time were test.(3) Then respectively platelet count and micro plate to observe the number and activity of Callicarpa nudiflora Hook extract on rat platelets, and to observe the content TAT, SELP, TXB2, 6-K-PG of Callicarpa nudiflora Hook extract on rat platelets.(4) Finally, using the Western blot assay influence Callicarpa nudiflora Hook extract on platelet AKT, P38 MAPK, ERK and other signal transduction molecules expressed, clear Hemostatic mechanism. Results:(1) Callicarpa nudiflora Hook petroleum ether and water extract part is not affect of the BT and CT in mice, there was no significant difference(P > 0.05), and ethyl acetate and n-butanol extract part have a certain impact of BT and CT in mice. Comparison with the control group, Yunnan Baiyao positive group and different doses of ethyl acetate extraction section could significantly shorten BT(P < 0.01) and CT(P < 0.05) in mice. And ethyl acetate extraction section similar role, different doses of n-butanol extraction section may also significantly shorten BT(P < 0.01) and CT(P < 0.05) in mice.(2) In coagulation experiments Callicarpa nudiflora Hook ethyl acetate and n-butanol extract PT, FIB, TT had no significant effect(P > 0.05), but can reduce the APTT, and a significant statistically significant difference(P < 0.05).(3) It was found in platelets, compared with the control group, Callicarpa nudiflora Hook ethyl acetate and n-butanol extract could significantly increase test animals platelet count(P < 0.05); It was found that platelet aggregation in ADP(concentration of 5.46 mmol / L) induced, Callicarpa nudiflora Hook ethyl acetate and n-butanol extract group increased aggregation rate, compared with the control group, the aggregation rate was significant difference(P < 0.05); Platelet TAT, TXB2, 6-K-PG, SELP content detection found: these two extracts TAT had no significant effect on the platelets(P > 0.05), but can be increased platelet TXB2, 6-K-PG, SELP content, and there is significant difference(P < 0.05).(4) In the Western blot test, Callicarpa nudiflora Hook ethyl acetate extract, n-butanol extract could significantly stimulate Akt, p-Akt expression of blank group Akt, p-Akt relative optical density and the β-actin values were increased, but has little effect on the other two proteins. Conclusion:(1) Callicarpa nudiflora Hook ethyl acetate and n-butanol can shorten BT and CT, reduces APTT, there Hemostatic activity;(2) Callicarpa nudiflora Hook ethyl acetate and n-butanol can increase the number of platelets and stimulate platelet activation;(3) Hemostatic activity Callicarpa nudiflora Hook ethyl acetate and n-butanol under P2Y12 receptor may signal transduction by affecting the surface of platelets in platelet ADP. |
