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目的:研究裸花紫珠(Callicarpa nudiflora)正丁醇提取物的止血活性及其可能的作用机制。方法:雌性KM小鼠50只,随机分为空白对照组、云南白药组(1.33g/kg)、裸花紫珠低(0.188 g提取物/kg)、中(0.375g提取物/kg)、高剂量组(0.750g提取物/kg),连续给药3d后测定受试动物的出血时间(Bleeding time,BT)和凝血时间(Clotting time,CT)。取雄性SD大鼠50只,随机分为空白对照组、云南白药组(0.93 g/kg)、裸花紫珠低(0.131g提取物/kg)、中(0.261g提取物/kg)、高剂量组(0.523g提取物/kg),连续给药10d。给药结束后分别测定血小板(Platelet,PLT)计数、二磷酸腺苷(adenosine diphosphate,ADP)诱导的血小板聚集以及血小板中P-选择素(P-Selectin)含量,观察裸花紫珠提取物对血小板数量及活性的影响。采用Western Blot法,检测裸花紫珠提取物对血小板中p-Akt表达的影响。结果:给药后,裸花紫珠正丁醇提取物可显著缩短小鼠的出血时间和凝血时间。与正常组相比,0.131、0.523g裸花紫珠提取物/kg BW均可显著增加大鼠血小板计数。裸花紫珠正丁醇提取物可显著促进ADP诱导的血小板聚集;进一步研究表明,裸花紫珠提取物可显著增加血小板内P-selectin的表达,并促进Akt蛋白的磷酸化。结论:裸花紫珠提取物具有明显的止血作用,促进血小板PI3K/Akt信号转导、刺激血小板的活化可能是其止血作用的机制之一。
Objective: To study the hemostatic activity of n-butanol extract of Callicarpa nudiflora and its possible mechanism. Methods: Fifty female KM mice were randomly divided into three groups: blank control group, Yunnan Baiyao group (1.33g / kg), naked purple flower beads (0.188g extract / kg) High dose group (0.750 g extract / kg). After 3 days of continuous administration, the bleeding time (Bleeding time, BT) and clotting time (CT) of the animals were measured. Fifty male Sprague-Dawley rats were randomly divided into 4 groups: blank control group, Yunnan Baiyao group (0.93 g / kg), naked purple flower beads (0.131 g extract / kg) Dose group (0.523 g extract / kg) for 10 days. At the end of the treatment, the Platelet (PLT) counts, the platelet aggregation induced by adenosine diphosphate (ADP) and the P-selectin content in platelets were measured. Effect of Platelet Number and Activity. Western Blot method was used to detect the expression of p-Akt in platelets of Bamei purple beads extract. Results: After administration, the n-butanol extract of Balsam pear significantly reduced the bleeding time and clotting time of mice. Compared with the normal group, 0.131, 0.523g naked flower purple beads extract / kg BW can significantly increase rat platelet count. Bare n-butanol extracts of naked flowers could significantly promote platelet aggregation induced by ADP. Further studies showed that BMCs significantly increased the expression of P-selectin in platelets and promoted the phosphorylation of Akt protein. CONCLUSION: The naked flower extract has obvious hemostatic effect, promoting platelet PI3K / Akt signal transduction and stimulating platelet activation may be one of its mechanisms of hemostasis.