侧柏叶提取物对血液凝固的影响及其机制

The effect of platycladus orientalis leaf extract on blood coagulation and its mechanisms

目的探究侧柏叶提取物对血液凝固的影响及其机制。方法制备侧柏叶水提物及其石油醚、乙酸乙酯和正丁醇提取物,取家兔颈总动脉血液,用凝血板法,观察侧柏叶水提物和石油醚、正丁醇及乙酸乙酯提取物对血液凝固的影响;以RT、APTT、PT以及TT为指标,观察体外实验侧柏叶提取物对血液凝固的影响。将健康昆明种小鼠随机分为侧柏叶水提物低、高浓度组,乙酸乙酯提取物低、高浓度组,正丁醇提取物低、高浓度组以及空白对照组,每组8只;以凝血时间、APTT、PT、TT及凝血因子Ⅻ为指标,观察侧柏叶提取物对血液凝固的影响。通过灌胃5ml/kg的无水乙醇制备胃溃疡出血模型,以胃出血面积、胃组织切片后胃黏膜细胞形态为指标,观察侧柏叶提取物对胃出血模型的作用。结果在体外实验与空白对照组相比,石油醚、乙酸乙酯、正丁醇提取物和水提物组显著延长凝血时间和RT(P<0.05);正丁醇提取物、水提物组明显延长APTT和PT(P<0.05),乙酸乙酯提取物明显延长PT(P<0.05)。在体内实验与空白对照组相比,乙酸乙酯、正丁醇提取物、水提物低浓度组凝血时间明显缩短(P<0.05);乙酸乙酯提取物高浓度组和正丁醇提取物低浓度组分别显著缩短APTT和PT(P<0.05),而正丁醇、水提物高浓度组则显著延长TT(P<0.05);与空白对照组相比,乙酸乙酯低浓度组、正丁醇提取物高、低浓度组血浆ⅩⅡ因子含量明显升高(P<0.05)。与模型组相比,乙酸乙酯提取物高、低浓度组、正丁醇提取物高、低浓度组和水提物高、低浓度组胃出血面积明显缩小(P<0.05),乙酸乙酯提取物低浓度组胃出血面积最小。结论侧柏叶体内实验可促进凝血,乙酸乙酯提取物是主要的止血提取物,且低浓度的乙酸乙酯提取物止血效果更好。

Objective To investigate the effect of Platycladus orientalis leaves on blood coagulation and its related mechanisms.Methods In vitro coagulation experiment:prepared the extract of Platycladus orientalis leaves by petroleum ether,ethyl acetate,n-butanol and water.Took the blood from the common carotid artery of rabbits,and observed the effect of the extract of Platycladus orientalis leaves by water,petroleum ether,n-butanol and ethyl acetate on blood coagulation with the clotting plate method.Using RT,APTT,PT,and TT as indicators,observed the effects of Platycladus orientalis leaves and its extracts on blood coagulation in vitro experiments.In vivo coagulation experiment:Healthy Kunming mice were randomly divided into low and high concentration groups of water extract from Platycladus orientalis leaves.Low and high concentration groups of ethyl acetate extract.Low and high concentration groups of n-butanol extract,as well as blank group.Using coagulation time,APTT,PT,TT,and coagulation factor XIII as indicators,the effects and related mechanisms of Platycladus orientalis leaves and its extracts on blood coagulation were studied through in vivo experiments.The effect of extract of Platycladus orientalis leaves on gastric ulcer bleeding model:A gastric ulcer bleeding model was prepared by gavage with 5ml/kg anhydrous ethanol.The effect of Platycladus orientalis leaves and its extract on gastric bleeding model was observed using gastric bleeding area and gastric mucosal cell morphology after gastric tissue sectioning as indicators.Results In vitro experiment:Compared with the blank control group,petroleum ether,ethyl acetate,n-butanol extract,and water extract significantly prolonged coagulation time and RT(P<0.05);The n-butanol extract,water extract significantly prolonged APTT and PT(P<0.05),and ethyl acetate extract significantly prolonged PT(P<0.05).In vivo experiment:Compared with the blank control group,the low concentration group of ethyl acetate,n-butanol and water extract significantly shortened the coagulation time(P<0.05);The high concentration group of ethyl acetate extract and the low concentration group of n-butanol extract significantly shortened APTT and PT separately(P<0.05),while the high concentration group of n-butanol and water extract significantly prolonged TT(P<0.05);Compared with the blank control group,the low concentration group of ethyl acetate,the high and low concentration group of n-butanol extract showed a significant increase in plasma factor XIII content(P<0.05).Compared with the model control group,the areas of gastric bleeding in the high and low concentration groups of ethyl acetate extract,high and low concentration groups of n-butanol extract,and high and low concentration groups of water extract were significantly reduced(P<0.05).The low concentration group of ethyl acetate extract had the smallest area of gastric bleeding.Conclusion In vivo experiments,Platycladus orientalis leaves mainly promote coagulation,with ethyl acetate as the main hemostatic site,and low concentrations of ethyl acetate extract have better hemostatic effects.