叠鞘石斛提取物抗血小板聚集作用研究

Study on the activity of anti-platelet aggregation of Dendrobium denneanum

目的 观察叠鞘石斛提取物的抗血小板聚集活性,探讨叠鞘石斛的抗血栓作用.方法 将21只大鼠按体重随机分为提取物1组、提取物2组、提取物3组、提取物4组、提取物5组、提取物6组及阳性对照组,每组3只;同样将9只豚鼠按体重随机分为提取物5组、提取物6组及阳性对照组,每组3只.各组大鼠分别采用二磷酸腺苷（adenosine diphosphate,ADP）和花生四烯酸（arachidonic acid,AA）诱导大鼠和豚鼠体外血小板聚集实验,收集富血小板血浆（plate let rich plasma,PRP）.各组PRP分别加入不同浓度的叠鞘石斛提取物1、2、3、4、5、6或阿司匹林进行干预,检测5 min内最大聚集率,并计算血小板聚集的抑制率和各受试物的半数抑制浓度（IC50）.结果 与空白对照组比较,12.399 mmol/L提取物1组,8.960 mmol/L提取物2组,17.505 mmol/L提取物3组,7.836、3.918 mmol/L提取物5组,0.755、0.377 mmol/L提取物6组对ADP诱导的血小板最大聚集率[（32.57±0.28）%比（45.20±2.64）%]、[（16.66±3.67）%比（39.37±8.05）%]、[（0.00±0.00）%比（32.52±3.05）%]、[（1.11±1.93）%、（7.49±9.47）%比（43.54±10.68）%]、[（8.66±7.50）%、（22.71±8.24）%比（40.65±6.61）%]降低（P〈0.05或P〈0.01）,提取物2、3、5、6的IC50分别为4.951、3.808、1.699、0.391 mmol/L;与空白对照组比较,7.836 mmol/L提取物5组、0.377 mmol/L提取物6组对AA诱导的血小板最大聚集率[（0.00±0.00）%比（52.97±5.49）%]、[（0.00±0.00）%比（53.47±6.04）%]降低（P〈0.05或P〈0.01）,提取物5、6的IC50分别为3.143、0.100 mmol/L.结论 叠鞘石斛多种成分具有抗血小板聚集作用,在血栓性疾病中具有一定的积极作用.

Objective To study the anti-platelet aggregation activity of Dendrobium denneanum. Methods A total of 21 rats were randomly divided into the extract 1 group, the extract 2 group, the extract 3 group, the extract 4 group, the extract 5 group, the extract 6 group and the positive control group, with 3 in each group. While 9 guinea pigs were randomly divided into the extract 5 group, the extract 6 group and the positive control group, 3 guinea pigs per group. The platelet aggregation was induced by adenosine triphosphate （ADP） in vitro in rats and arachidonic acid （AA） in vitro in guinea pigs. Plate let rich plasma （PRP） was collected. Each group of PRP was added with different concentrations of Dendrobium denneanum extract 1, 2, 3, 4, 5, 6 or aspirin to intervene. The maximal aggregation rate were detected in 5 min, and to calculate the inhibition rate of platelet aggregation and half of each test substance inhibitory concentration （IC50）. Asp was detected in 5 min. The above steps were repeated three times. The inhibition rate of platelet aggregation and the IC50 of each subject were calculated. The maximum aggregation rate of the drug delivery group and the blank control group was compared to evaluate the antithrombotic activity of the Dendrobium with two different induction methods. Results Compared with the blank control group, the maximal aggregation rate induced by ADP of the extract 12.399 mmol/L dose group, the extract 28.96 mmol/L dose group, the extract 317.505 mmol/L dose group , the extract 57.836, 3.918 mmol/L dose group, the extract 60.755, 0.377 mmol/L dose group （32.57% ± 0.28% vs. 45.20% ± 2.64%）, （16.66% ± 3.67% vs. 39.37% ± 8.05%）, （0.00% ± 0.00% vs. 32.52% ± 3.05%）, （1.11% ± 1.93%, 7.49% ± 9.47% vs. 43.54% ± 10.68%）, （8.66% ± 7.50%, 22.71% ± 8.24% vs. 40.65% ± 6.61%） significantly decreased （P〈0.05 or P〈0.01）. Compared with the blank control group, the maximal aggregation rate induced by AA of the extract 57.836 mmol/L dose group, the extract 60.377 mmol/L dose group （0.00% ± 0.00% vs. 52.97% ± 5.49%）, （0.00% ± 0.00% vs. 53.47% ± 6.04%） significantly decreased （P〈0.05 or P〈0.01）. The IC50 of extracts 5 and 6 were 3.143 and 0.100 mmol/L, respectively. Conclusions There were many kinds of ingredients in Dendrobium denneanum which had anti-platelet aggregation activity. Dendrobium denneanum had positive effects on thrombotic diseases.