银杏内酯B抑制血小板与肿瘤细胞相互作用的实验研究  

Experimental Study on Inhibition of Interaction between Platelets and Tumor Cells by Ginkgolide B

作　　者：薛昌雯 邵小宝 陈鑫[1] 周琳 朱培元 XUE Changwen;SHAO Xiaobao;CHEN Xin;ZHOU Lin;ZHU Peiyuan(Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine,Nanjing 210022)

机构地区：[1]南京中医药大学附属南京中医院输血科 [2]南京市浦口区中医院肛肠科,江苏南京210022

出　　处：《临床输血与检验》2024年第2期205-212,共8页Journal of Clinical Transfusion and Laboratory Medicine

基　　金：南京市卫生科技发展专项资金项目(No.YKK21195);江苏省输血协会英科新创科研基金(No.JS2022006)资助。

摘　　要：目的研究银杏内酯B(ginkgolide B,GB)对血小板活化的抑制作用,以及基于该抑制作用对血小板和肠癌HCT-116细胞间作用的影响。方法采用不同浓度GB(5、15、30μmol/L)处理健康志愿者全血或血小板,通过血栓弹力图(Thromboelastogram,TEG)检测GB对血小板二磷酸腺苷(Adenosine diphosphate,ADP)、花生四烯酸(arachidonic acid,AA)途径的抑制率,采用流式细胞术检测GB对凝血酶和ADP激活血小板后CD62P和PAC-1表达的影响,通过细胞划痕试验观察GB对血小板与HCT-116细胞作用24、48 h后细胞迁移的影响,通过细胞黏附试验观察GB对血小板与HCT-116细胞黏附情况的影响。结果(1)TEG试验结果显示,随着GB浓度增加,AA抑制率均显著增加且呈剂量依赖(P<0.001),与对照组相比差异均有统计学意义(P<0.001);药物组与空白对照组相比,ADP抑制率均显著增加且呈剂量依赖,差异均有统计学意义(P<0.001)。(2)流式细胞术检测结果显示,凝血酶和ADP激活血小板后药物组的CD62P与PAC-1阳性率随着药物浓度升高而降低且呈剂量依赖性(P<0.001),凝血酶激活途径的药物5μmol/L组的PAC-1阳性率低于对照组,差异无统计学意义(P>0.05),ADP激活途径的药物5μmol/L组的CD62P阳性率、5μmol/L和15μmol/L组的PAC-1阳性率均低于对照组,差异无统计学意义(P>0.05),余各浓度组与对照组相比差异均有统计学意义(P<0.01)。(3)划痕试验结果显示,不同浓度药物组的24、48 h细胞迁移率分别较凝血酶组与ADP组降低,且差异有统计学意义(P<0.01),15μmol/L组的24、48 h细胞迁移率明显低于5μmol/L组,差异具有统计学意义(P<0.05),30μmol/L组迁移率低于15μmol/L组,但差异无统计学意义(P>0.05)。(4)黏附试验结果显示,活化的血小板可增强其与肿瘤细胞之间的黏附作用,但药物组各组的黏附率均低于阳性组,且随着药物浓度增加黏附率随之降低。结论GB可通过AA途径及ADP途径抑制血小板活化,进而抑Objective To study the inhibitory effect of ginkgolide B(GB)on platelet activation as well as on the interaction between platelets and colorectal cancer HCT-116 cells.Methods Whole blood or platelets were treated with different concentrations of GB(5,15,30μmol/L).Thromboelastogram(TEG),then platelet Adenosine diphosphate was detected.The inhibitory rate of ADP and arachidonic acid(AA)pathway was detected by flow cytometry.The effects of GB on the expression of CD62P and PAC-1 after thrombin and ADP-activated platelets were detected.The effect of GB on the cell migration of platelets and HCT-116 cells after 24 and 48 h interaction was observed by cell scratch test,and the effect of GB on the adhesion of platelets to HCT-116 cells was observed by cell adhesion test.Results(1)TEG test results showed that with the increase of GB concentration,AA inhibition rates increased significantly in dose-dependent manner(P<0.001),the difference was statistically significant compared with the control group(P<0.001);The inhibition rate of ADP in the drug group was significantly increased compared with the control group in a dose-dependent manner(P<0.001),the differences were statistically significant(P<0.001).(2)Flow cytometry showed that after thrombin and ADP activated platelets,the positive rates of CD62P and PAC-1 in the drug group decreased with the increase of drug concentration in a dose-dependent manner(P<0.001),the positive rate of PAC-1 in 5µmol/L group with thrombin activation pathway was lower than that in control group,and the difference was not statistically significant(P<0.05),the positive rate of CD62P in 5µmol/L group,the positive rate of PAC-1 in 5µmol/L and 15µmol/L groups of ADP-activated pathway drugs was lower than that of control group,and the difference was not statistically significant(P<0.05),and there were statistically significant differences between the remaining concentration groups and the control group(P<0.01).(3)Scratch test results showed that 24 and 48 h cell mobility in different concentra

关 键 词：银杏内酯B 结直肠癌 血小板 黏附 迁移 

分 类 号：R285[医药卫生—中药学]