王珺 张雅月 王佳 王冲 陈科 陈信义 郎海燕

摘要 目的：基于血管活性因子特性探討“从脾论治”ITP疗效机制。方法：以被动型免疫造模法建立ITP小鼠模型，以强的松、健脾益气方、健脾益气摄血方、归脾汤为干预药物，酶联免疫吸附法（ELISA）检测模型小鼠血清ET-1、NO、NOS3、TXA2、PGI2、vWF、VCAM-1、TM含量。结果：1）升血小板疗效：与正常对照组比较，注射APS后各组小鼠外周血PLT计数明显下降（P<0.01）;与模型对照组比较，给药第8天各组小鼠外周血PLT计数明显升高（P<0.01）。2）止血疗效：给药前除正常对照组外，实验各组均有不同程度出血倾向;给药第6天、第8天，与模型对照组比较，实验各组出血分级下降（P<0.01）。3）血管促凝因子：与正常组比较，各组ET-1检测值明显下调（P<0.01）;与正常对照组、模型对照组比较，实验各组TXA2检测值明显上调（P<0.01）;与正常组比较，实验各组vWF检测值明显下调（P<0.01）;与正常对照组比较，实验各组VCAM-1检测值明显下调（P<0.01）。4）血管抗凝因子：与正常组比较，实验各组NO检测值明显下降（P<0.01）;除归脾汤组外，与正常对照组比较，各组NOS3检测值显示明显下调（P<0.05）;与正常对照组比较，实验各组PGI2检测值明显下调（P>0.01）;与正常对照组比较，实验各组TM检测值明显下调（P>0.05）。结论：从脾论治方升高ITP模型小鼠外周血PLT计数以及有效止血与上调血管促凝因子TXA2、VCAM-1，下调PGI2、TM检测值，平衡促凝与抗凝因子密切相关。

关键词 脾主统血;脾不统血证;从脾论治;健脾益气摄血方;免疫性血小板减少症;血管活性因子;血管促凝因子;血管抗凝因子

Hemostatic Mechanism of “Treated From Spleen” Therapy on ITP Based on the Characteristics of Vasoactive Factors

WANG Jun1，ZHANG Yayue2，WANG Jia3，WANG Chong2，CHEN Ke4，CHEN Xinyi2，LANG Haiyan5

（1 The First Affiliated Hospital of Zhejiang Chinese Medical University，Hangzhou 310006，China; 2 Dongzhimen Hospital，Beijing University of Chinese Medicine，Beijing 100700，China; 3 Henan Cancer Hospital，Zhengzhou 450008，China; 4 Shunyi Hospital，Beijing Traditional Chinese Medicine Hospital，Beijing 101300，China; 5 Dongfang Hospital，Beijing University of Chinese Medicine，Beijing 100078，China）

Abstract Objective：To explore the ITP hemostatic mechanism of “treating from spleen” therapy based on the characteristics of vasoactive factors.Methods：The ITP mouse model was established by passive immunomodeling method.The prednisone，Jianpi Yiqi Formula，Jianpi Yiqi Shexue Formula and Guipi Decoction were used as intervention drugs.The content of serum ET-1，NO，NOS3，TXA2，PGI2，VWF，VCAM-1 and TM of the model mice were detected by enzyme-linked immunosorbent assay （ELISA）.Results：1）Therapeutic effect of platelet enhancement：compared with normal control group，PLT count in peripheral blood of mice in each group was significantly decreased after APS injection （P<0.01）; compared with model control group，PLT count in peripheral blood of mice in each group was significantly increased on the 8th day of injection （P<0.01）.2）Hemostasis effect：except for the normal control group，all experimental groups had bleeding tendency to a greater or lesser extent before injection; compared with model control group，the grade of bleeding in experimental groups decreased on the 6th and 8th day of administration （P<0.01）.3）Vascular coagulation factor：compared with normal group，ET-1 detection value in each group was significantly decreased （P<0.01）; compared with normal control group and model control group，TXA2 detection values in experimental groups were significantly increased （P<0.01）.Compared with normal group，VWF detection values in experimental groups were significantly down-regulated （P<0.01）.Compared with normal control group，VCAM-1 detection value in experimental groups was significantly reduced （P<0.01）.4）Vascular anticoagulant factor：compared with normal group，NO detection value in experimental groups was significantly decreased （P<0.01）; Compared with normal control group，NOS3 values in all groups were expressively reduced （P<0.05） except for Guipi Decoction group.Compared with normal control group，PGI2 detection value in experimental groups was significantly cut down （P<0.01）.Compared with normal control group，TM detection values in experimental groups were greatly reduced （P>0.05）.Conclusion：PLT count in peripheral blood of ITP model mice from the treatment of spleen theory is increased，and effectively hemostasis is taken，which has a close correlation between increasing TXA2 and VCAM-1，PGI2 and TM detection values are decreased，and balancing pro-coagulation and anticoagulation factors.

Keywords Spleen governing blood; Splenic failure in controlling blood; Treating from spleen therapy; Jianpi Yiqi Shexue Decoction; Immune thrombocytopenia; Vasoactive factor; Vascular coagulation factor; Vascular anticoagulant factor

中图分类号：R242;R558+.2文献标识码：Adoi：10.3969/j.issn.1673-7202.2021.03.003

免疫性血小板减少症（Immune Thrombocytopenia，ITP）是临床常见的血液系统出血性疾病，主要临床表现为皮肤、黏膜、牙龈、鼻黏膜出血，尿血、便血或月经过多，或月经期长。严重者可有内脏出血。目前，对ITP发病机制尚不完全清晰，公认的发病机制是多因素导致的自身免疫功能紊乱[1-2]。按照中医理论，结合慢性ITP临床特征，其发病是由于脾气虚弱，不能统摄血液而致[3]。因此，“从脾论治”是ITP的基本原则[4-5]。我们在前期临床与实验研究基础上，采用被动免疫造模法复制了ITP动物模型[6-8]，以模型小鼠外周血血小板（Platelet，PLT）计数、出血程度分级与血清内皮素-1（Endothelin-1，ET-1）、血栓素A2（Thromboxane A2，TXA2）、一氧化氮（Nitric Oxide，NO）、一氧化氮合酶NOS3（Nitric Oxide Synthase 3，NOS3）、前列环素（Prostacyclin，PGI2）、血栓调节蛋白（Thrombomodulin，TM）、血管性血友病因子（von Willebrand Factor，vWF）、血管内皮细胞血管细胞黏附分子1（Vascular Cell Adhesion Molecule1，VCAM1）为检测指标。试……