白头翁汤通过保护微血管内皮细胞的完整性及PMNs迁移杀菌功能的影响  

Pulsatillae Decoction Improves the Bactericidal Capacity of Neutrophils by Protection of Integrity of Microvascular Endothelial Cells

作　　者：王明明[1] 杨舒[1] 董虹[1] 穆祥[1] 

机构地区：[1]北京农学院兽医学(中医药)北京市重点实验室,北京102206

出　　处：《畜牧兽医学报》2016年第4期836-843,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然基金项目(31272144;31572558);北京市科委科技计划(Z121100007412004);北京市科技新星(Z141105001814041);北京市自然科学基金(6132007)

摘　　要：本研究旨在研究白头翁汤(PD)对脂多糖(LPS)诱导内皮细胞损伤的保护作用,从而揭示白头翁汤的药理作用。分别采用水溶性四氮唑(WST^(-1))、乳酸脱氢酶(LDH)、苏木素伊红(HE)和Hoechst33342染色法检测PD和LPS对大鼠肠黏膜微血管内皮细胞(RIMVECs)增殖、细胞毒性、细胞连接和细胞核变化的影响;建立中性粒细胞(PMNs)跨RIMVECs迁移的Transwel模型,检测PD对LPS损伤的RIMVECs后PMNs细胞内和细胞外细菌数量的影响。结果显示,LPS(1μg·mL^(-1))致RIMVECs细胞核萎缩、细胞膜损坏,细胞毒性百分比达53.2%;细胞连接破坏导致细胞脱落,脱落率为39.6%。PD(20μg·mL^(-1))对上述损伤有明显的保护作用。在20μg·mL^(-1)PD作用下,LPS诱导的跨内皮迁移的PMNs细胞内外细菌的存活明显减少(P<0.01)。结果揭示PD可以有效保护LPS损伤的RIMVECs从而增强PMNs的杀菌能力,为临床治疗动物细菌性疾病用药提供理论依据。The aims of this paper was to investigate the protective effect of pulsatillae decoction（PD）on endothelial cells injured by lipopolysaccharide（LPS）and to reveal the pharmacological of PD.The cell proliferation and cytotoxicity of the rat microvascular endothelial cells（RIMVECs）were determined by WST^（-1）ELISA kit and lactate dehydrogenase（LDH）assay.In addition,the RIMVECs damage stimulated by LPS was observed by hematoxylin-eosin（HE）and Hoechst 33342 staining.The transedothlial neutrophils killing was detected by gentamicin protection assay though the build of Transwell.The results showed that LPS（1μg·mL^（-1））make the nuclei of RIMVECs shriveled and damage the cell membrane of RIMVECs and lead to cell fall off.The rate of cytotoxicity was 53.2%.The rate of separated cells wass 39.6%.PD（20μg·mL^（-1））had the significant proliferative effect for that.LPS induced transendothelial migration of neutrophils can reduce the survival of bacteria in both intracellular and extracellular under the effect of 20μg·mL^（-1） PD（P〈0.01）.Results reveal that PD can improve the sterilization function of PMNsby the effective protection of LPS-induced injury of RIMVECs,and might improve the sterilization function of PMNs by protecting the integrity of the RIMVECs to provide a theoretical basis for the dose of clinical treatment of bacterial animal diseases.

关 键 词：白头翁汤 大鼠肠黏膜微血管内皮细胞 脂多糖 中性粒细胞 

分 类 号：S853.9[农业科学—临床兽医学]