摘要
目的:建立肠淋巴液外泌体分离技术,验证外泌体来源,观察星状神经节阻滞(SGB)对失血性休克后肠淋巴液(PHSML)中外泌体数量的影响。方法:雄性大鼠随机均分为假手术(sham)组、sham+SGB组、休克(shock)组和shock+SGB组,常规方法实施SGB并建立失血性休克模型,引流PHSML,提取外泌体。应用纳米粒径分析、CD63蛋白表达检测鉴定提取物是否为外泌体;检测外泌体肠上皮细胞黏附分子(EpCAM)的蛋白表达,明确外泌体是否源于肠上皮细胞。应用流式细胞术分析外泌体数量。结果:肠淋巴液提取的颗粒样物质直径大小在100 nm左右,同时表达外泌体的特异性蛋白CD63和肠道上皮细胞的特异性蛋白EpCAM;shock组肠淋巴液外泌体数量显著高于sham组(P<0.05),shock+SGB组肠淋巴液外泌体数量显著低于shock组(P<0.05)。结论:我们建立了肠淋巴液外泌体的提取技术,证实肠淋巴液外泌体来自肠道上皮细胞,SGB可以降低大鼠失血性休克后肠淋巴液中外泌体的数量。
AIM:To isolate the exosomes in mesenteric lymph,verify the source of exosomes,and observe the effect of stellate ganglion block(SGB)on the number of exosomes in post-hemorrhagic shock mesenteric lymph(PHSML)of rats.METHODS:Twenty-four male rats were randomly divided into sham,sham+SGB,shock,and shock+SGB groups.SGB was performed before the establishment of hemorrhagic shock model using the routine methods in our lab.The PHSML was drained for exosomes isolation.The exosomes were identified through particle size analysis and CD63 protein expression.The expression of epithelial cell adhesion molecule(EpCAM)was detected to identify whether the exosomes were derived from epithelial cell.The number of exosomes in various mesenteric lymphs was measured using the flow cytometry.RESULTS:The diameter of granular material extracted from mesenteric lymph was about 100 nm.The positive expression of exosomes pecific protein CD63 indicated the successful isolation of exosomes,and the EpCAM expression verified the exosomes were derived from intestinal epithelial cells.The number of exosomes in mesenteric lymph isolated from the rats of Shock group was obviously increased compared to that from the Sham group(P<0.05),while the exosomes from the Shock+SGB group was markedly decreased when compared to Shock group(P<0.05).CON⁃CLUSION:The current study establishes the isolation technique of exosomes in mesenteric lymph,and proved the exo⁃somes were derived from the intestinal epithelial cells.SGB treatment reduces the number of exosomes in PHSML.
作者
范泽华
郜思齐
杜会博
张立民
张红
赵振奥
赵自刚
牛春雨
FAN Ze-hua;GAO Si-qi;DU Hui-bo;ZHANG Li-min;ZHANG Hong;ZHAO Zhen-ao;ZHAO Zi-gang;NIU Chun-yu(Institute of Microcirculation and Basic Medical College of Hebei North University,Zhangjiakou 075000,China;Basic Medical College of Hebei Medical University,Shijiazhuang 050017,China.)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2020年第11期2074-2080,共7页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81770492)。
