[1]孙祯,黄赤兵,宋亚军,等.大鼠肠系膜淋巴结和外周淋巴结固有exosome的制备及其对T细胞增殖的影响[J].第三军医大学学报,2013,35(04):307-310.
 Sun Zhen,Huang Chibing,Song Yajun,et al.Purification of exosomes from rat mesenteric and peripheral lymph nodes and their effect on T cell proliferation[J].J Third Mil Med Univ,2013,35(04):307-310.
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大鼠肠系膜淋巴结和外周淋巴结固有exosome的制备及其对T细胞增殖的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
35卷
期数:
2013年第04期
页码:
307-310
栏目:
论著
出版日期:
2013-02-28

文章信息/Info

Title:
Purification of exosomes from rat mesenteric and peripheral lymph nodes and their effect on T cell proliferation
作者:
孙祯黄赤兵宋亚军余雪松贺伟峰陈益荣胡文刚王晋
第三军医大学新桥医院泌尿外科第三军医大学西南医院全军烧伤研究所,创伤、烧伤与复合伤国家重点实验室
Author(s):
Sun Zhen Huang Chibing Song Yajun Yu Xuesong He Weifeng Chen Yirong Hu Wengang Wang Jin
Department of Urology, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037; State Key Laboratory of Trauma, Burns and Combined Injuries, Institute of Burns, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China
关键词:
exosome淋巴结超滤密度梯度离心混合淋巴细胞培养
Keywords:
exosome lymphoid nodes ultrafiltration and sucrose gradient centrifugation mixed lymphocyte culture
分类号:
R322.25;R392-33;R392.12
文献标志码:
A
摘要:
目的      从大鼠肠系膜淋巴结、外周淋巴结中分离纯化exosome,鉴定其对T细胞增殖的影响。      方法      取出BN(RT1n)大鼠的外周淋巴结及肠系膜淋巴结并研磨、离心以获得非细胞组织上清,采用超滤密度梯度离心法提取exosome。透射电镜对收集的exosome进行形态学鉴定,BCA法进行蛋白定量,SDS-PAGE电泳分离蛋白,Western blot检测IL-10 的表达,并通过混合淋巴细胞培养检测其对T细胞增殖的影响。      结果      两种淋巴组织来源的非细胞组织上清均含有exosome,肠系膜淋巴结源性exosome蛋白浓度[(1.92±0.10) μg/μL]显著高于外周淋巴结源性exosome[(1.57±0.06)μg/μL,P<0.05],SDS-PAGE显示两种组织源性exosome所含蛋白种类和数量有明显差异,Western blot显示肠系膜淋巴结固有exosome中IL-10的表达显著高于外周淋巴结固有exosome,混合淋巴细胞培养结果显示肠系膜淋巴结固有exosome和外周淋巴结固有exosome对T细胞的增殖均有促进作用(P<0.05),外周淋巴结源性exosome的促增殖作用显著大于肠系膜淋巴结源性exosome(P<0.05)。      结论      肠系膜淋巴结、外周淋巴结均含有组织固有exosome,两种组织来源exosome有蛋白浓度和蛋白种类的差异,且均能促进淋巴细胞增殖,促进程度有显著差异。
Abstract:
Objective      To investigate the purification methods of the exosomes derived form lymphoid tissues of rats and determine their effect on T cell proliferation.       Methods      The peripheral lymph nodes and mesenteric lymph nodes from BN (RT1n) rats were separated, and their impurity were removed by centrifugation. Then, the exosomes were purified from the acellular supernatants by means of differential ultracentrifugation or ultrafiltration and sucrose gradient centrifugation, and confirmed by transmission electron microscopy. The protein expression of the exosomes was analyzed by SDS-PAGE electrophoresis. Western blot analysis was used to test the expression of IL-10. Finally, mixed lymphocyte culture was used to evaluate the immune function of the exosomes from peripheral lymph nodes and mesenteric lymph nodes.       Results      The protein concentration of the exosomes purified from mesenteric lymph nodes were higher than that from peripheral lymph nodes (1.92±0.10 vs 1.57±0.06 μg/μL, P<0.05). SDS-PAGE displayed the differences in protein contents and types between the exosomes from 2 kinds of lymph nodes. Western blotting indicated that the expression of IL-10 was stronger in the exosomes from mesenteric lymph nodes than those from peripheral lymph nodes. The two kinds of exosomes both promoted lymphocyte proliferation (P<0.05), while the exosomes from peripheral lymph nodes exosomes stronger than from mesenteric lymph nodes (P<0.05).       Conclusion      There are exosomes in both peripheral and mesenteric lymph nodes, but with significant differences in protein contents and types. The exosomes from 2 kinds of lymph nodes promote lymphocyte proliferation, though with significant differences.

参考文献/References:

孙祯, 黄赤兵, 宋亚军, 等. 大鼠肠系膜淋巴结和外周淋巴结固有exosome的制备及其对T细胞增殖的影响[J].第三军医大学学报,2013,35(4):307-310.

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更新日期/Last Update: 2013-02-25