中华器官移植杂志
中華器官移植雜誌
중화기관이식잡지
CHINESE JOURNAL OF ORGAN TRANSPLANTATION
2013年
4期
244-247
,共4页
崔瀚文%袁清%洪善娟%韦星%许亮%黄海燕%王全兴%蔡明
崔瀚文%袁清%洪善娟%韋星%許亮%黃海燕%王全興%蔡明
최한문%원청%홍선연%위성%허량%황해연%왕전흥%채명
树突状细胞%肾脏%乌司他丁%成熟%趋化能力
樹突狀細胞%腎髒%烏司他丁%成熟%趨化能力
수돌상세포%신장%오사타정%성숙%추화능력
Dendritic cell%Kidney%Ulinastatin%Maturation%Chemotaxis
目的 研究乌司他丁对脂多糖介导的小鼠肾脏固有树突状细胞(rDC)成熟的影响.方法 取C57BL/6J小鼠肾脏,制备单细胞悬液,使用免疫磁珠法分选CD11c+的rDC,流式细胞仪鉴定rDC纯度.脂多糖刺激24 h,促使rDC成熟,给予不同浓度(50、100和200 U/L)乌司他丁处理,酶联免疫吸附试验检测上清液中白细胞介素12(IL-12)的分泌水平.100 U/L的乌司他丁处理后,流式细胞仪检测rDC表面主要组织相容性复合物(MHC)Ⅱ类分子及CD80和CD86分子,以观察乌司他丁对rDC表型及功能的影响.检测趋化因子受体7(CCR7)的表达,计数受巨噬细胞炎症蛋白3β(MIP-3β)趋化进入细胞培养小室下层的CD11c+细胞的数目,分析乌司他丁对rDC趋化能力的影响.结果脂多糖刺激促使rDC成熟,高表达MHCⅡ、CD80、CD86分子,并分泌大量IL-12,而乌司他丁能够抑制脂多糖介导的rDC表型和功能成熟.经乌司他丁处理,CCR7分子的表达由69.2％下降至37％,并且受趋化的CD11c+细胞数目下降为脂多糖处理者的45.3％.结论 乌司他丁能够抑制脂多糖介导的小鼠rDC的成熟和趋化能力上调.
目的 研究烏司他丁對脂多糖介導的小鼠腎髒固有樹突狀細胞(rDC)成熟的影響.方法 取C57BL/6J小鼠腎髒,製備單細胞懸液,使用免疫磁珠法分選CD11c+的rDC,流式細胞儀鑒定rDC純度.脂多糖刺激24 h,促使rDC成熟,給予不同濃度(50、100和200 U/L)烏司他丁處理,酶聯免疫吸附試驗檢測上清液中白細胞介素12(IL-12)的分泌水平.100 U/L的烏司他丁處理後,流式細胞儀檢測rDC錶麵主要組織相容性複閤物(MHC)Ⅱ類分子及CD80和CD86分子,以觀察烏司他丁對rDC錶型及功能的影響.檢測趨化因子受體7(CCR7)的錶達,計數受巨噬細胞炎癥蛋白3β(MIP-3β)趨化進入細胞培養小室下層的CD11c+細胞的數目,分析烏司他丁對rDC趨化能力的影響.結果脂多糖刺激促使rDC成熟,高錶達MHCⅡ、CD80、CD86分子,併分泌大量IL-12,而烏司他丁能夠抑製脂多糖介導的rDC錶型和功能成熟.經烏司他丁處理,CCR7分子的錶達由69.2％下降至37％,併且受趨化的CD11c+細胞數目下降為脂多糖處理者的45.3％.結論 烏司他丁能夠抑製脂多糖介導的小鼠rDC的成熟和趨化能力上調.
목적 연구오사타정대지다당개도적소서신장고유수돌상세포(rDC)성숙적영향.방법 취C57BL/6J소서신장,제비단세포현액,사용면역자주법분선CD11c+적rDC,류식세포의감정rDC순도.지다당자격24 h,촉사rDC성숙,급여불동농도(50、100화200 U/L)오사타정처리,매련면역흡부시험검측상청액중백세포개소12(IL-12)적분비수평.100 U/L적오사타정처리후,류식세포의검측rDC표면주요조직상용성복합물(MHC)Ⅱ류분자급CD80화CD86분자,이관찰오사타정대rDC표형급공능적영향.검측추화인자수체7(CCR7)적표체,계수수거서세포염증단백3β(MIP-3β)추화진입세포배양소실하층적CD11c+세포적수목,분석오사타정대rDC추화능력적영향.결과지다당자격촉사rDC성숙,고표체MHCⅡ、CD80、CD86분자,병분비대량IL-12,이오사타정능구억제지다당개도적rDC표형화공능성숙.경오사타정처리,CCR7분자적표체유69.2％하강지37％,병차수추화적CD11c+세포수목하강위지다당처리자적45.3％.결론 오사타정능구억제지다당개도적소서rDC적성숙화추화능력상조.
Objective To investigate the inhibitory effect of ulinastatin (UTI),an ulinary trypsin inhibitor,on the maturation of renal dendritic cells (rDCs) induced by LPS stimulation.Method The kidneys from C57BL/6J (H-2Kb) mice were harvested to make single cell suspension.rDCs were then selected by Magnetic Cell Sorting (MACS) and assessed for content and purity by flow cytometry.The matured rDCs were generated by stimulating rDCs with LPS for 24 h,then treated with different concentrations of UTI (0-200 U/L).LPS-stimulated cell supernatants were collected for IL-12 measurement by using ELISA.We used 100U/L of UTI to treat matured rDCs,and assessed the maturation phenotype as well as chemokine receptor expression of DCs by using flow cytometry.For chemotaxis assay,LPS-stimulated rDCs treated with UTI (100U/L) were loaded into upper chemotaxis chambers.Cells migrating into the lower chambers were counted by using flow cytometry.Result LPS-stimulated rDCs secreted high levels of IL-12 and expressed high levels of CD80,CD86 and MHC class Ⅱ,and UTI treatment significantly inhibited the production of IL-12 and expression of CD80,CD86 and MHC class Ⅱ.UTI treatment further suppressed the expression of CCR7 on rDCs,as well as their migratory responses to MIP-3β upon LPS stimulation.Conclusion UTI treatment inhibits LPS-mediated maturation of rDCs and interferes with the up-regulation of rDCs chemotaxis.
