中华小儿外科杂志
中華小兒外科雜誌
중화소인외과잡지
CHINESE JOURNAL OF PEDIATRIC SURGERY
2013年
1期
55-59
,共5页
胆道闭锁%流式细胞术%小鼠
膽道閉鎖%流式細胞術%小鼠
담도폐쇄%류식세포술%소서
Biliary atresia%Flow cytometry%Mice
目的 研究轮状病毒感染致新生鼠胆道闭锁不同时段肝内外各亚型炎症细胞浸润状态,并初步探讨调控新生鼠胆道闭锁肝内外炎症反应的主要炎症类型.方法 选取健康新生Balb/C小鼠,出生24 h内使用轮状病毒腹腔注射为实验组,注射等量的轮状病毒培养液(DMEM)为对照组.注射病毒后第7d、14d分别处死新生鼠并获取胆管、肝脏及脾脏组织标本.根据新生鼠皮肤黄染、体重变化将实验组分为致胆道闭锁组(BA+)与未致胆道闭锁组(BA-),再根据胆管HE染色结果,验证分组的准确性.利用流式细胞技术,分别检测各组在不同时段肝脏及脾脏组织内CD4+ Th、CD8+Tc、NK细胞、巨噬细胞(Mac)及T淋巴调节细胞(Treg)的浸润状态,并进行统计学分析.结果 脾脏组织:三组间炎症细胞浸润的类型及数量均无明显统计学差异(P>0.05);肝脏组织:轮状病毒感染7 d后BA-组CD4+ Th、Mac及BA+组CD4+ Th、CD8+ Tc、NK、Mac、Treg与对照组相比表达明显升高(P<0.05),BA+组CD8+ Te、NK、Treg表达又明显高于BA-组(P<0.05);感染14d后BA-组CD4+ Th及BA+组CD4+ Th、CD8+ Tc、NK、Treg与对照组相比表达显著升高(P<0.05),BA+组CD8+ Tc、NK、Treg表达亦明显高于BA组(P<0.05);CD4+ Th在BA-组及BA+组均表现为持续升高;CD8+ Te、Treg仅在BA+组表现为持续升高,而Mac表现为先升高后降低.结论 胆道闭锁新生鼠肝内存在特异性渐进性炎症反应,CD4+T淋巴辅助细胞、CD8+T淋巴杀伤细胞、NK细胞、巨噬细胞及T淋巴调节细胞广泛激活并可能共同参与了这一特异性炎症反应.
目的 研究輪狀病毒感染緻新生鼠膽道閉鎖不同時段肝內外各亞型炎癥細胞浸潤狀態,併初步探討調控新生鼠膽道閉鎖肝內外炎癥反應的主要炎癥類型.方法 選取健康新生Balb/C小鼠,齣生24 h內使用輪狀病毒腹腔註射為實驗組,註射等量的輪狀病毒培養液(DMEM)為對照組.註射病毒後第7d、14d分彆處死新生鼠併穫取膽管、肝髒及脾髒組織標本.根據新生鼠皮膚黃染、體重變化將實驗組分為緻膽道閉鎖組(BA+)與未緻膽道閉鎖組(BA-),再根據膽管HE染色結果,驗證分組的準確性.利用流式細胞技術,分彆檢測各組在不同時段肝髒及脾髒組織內CD4+ Th、CD8+Tc、NK細胞、巨噬細胞(Mac)及T淋巴調節細胞(Treg)的浸潤狀態,併進行統計學分析.結果 脾髒組織:三組間炎癥細胞浸潤的類型及數量均無明顯統計學差異(P>0.05);肝髒組織:輪狀病毒感染7 d後BA-組CD4+ Th、Mac及BA+組CD4+ Th、CD8+ Tc、NK、Mac、Treg與對照組相比錶達明顯升高(P<0.05),BA+組CD8+ Te、NK、Treg錶達又明顯高于BA-組(P<0.05);感染14d後BA-組CD4+ Th及BA+組CD4+ Th、CD8+ Tc、NK、Treg與對照組相比錶達顯著升高(P<0.05),BA+組CD8+ Tc、NK、Treg錶達亦明顯高于BA組(P<0.05);CD4+ Th在BA-組及BA+組均錶現為持續升高;CD8+ Te、Treg僅在BA+組錶現為持續升高,而Mac錶現為先升高後降低.結論 膽道閉鎖新生鼠肝內存在特異性漸進性炎癥反應,CD4+T淋巴輔助細胞、CD8+T淋巴殺傷細胞、NK細胞、巨噬細胞及T淋巴調節細胞廣汎激活併可能共同參與瞭這一特異性炎癥反應.
목적 연구륜상병독감염치신생서담도폐쇄불동시단간내외각아형염증세포침윤상태,병초보탐토조공신생서담도폐쇄간내외염증반응적주요염증류형.방법 선취건강신생Balb/C소서,출생24 h내사용륜상병독복강주사위실험조,주사등량적륜상병독배양액(DMEM)위대조조.주사병독후제7d、14d분별처사신생서병획취담관、간장급비장조직표본.근거신생서피부황염、체중변화장실험조분위치담도폐쇄조(BA+)여미치담도폐쇄조(BA-),재근거담관HE염색결과,험증분조적준학성.이용류식세포기술,분별검측각조재불동시단간장급비장조직내CD4+ Th、CD8+Tc、NK세포、거서세포(Mac)급T림파조절세포(Treg)적침윤상태,병진행통계학분석.결과 비장조직:삼조간염증세포침윤적류형급수량균무명현통계학차이(P>0.05);간장조직:륜상병독감염7 d후BA-조CD4+ Th、Mac급BA+조CD4+ Th、CD8+ Tc、NK、Mac、Treg여대조조상비표체명현승고(P<0.05),BA+조CD8+ Te、NK、Treg표체우명현고우BA-조(P<0.05);감염14d후BA-조CD4+ Th급BA+조CD4+ Th、CD8+ Tc、NK、Treg여대조조상비표체현저승고(P<0.05),BA+조CD8+ Tc、NK、Treg표체역명현고우BA조(P<0.05);CD4+ Th재BA-조급BA+조균표현위지속승고;CD8+ Te、Treg부재BA+조표현위지속승고,이Mac표현위선승고후강저.결론 담도폐쇄신생서간내존재특이성점진성염증반응,CD4+T림파보조세포、CD8+T림파살상세포、NK세포、거서세포급T림파조절세포엄범격활병가능공동삼여료저일특이성염증반응.
Objective To explore the inflammatory cellular profiles in the liver and spleen in the rotavirus (RRV) mouse model of biliary atresia (BA).Methods Balb/c neoratal mice were used in this experiment.Within the first 24 hours after birth,mice were intraperitoneally injected with RRV to induce BA.The control mice were injected with RRV culture medium DMEM.The mice were sacrificed 7 or 14 days after RRV infection.Extrahepatic bile duct,liver and spleen were harvested for further study.The RRV infected mice were divided into the biliary atresia group (BA) and non-biliary atresia group (nBA).The diagnosis of BA was confirmed by obliteration of bile duct on H&E staining.The inflammatory cellular profiles including CD4+ T lymphocytes helper cells (CD4+ Th),CD8+cytotoxic T lymphocytes (CD8+ Tc),NK cells and Macrophages (Mac) and regulatory T cells (Treg)were determined by flow cytometry.Results In the spleen,there was no significant differences of CD4+ Th,CI38+ Tc,Mac and Treg between the BA mice and control mice either 7 or 14 days after RRV infection (all P>0.05).In the liver,7 days after RRV infection,CID+ Th,CD8+ Tc,NK,Mac and Treg in BA mice were significantly increased than those of control mice and the nBA mice (all,P<0.05).Mac of the nBA mice was increased significantly than that of control mice (P<0.05).In the liver,14 days after RRV infection,CD4 + Th,CD8 + Tc,NK,Mac and Treg in BA mice were significantly increased than those of control mice (all P<0.05).C1D8 + Tc,NK,Mac and Treg in BA mice were significantly increased than those of control mice and the nBA mice (all P<0.05).CD4 + Th of the nBA mice was also increased significantly than that of control mice (P<0.05).At the day 14 after infection,CD8 + Tc,NK,and Treg in BA group were significantly higher than those of control group and nBA group (P<0.05).CD4 + Th in nBA group and BA group was significantly higher than that of control group (P<0.05).In RRV infected mice,CD4 + Th was consistently increased until after 14 days.However,CD8 + Tc and Treg were increased until 14 days only in BA mice.Mac peaked on the 7 day after RRV infection,and then decreased.Conclusions The inflammatory cellular profile in the live is dynamically changed in the course of BA development,which may be associated with the pathogenesis of biliary atresia.