中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2013年
1期
23-27
,共5页
陈楠%刘应莉%刘文天%王邦茂%孙朝侠%王玫%韩明哲
陳楠%劉應莉%劉文天%王邦茂%孫朝俠%王玫%韓明哲
진남%류응리%류문천%왕방무%손조협%왕매%한명철
骨髓移植%间充质干细胞移植%肝炎,自身免疫性%T淋巴细胞%小鼠,近交C57BL
骨髓移植%間充質榦細胞移植%肝炎,自身免疫性%T淋巴細胞%小鼠,近交C57BL
골수이식%간충질간세포이식%간염,자신면역성%T림파세포%소서,근교C57BL
Bone marrow transplantation%Mesenchymal stem cell transplantation%Hepatitis,autoimmune%T-lymphocytes%Mice,inbred C57BL
目的 探讨骨髓间充质干细胞(MSC)移植对小鼠自身免疫性肝炎(AIH)的治疗作用及机制.方法 采用C57BL/6小鼠肝特异性抗原S-100与弗氏完全佐剂诱导同种系44只小鼠建立AIH模型.造模后取6只小鼠验证AIH模型,余38只分为3组,MSC移植组(A组)14只进行MSC尾静脉注射治疗,地塞米松(DXM)治疗组(B组)12只予DXM腹腔注射治疗,PBS对照组(C组)12只予腹腔注射PBS.健康对照组(D组)18只未造模未予任何处理.于实验第5、9周测小鼠体质量,测血清ALT水平,处死各组小鼠取肝组织进行病理学检查并行肝脏炎症活动度Knodel1评分,分离小鼠脾脏T淋巴细胞行增殖抑制实验.统计学分析采用秩和检验、方差分析和t检验.结果 A、B组小鼠治疗后体质量均呈上升趋势(F=15.678,P＜0.01;F=3.730,P=0.037),C组治疗前后变化差异无统计学意义(P＞0.05).实验第5、9周,A、B组小鼠血清ALT水平逐渐下降,各时间点比较差异有统计学意义(F=20.267,P＜0.01；F=4.277,P=0.034)；C组小鼠血清ALT水平治疗前后变化差异无统计学意义(P＞0.05).实验第5、9周,A组小鼠血清ALT降低程度大于B组,差异均有统计学意义(t=3.566、3.218,P均＜0.05).实验第5、9周,A、B组小鼠Knodell评分逐渐下降,各时间点比较差异有统计学意义(F=8.070,P=0.003; F=6.547,P=0.009)；C组小鼠Knodell评分治疗前后变化差异无统计学意义(P＞0.05).实验第9周时,A、B、C三组小鼠Knodell评分间差异有统计学意义(F=4.477,P=0.029).体外脾淋巴细胞增殖抑制实验显示,MSC培养上清液能显著抑制S-100抗原和刀豆蛋白A刺激的T淋巴细胞增殖,吸光度值分别为0.267±0.167比0.217±0.128和0.165±0.187比0.082±0.051,差异均有统计学意义(t=7.187、4.602,P均＜0.01).结论 MSC移植可能通过抑制T淋巴细胞的增殖对小鼠AIH发挥治疗作用.
目的 探討骨髓間充質榦細胞(MSC)移植對小鼠自身免疫性肝炎(AIH)的治療作用及機製.方法 採用C57BL/6小鼠肝特異性抗原S-100與弗氏完全佐劑誘導同種繫44隻小鼠建立AIH模型.造模後取6隻小鼠驗證AIH模型,餘38隻分為3組,MSC移植組(A組)14隻進行MSC尾靜脈註射治療,地塞米鬆(DXM)治療組(B組)12隻予DXM腹腔註射治療,PBS對照組(C組)12隻予腹腔註射PBS.健康對照組(D組)18隻未造模未予任何處理.于實驗第5、9週測小鼠體質量,測血清ALT水平,處死各組小鼠取肝組織進行病理學檢查併行肝髒炎癥活動度Knodel1評分,分離小鼠脾髒T淋巴細胞行增殖抑製實驗.統計學分析採用秩和檢驗、方差分析和t檢驗.結果 A、B組小鼠治療後體質量均呈上升趨勢(F=15.678,P＜0.01;F=3.730,P=0.037),C組治療前後變化差異無統計學意義(P＞0.05).實驗第5、9週,A、B組小鼠血清ALT水平逐漸下降,各時間點比較差異有統計學意義(F=20.267,P＜0.01；F=4.277,P=0.034)；C組小鼠血清ALT水平治療前後變化差異無統計學意義(P＞0.05).實驗第5、9週,A組小鼠血清ALT降低程度大于B組,差異均有統計學意義(t=3.566、3.218,P均＜0.05).實驗第5、9週,A、B組小鼠Knodell評分逐漸下降,各時間點比較差異有統計學意義(F=8.070,P=0.003; F=6.547,P=0.009)；C組小鼠Knodell評分治療前後變化差異無統計學意義(P＞0.05).實驗第9週時,A、B、C三組小鼠Knodell評分間差異有統計學意義(F=4.477,P=0.029).體外脾淋巴細胞增殖抑製實驗顯示,MSC培養上清液能顯著抑製S-100抗原和刀豆蛋白A刺激的T淋巴細胞增殖,吸光度值分彆為0.267±0.167比0.217±0.128和0.165±0.187比0.082±0.051,差異均有統計學意義(t=7.187、4.602,P均＜0.01).結論 MSC移植可能通過抑製T淋巴細胞的增殖對小鼠AIH髮揮治療作用.
목적 탐토골수간충질간세포(MSC)이식대소서자신면역성간염(AIH)적치료작용급궤제.방법 채용C57BL/6소서간특이성항원S-100여불씨완전좌제유도동충계44지소서건립AIH모형.조모후취6지소서험증AIH모형,여38지분위3조,MSC이식조(A조)14지진행MSC미정맥주사치료,지새미송(DXM)치료조(B조)12지여DXM복강주사치료,PBS대조조(C조)12지여복강주사PBS.건강대조조(D조)18지미조모미여임하처리.우실험제5、9주측소서체질량,측혈청ALT수평,처사각조소서취간조직진행병이학검사병행간장염증활동도Knodel1평분,분리소서비장T림파세포행증식억제실험.통계학분석채용질화검험、방차분석화t검험.결과 A、B조소서치료후체질량균정상승추세(F=15.678,P＜0.01;F=3.730,P=0.037),C조치료전후변화차이무통계학의의(P＞0.05).실험제5、9주,A、B조소서혈청ALT수평축점하강,각시간점비교차이유통계학의의(F=20.267,P＜0.01；F=4.277,P=0.034)；C조소서혈청ALT수평치료전후변화차이무통계학의의(P＞0.05).실험제5、9주,A조소서혈청ALT강저정도대우B조,차이균유통계학의의(t=3.566、3.218,P균＜0.05).실험제5、9주,A、B조소서Knodell평분축점하강,각시간점비교차이유통계학의의(F=8.070,P=0.003; F=6.547,P=0.009)；C조소서Knodell평분치료전후변화차이무통계학의의(P＞0.05).실험제9주시,A、B、C삼조소서Knodell평분간차이유통계학의의(F=4.477,P=0.029).체외비림파세포증식억제실험현시,MSC배양상청액능현저억제S-100항원화도두단백A자격적T림파세포증식,흡광도치분별위0.267±0.167비0.217±0.128화0.165±0.187비0.082±0.051,차이균유통계학의의(t=7.187、4.602,P균＜0.01).결론 MSC이식가능통과억제T림파세포적증식대소서AIH발휘치료작용.
Objective To explore the therapeutic effects and mechanism of bone marrow mesenchymal stem cells (MSC) transplantation in mice autoimmune hepatitis (AIH).Methods AIH model was established in 44 C57BL/6 mice,which were induced by homologous series liver-specific antigen S-100 and Freund's complete adjuvant.After modeling,six mice were collected for AIH model confirming.The other 38 mice were divided into three groups.Fourteen mice of MSC transplantation group (group A) were treated by MSC tail vein injection,12 mice of dexamethasone (DXM) group (group B) were treated by DXM intraperitoneal injection,and 12 mice of PBS control group (group C) received phosphate buffer saline (PBS) intraperitoneal injection.Eighteen mice of healthy control group (group D) weren't modeled and received no treatment.At the 5th and 9th week,the mice weights and serum alanine aminotransferase (ALT) level were tested,mice liver tissues of each group were estimated by pathological examination and Knodell scoring,and spleen T lymphocytes of mice were isolated for proliferation-inhibition examination.The data were analyzed by rank sum test,ANOVA and t test.Results After treatment,mice weights of both group A and B showed upward trend (F=15.678,P＜0.01; F=3.730,P=0.037).Before and after treatment,there was no significant difference in group C (P＞0.05).At the 5th and 9th week,the ALT level of group A and B gradually decreased,there was statistical significance between the time points (F=20.267,P＜0.01; F=4.277,P=0.034).Before and after treatment,there was no significant difference in ALT level of group C (P＞0.05).At the 5th and 9th week,the degree of mice serum ALT reduction of group A was larger than that of group B,and the difference was statistically significant (t=3.566 and 3.218,both P＜0.05).At the 5th and 9th week,the Kondell scores of group A and B gradually decreased,there was statistical significance between the time points (F=8.070,P=0.003; F=6.547,P=0.009).Before and after treatment,there was no significant difference in Kondell scores of group C (P＞0.05).At the 9th week,there was statistical significance in Kondell scores among group A,group B and group C (F =4.477,P =0.029).The in vitro spleen lymphocytes proliferation-inhibition experiment demonstrated that the supernatant of MSC could significantly inhibit the proliferation of T lymphocytes stimulated by S-100 antigen and concanavalin A,the absorbance values were0.267±0.167 vs.0.217±0.128 and0.165±0.187 vs.0.082±0.051 respectively,the differences were statistically significant (t =7.187 and 4.602,both P＜ 0.01).Conclusion MSC transplantation may play a therapeutic role in mice AIH through inhibiting T lymphocyte proliferation.
