中华临床医师杂志（电子版）
中華臨床醫師雜誌（電子版）
중화림상의사잡지（전자판）
CHINESE JOURNAL OF CLINICIANS(ELECTRONIC VERSION)
2013年
24期
11523-11527
,共5页
吴冲%孙华文%曾放%刘顺%吴红学
吳遲%孫華文%曾放%劉順%吳紅學
오충%손화문%증방%류순%오홍학
Tim-3%Crohn病%模型,动物%预测
Tim-3%Crohn病%模型,動物%預測
Tim-3%Crohn병%모형,동물%예측
Tim-3%Crohn disease%Models,animal%Forecasting
目的：探讨T细胞免疫球蛋白及黏蛋白分子-3（Tim-3）在不同病情程度的克罗恩病（CD）小鼠模型建模前后的差异性表达及机制，寻找CD的新型血液标志物。方法80只小鼠采用3种不同浓度的2，4，6-三硝基苯磺酸乙醇溶液经肛门灌肠建立克罗恩小鼠疾病模型，分为4组，高浓度组（n＝10），中浓度组（n＝50），低浓度组（n＝10），对照组（n＝10），中浓度组于给药前、给药第2天，给药第7天，给药第14天，给药第21天5个时间点随机取10只小鼠，其余各浓度组以及对照组于造模后14 d取小鼠外周血行RT-PCR实验测定Tim-3 mRNA的含量，同时ELISA实验测定外周血中IL-4以及INF-γ的含量，取小鼠结肠切片后行HE染色，了解Tim-3 mRNA含量与其他各指标的关联性。结果成功构建了小鼠的CD模型，造模药物浓度越高，小鼠的病理学评分越高，病情越重，各组间差异有统计学意义（P＜0.05）。造模后14 d内，Tim-3 mRNA的表达量逐渐增加，14 d后表达量趋于平稳。平稳期，外周血中Tim-3 mRNA的表达量在高浓度2，4，6-三硝基苯磺酸组（2.12±0.24），中浓度组（1.68±0.18），低浓度组（1.37±0.12）以及正常对照组中（1.00±0.00）的表达依次降低，差异有统计学意义（P＜0.05）。结论外周血中Tim-3的含量在CD小鼠模型起病急性期渐进性增加，给药7 d后趋于稳定，稳定期的Tim-3含量与疾病的严重程度呈正相关，可以作为CD病情程度的预测指标，具有较好的稳定性和敏感性。
目的：探討T細胞免疫毬蛋白及黏蛋白分子-3（Tim-3）在不同病情程度的剋囉恩病（CD）小鼠模型建模前後的差異性錶達及機製，尋找CD的新型血液標誌物。方法80隻小鼠採用3種不同濃度的2，4，6-三硝基苯磺痠乙醇溶液經肛門灌腸建立剋囉恩小鼠疾病模型，分為4組，高濃度組（n＝10），中濃度組（n＝50），低濃度組（n＝10），對照組（n＝10），中濃度組于給藥前、給藥第2天，給藥第7天，給藥第14天，給藥第21天5箇時間點隨機取10隻小鼠，其餘各濃度組以及對照組于造模後14 d取小鼠外週血行RT-PCR實驗測定Tim-3 mRNA的含量，同時ELISA實驗測定外週血中IL-4以及INF-γ的含量，取小鼠結腸切片後行HE染色，瞭解Tim-3 mRNA含量與其他各指標的關聯性。結果成功構建瞭小鼠的CD模型，造模藥物濃度越高，小鼠的病理學評分越高，病情越重，各組間差異有統計學意義（P＜0.05）。造模後14 d內，Tim-3 mRNA的錶達量逐漸增加，14 d後錶達量趨于平穩。平穩期，外週血中Tim-3 mRNA的錶達量在高濃度2，4，6-三硝基苯磺痠組（2.12±0.24），中濃度組（1.68±0.18），低濃度組（1.37±0.12）以及正常對照組中（1.00±0.00）的錶達依次降低，差異有統計學意義（P＜0.05）。結論外週血中Tim-3的含量在CD小鼠模型起病急性期漸進性增加，給藥7 d後趨于穩定，穩定期的Tim-3含量與疾病的嚴重程度呈正相關，可以作為CD病情程度的預測指標，具有較好的穩定性和敏感性。
목적：탐토T세포면역구단백급점단백분자-3（Tim-3）재불동병정정도적극라은병（CD）소서모형건모전후적차이성표체급궤제，심조CD적신형혈액표지물。방법80지소서채용3충불동농도적2，4，6-삼초기분광산을순용액경항문관장건립극라은소서질병모형，분위4조，고농도조（n＝10），중농도조（n＝50），저농도조（n＝10），대조조（n＝10），중농도조우급약전、급약제2천，급약제7천，급약제14천，급약제21천5개시간점수궤취10지소서，기여각농도조이급대조조우조모후14 d취소서외주혈행RT-PCR실험측정Tim-3 mRNA적함량，동시ELISA실험측정외주혈중IL-4이급INF-γ적함량，취소서결장절편후행HE염색，료해Tim-3 mRNA함량여기타각지표적관련성。결과성공구건료소서적CD모형，조모약물농도월고，소서적병이학평분월고，병정월중，각조간차이유통계학의의（P＜0.05）。조모후14 d내，Tim-3 mRNA적표체량축점증가，14 d후표체량추우평은。평은기，외주혈중Tim-3 mRNA적표체량재고농도2，4，6-삼초기분광산조（2.12±0.24），중농도조（1.68±0.18），저농도조（1.37±0.12）이급정상대조조중（1.00±0.00）적표체의차강저，차이유통계학의의（P＜0.05）。결론외주혈중Tim-3적함량재CD소서모형기병급성기점진성증가，급약7 d후추우은정，은정기적Tim-3함량여질병적엄중정도정정상관，가이작위CD병정정도적예측지표，구유교호적은정성화민감성。
Objective Investigate the dysregulated expression of T-cell immunoglobulin and mucin molecules-3 (Tim-3) in mice models with different severity of Crohn's disease, Looking for a new type of Crohn's disease blood marker. Methods the Crohn's disease animal model was made by the anus enema using three different concentrations of 2, 4, 6-trinitrobenzene sulfonic acid ethanol solution, 80 mice were randomly divided into four groups:high concentration group(n=10), moderate concentration group(n=50), low concentration group (n=10), control group(n=10), the expressions of Tim-3 mRNA in peripheral blood were detected by RT-PCR at two days, one week, two weeks, three weeks after we got the mice models in moderate concentration group, we did it two weeks later in the other three groups, At the same time, the content of IL-4 and INF-γin peripheral blood were measured by ELISEA, and colon section from mice underwent HE staining. Results Models of Crohn's disease in mice were successfully constructed, The higher concentration result in higher pathology score as well as more severe disease in mice .the expressions of Tim-3 mRNA in peripheral blood were gradually increased in two weeks after we made the mice models and remained stable after two weeks. At stationary phase, the expressions of Tim-3 mRNA were gradually reduced in high concentration group (2.12 ± 0.24), moderate concentration group(1.68±0.18), low concentration group(1.37±0.12) and Control group (1.00±0.00). And the difference was statistically significant(P<0.05). Conclusion The expression of Tim-3 mRNA in Peripheral blood of a mice model with Crohn's Disease was gradually increased during the first 14 days, and then keep stable, at stationary phase, the expression of Tim-3 mRNA is positively correlated with the severity of the disease. It can be a predictor of disease severity and has good stability and sensitivity.
