中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2009年
9期
932-936
,共5页
脓毒症%调节性T细胞%效应T细胞%凋亡%血必净注射液
膿毒癥%調節性T細胞%效應T細胞%凋亡%血必淨註射液
농독증%조절성T세포%효응T세포%조망%혈필정주사액
Sepsis%Regulatory T cells%Effector T cells%Apoptosis%Xuebijing injection
目的 探讨中药血必净注射液对体外脂多糖(LPS)刺激CD4+CD25+调节性T细胞(Treg)凋亡及其介导效应T细胞(Teff)增殖反应、分泌白细胞介素-2(IL-2)功能的影响.方法 40只雄性清洁级Wistar大鼠,实验前禁食12 h.免疫磁珠法分选大鼠脾脏CD4+CD25+Treg,分为对照组(n=8)、抗CD3/CD28组(n=8)、抗CXB/CD28+LPS组(/1,=8)、抗CD3/CD28+血必净组(n=8)和抗CD3/CD28+LPS+血必净组(n=8).培养3 d后流式细胞术检测CD4+CD25+Treg凋亡率、叉头翼状螺旋转录因子(Foxp3)和T淋巴细胞毒性相关抗原4(CTLA-4)表达,酶联免疫吸附试验检测IL-10分泌.同时,CD4+CD25+Treg与CD4+CD25-T细胞1:1培养,刀豆素A刺激68 h后分析Treg对Teff增殖的抑制效应,并检测上清中IL-2/可溶性IL-2受体水平.结果 抗CD3/CD28诱导Treg凋亡率为(33.70±3.06%),显著高于对照组(12.84±0.84)%;抗CD3/CD28+LPS+血必净组(45.13±2.70)%凋亡率明显高于抗CD3/CD28+LPS组[(29.41±1.63)%,P＜0.01].同时,CD4+CLY25+Treg roxp3和CTLA-4表达及IL-10分泌随凋亡增加而降低.抗CD3/CD28+血必净组(31.26%)Teff增殖反应平均抑制率显著低于对照组(54.48%,P＜0.05),抗CD3/CD28+LPS+血必净组抑制率与抗CD3/CD28+LPS组比较亦明显下降(P＜0.01).此外,抗CD3/CD28+血必净组和抗CD3/CD28+LPS+血必净组IL-2分泌水平均显著高于抗CD3/CD28+LPS组(P＜0.01).结论 LPS刺激可明显上调大鼠Treg对Teff的抑制功能,血必净注射液能通过促进Treg凋亡而有效改善其对Teff的抑制效应.
目的 探討中藥血必淨註射液對體外脂多糖(LPS)刺激CD4+CD25+調節性T細胞(Treg)凋亡及其介導效應T細胞(Teff)增殖反應、分泌白細胞介素-2(IL-2)功能的影響.方法 40隻雄性清潔級Wistar大鼠,實驗前禁食12 h.免疫磁珠法分選大鼠脾髒CD4+CD25+Treg,分為對照組(n=8)、抗CD3/CD28組(n=8)、抗CXB/CD28+LPS組(/1,=8)、抗CD3/CD28+血必淨組(n=8)和抗CD3/CD28+LPS+血必淨組(n=8).培養3 d後流式細胞術檢測CD4+CD25+Treg凋亡率、扠頭翼狀螺鏇轉錄因子(Foxp3)和T淋巴細胞毒性相關抗原4(CTLA-4)錶達,酶聯免疫吸附試驗檢測IL-10分泌.同時,CD4+CD25+Treg與CD4+CD25-T細胞1:1培養,刀豆素A刺激68 h後分析Treg對Teff增殖的抑製效應,併檢測上清中IL-2/可溶性IL-2受體水平.結果 抗CD3/CD28誘導Treg凋亡率為(33.70±3.06%),顯著高于對照組(12.84±0.84)%;抗CD3/CD28+LPS+血必淨組(45.13±2.70)%凋亡率明顯高于抗CD3/CD28+LPS組[(29.41±1.63)%,P＜0.01].同時,CD4+CLY25+Treg roxp3和CTLA-4錶達及IL-10分泌隨凋亡增加而降低.抗CD3/CD28+血必淨組(31.26%)Teff增殖反應平均抑製率顯著低于對照組(54.48%,P＜0.05),抗CD3/CD28+LPS+血必淨組抑製率與抗CD3/CD28+LPS組比較亦明顯下降(P＜0.01).此外,抗CD3/CD28+血必淨組和抗CD3/CD28+LPS+血必淨組IL-2分泌水平均顯著高于抗CD3/CD28+LPS組(P＜0.01).結論 LPS刺激可明顯上調大鼠Treg對Teff的抑製功能,血必淨註射液能通過促進Treg凋亡而有效改善其對Teff的抑製效應.
목적 탐토중약혈필정주사액대체외지다당(LPS)자격CD4+CD25+조절성T세포(Treg)조망급기개도효응T세포(Teff)증식반응、분비백세포개소-2(IL-2)공능적영향.방법 40지웅성청길급Wistar대서,실험전금식12 h.면역자주법분선대서비장CD4+CD25+Treg,분위대조조(n=8)、항CD3/CD28조(n=8)、항CXB/CD28+LPS조(/1,=8)、항CD3/CD28+혈필정조(n=8)화항CD3/CD28+LPS+혈필정조(n=8).배양3 d후류식세포술검측CD4+CD25+Treg조망솔、차두익상라선전록인자(Foxp3)화T림파세포독성상관항원4(CTLA-4)표체,매련면역흡부시험검측IL-10분비.동시,CD4+CD25+Treg여CD4+CD25-T세포1:1배양,도두소A자격68 h후분석Treg대Teff증식적억제효응,병검측상청중IL-2/가용성IL-2수체수평.결과 항CD3/CD28유도Treg조망솔위(33.70±3.06%),현저고우대조조(12.84±0.84)%;항CD3/CD28+LPS+혈필정조(45.13±2.70)%조망솔명현고우항CD3/CD28+LPS조[(29.41±1.63)%,P＜0.01].동시,CD4+CLY25+Treg roxp3화CTLA-4표체급IL-10분비수조망증가이강저.항CD3/CD28+혈필정조(31.26%)Teff증식반응평균억제솔현저저우대조조(54.48%,P＜0.05),항CD3/CD28+LPS+혈필정조억제솔여항CD3/CD28+LPS조비교역명현하강(P＜0.01).차외,항CD3/CD28+혈필정조화항CD3/CD28+LPS+혈필정조IL-2분비수평균현저고우항CD3/CD28+LPS조(P＜0.01).결론 LPS자격가명현상조대서Treg대Teff적억제공능,혈필정주사액능통과촉진Treg조망이유효개선기대Teff적억제효응.
Objective To investigate the effect of Xuebijing injectiong on lipopolysaccharide(LPS)-induced apoptosis of CD4+CD25+regulatory T cells(Tregs)and immune function of effector T cells(Teff)in vitro.Method CD4+CD25+Tregs isolated from rat spleens were divided into the control group,anti-CD3/CD28 group,anti-CD3/CD28+LPS group,anti-CD3/CD28+Xuebijing injection group,and anti-CD3/CD28+LPS+Xuebijing injection group.The apoptosis rate and expression of forkhead/winged helix transcription factor p3 (Foxp3)and cytotoxic T-lymphocyte-associated antigen 4(CTLA-4)of CD4+CD25+Tregs were detected by flow cytometry(FCM),and the secretion of IL-10 of Tregs was measured by ELBA on day 3.CD4+CD25-T cells were co-cultured with CD4+CD25+Tregs(1:1)for 68 hours,proliferative activity of Teff was determined by MTT,and interleukin(IL)-2/sIL-2Rα levels were measured by ELISA.Results The apoptosis rate of CD4+CD25+Tregs in anti-CD3/CD28 group was 33.70± 3.06%,which was significantly higher than that in control group(12.84±0.84%).Also,apoptosis rate of CD4 CD25+Tregs in anti-CD3/CD28+LPS+Xuebijing injection group(45.13±2.70%)was much higher than that in anti-CD3/CD28+IPS group(29.41 ± 1.63%,P＜0.01).The expression of Foxp3 as well as CTLA-4,and the secretion of IL-10 were markedly decreased along with increases in the apoptosis rates.Compared with control group(54.48%),the mean inhibitory rate of Teff proliferative activity in response to Con A was significantly decreased in anti-CD3/CD28+Xuebijing injection group(31.26%,P＜0.05),and it was markedly decreased in anti-CD3/CD28+LPS+Xuebijing injection group comaped to anti-CD3/CD28+LPS group(P＜0.01).In addition,IL-2 levels in the supernatant of anti-CD3/CD28+Xuebijing injection group and anti-CD3/CD28+LPS+Xuebijing injection group were significantly higher than those of anti-CD3/CD28+IPS group(P＜0.01).Conclusions The inhibitory activity of CD4+CD25+Tregs on Teff appears to be upregulated by IPS stimulation in vitro,and Xuebijing injection could markedly enhance apoptosis of CD4+CD25+Tregs,thereby improving suppressive immune function of Teff.
