中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2014年
1期
58-62
,共5页
赵娜%倪媛媛%赵立青%吴震州%尹芝南
趙娜%倪媛媛%趙立青%吳震州%尹芝南
조나%예원원%조립청%오진주%윤지남
T淋巴细胞亚群%伴刀豆球蛋白A%肝损伤,急性%模型,动物
T淋巴細胞亞群%伴刀豆毬蛋白A%肝損傷,急性%模型,動物
T림파세포아군%반도두구단백A%간손상,급성%모형,동물
T-lymphocyte subsets%Coneanavalin A%Liver injury,acute%Models,animal
目的 探讨γδ T淋巴细胞对刀豆蛋白A (Con A)诱导的肝损伤的保护作用.方法 采用静脉注射Con A的方法建立小鼠急性肝损伤模型,通过监测血清ALT及细胞因子水平,评估小鼠肝脏损伤的程度.在不同的时间点处死小鼠后分离肝脏淋巴细胞,流式细胞术分析肝脏γδ T淋巴细胞的比例及其亚群比例变化情况.组间比较用t检验或单因素方差分析,P＜0.05为差异有统计学意义.结果 流式细胞分析结果表明,随着肝损伤程度的加重,小鼠肝脏γδ T淋巴细胞比例呈增高趋势.T淋巴细胞抗原受体δ-/-小鼠注射Con A后6、12、24、48、72 h的血清ALT水平分别为(6302.61±592.06) U/L、(6569.44±1060.98)U/L、(6514.29±757.26) U/L、(1262.61±558.07) U/L、(226.54±98.20) U/L,与野生型对照小鼠的(1319.26±355.48)U/L、(3415.53±343.90) U/L、(2062.73±365.67) U/L、(113.66±113.26) U/L、(42.35±21.51)U/L比较,t值分别为14.430、5.656、10.590、4.035和3.664,P值均＜0.05.同时,血清炎症因子干扰素γ、肿瘤坏死因子α及白细胞介素4在注射Con A后3h内的表达水平也迅速升高.对肝脏内γ δ T淋巴细胞两个主要亚群含量的分析结果显示,注射Con A后,Vγ4 γ δ T淋巴细胞占总γδ T淋巴细胞和肝脏总淋巴细胞的比例分别为25.26％±2.43％和0.66％±0.05％,与对照组的17.785％±2.95％和0.47％±0.07％相比,t值分别为-4.18和-5.11,P值均＜0.01；Vγl γδ T淋巴细胞占总γδ T淋巴细胞的比例为50.19％±5.52％,明显高于对照组的38.37％±6.10％(t=-3.213,P＜ 0.05),但Vγl γ δ T淋巴细胞占肝脏总淋巴细胞的比例没有明显变化(0.78％±0.25％对比0.76％±0.18％,t=-0.146,P＞0.05).回输Vγ4 γ δ T淋巴细胞组小鼠ALT水平为(5054.10±1748.51) U/L,与回输Vγ1 γδ T淋巴细胞组小鼠的(12333.56±663.54)U/L和只预先注射磷酸盐缓冲液对照组小鼠的(11904.18±1 363.46) U/L相比,差异有统计学意义(F=28.03,P＜0.01). 结论 γδ T淋巴细胞能对Con A诱导的肝损伤发挥保护作用,并且这种保护作用可能是通过Vγ4 γ δ T淋巴细胞亚群发挥作用.
目的 探討γδ T淋巴細胞對刀豆蛋白A (Con A)誘導的肝損傷的保護作用.方法 採用靜脈註射Con A的方法建立小鼠急性肝損傷模型,通過鑑測血清ALT及細胞因子水平,評估小鼠肝髒損傷的程度.在不同的時間點處死小鼠後分離肝髒淋巴細胞,流式細胞術分析肝髒γδ T淋巴細胞的比例及其亞群比例變化情況.組間比較用t檢驗或單因素方差分析,P＜0.05為差異有統計學意義.結果 流式細胞分析結果錶明,隨著肝損傷程度的加重,小鼠肝髒γδ T淋巴細胞比例呈增高趨勢.T淋巴細胞抗原受體δ-/-小鼠註射Con A後6、12、24、48、72 h的血清ALT水平分彆為(6302.61±592.06) U/L、(6569.44±1060.98)U/L、(6514.29±757.26) U/L、(1262.61±558.07) U/L、(226.54±98.20) U/L,與野生型對照小鼠的(1319.26±355.48)U/L、(3415.53±343.90) U/L、(2062.73±365.67) U/L、(113.66±113.26) U/L、(42.35±21.51)U/L比較,t值分彆為14.430、5.656、10.590、4.035和3.664,P值均＜0.05.同時,血清炎癥因子榦擾素γ、腫瘤壞死因子α及白細胞介素4在註射Con A後3h內的錶達水平也迅速升高.對肝髒內γ δ T淋巴細胞兩箇主要亞群含量的分析結果顯示,註射Con A後,Vγ4 γ δ T淋巴細胞佔總γδ T淋巴細胞和肝髒總淋巴細胞的比例分彆為25.26％±2.43％和0.66％±0.05％,與對照組的17.785％±2.95％和0.47％±0.07％相比,t值分彆為-4.18和-5.11,P值均＜0.01；Vγl γδ T淋巴細胞佔總γδ T淋巴細胞的比例為50.19％±5.52％,明顯高于對照組的38.37％±6.10％(t=-3.213,P＜ 0.05),但Vγl γ δ T淋巴細胞佔肝髒總淋巴細胞的比例沒有明顯變化(0.78％±0.25％對比0.76％±0.18％,t=-0.146,P＞0.05).迴輸Vγ4 γ δ T淋巴細胞組小鼠ALT水平為(5054.10±1748.51) U/L,與迴輸Vγ1 γδ T淋巴細胞組小鼠的(12333.56±663.54)U/L和隻預先註射燐痠鹽緩遲液對照組小鼠的(11904.18±1 363.46) U/L相比,差異有統計學意義(F=28.03,P＜0.01). 結論 γδ T淋巴細胞能對Con A誘導的肝損傷髮揮保護作用,併且這種保護作用可能是通過Vγ4 γ δ T淋巴細胞亞群髮揮作用.
목적 탐토γδ T림파세포대도두단백A (Con A)유도적간손상적보호작용.방법 채용정맥주사Con A적방법건립소서급성간손상모형,통과감측혈청ALT급세포인자수평,평고소서간장손상적정도.재불동적시간점처사소서후분리간장림파세포,류식세포술분석간장γδ T림파세포적비례급기아군비례변화정황.조간비교용t검험혹단인소방차분석,P＜0.05위차이유통계학의의.결과 류식세포분석결과표명,수착간손상정도적가중,소서간장γδ T림파세포비례정증고추세.T림파세포항원수체δ-/-소서주사Con A후6、12、24、48、72 h적혈청ALT수평분별위(6302.61±592.06) U/L、(6569.44±1060.98)U/L、(6514.29±757.26) U/L、(1262.61±558.07) U/L、(226.54±98.20) U/L,여야생형대조소서적(1319.26±355.48)U/L、(3415.53±343.90) U/L、(2062.73±365.67) U/L、(113.66±113.26) U/L、(42.35±21.51)U/L비교,t치분별위14.430、5.656、10.590、4.035화3.664,P치균＜0.05.동시,혈청염증인자간우소γ、종류배사인자α급백세포개소4재주사Con A후3h내적표체수평야신속승고.대간장내γ δ T림파세포량개주요아군함량적분석결과현시,주사Con A후,Vγ4 γ δ T림파세포점총γδ T림파세포화간장총림파세포적비례분별위25.26％±2.43％화0.66％±0.05％,여대조조적17.785％±2.95％화0.47％±0.07％상비,t치분별위-4.18화-5.11,P치균＜0.01；Vγl γδ T림파세포점총γδ T림파세포적비례위50.19％±5.52％,명현고우대조조적38.37％±6.10％(t=-3.213,P＜ 0.05),단Vγl γ δ T림파세포점간장총림파세포적비례몰유명현변화(0.78％±0.25％대비0.76％±0.18％,t=-0.146,P＞0.05).회수Vγ4 γ δ T림파세포조소서ALT수평위(5054.10±1748.51) U/L,여회수Vγ1 γδ T림파세포조소서적(12333.56±663.54)U/L화지예선주사린산염완충액대조조소서적(11904.18±1 363.46) U/L상비,차이유통계학의의(F=28.03,P＜0.01). 결론 γδ T림파세포능대Con A유도적간손상발휘보호작용,병차저충보호작용가능시통과Vγ4 γ δ T림파세포아군발휘작용.
Objective To investigate the role played by γδ T cells in acute liver injury using the concanavalin A (ConA)-induced liver injury mouse model.Methods Acute liver injury was induced by intravenous injection of 10 μg/g of ConA into male C57BL/6J mice with wild-type or T cell receptorgamma knockout (TCR δ-/-) genetic backgrounds.Mice injected with PBS alone served as negative controls.The degree of liver damage was assessed by measuring serum levels oftransaminase and cytokines at postinjection hours 3,6,12,24,48,and 72.The percentage ofγδ T cells and proportions of different subsets in liver lymphocytes were measured by flow cytometry.Results The TCR δ-/-mice showed significantly higher levels of the inflammatory cytokines IFN-γ,TNFct and IL-4 than the wild-type mice at post-injection hour 3.The percentage of liver γδ T cells increased with increased injury degree,and the extent of increase was significantly higher in the TCR δ-/-mice than the wild-type mice (post-injection hour 6:6302.61 ± 592.06 vs.1319.26 ± 355.48,12:6569.44 ± 1060.98 vs.3415.53 ± 343.90,24:6514.29 ± 757.26 vs.2062.73 ± 365.67,48:1262.61 ± 558.07 vs.113.66 ± 113.26,and 72:226.54 ± 98.20 vs.42.35 ± 21.51 U/L; allP ＜ 0.05).In addition,compared to the negative control mice,the ConA-induced mice showed a higher proportions ofVγ4 γδ T cells to total γδ T cells (17.78 ± 2.95 vs.25.26 ± 2.43) and to total liver lymphocytes (0.47 ± 0.07 vs.0.66 ± 0.05).Similarly,compared to the negative control mice,the ConA-induced mice showed a higher proportion of Vγl γδ T cells to total γδ T cells (38.37 ± 6.10 vs.50.19 ± 5.52) but the proportion to total liver lymphocytes was not significantly different among the groups (0.76 ± 0.18 vs.0.78 ± 0.25).Reinfusion of Vγ4 γδ T lymphocytes into TCR-/-mice led to lower serum ALT levels than reinfusion ofVγl γδ T lymphocytes (5054.10 ± 1748.51 vs.12333.56 ± 663.535 U/L).Conclusion γδ T cells play a protective role in ConA-induced liver injury and this effect maybe mediated by the Vγ4 γδ T cell subset.
