氯沙坦干预急性肺损伤小鼠肺Th1和Th17的作用
록사탄간예급성폐손상소서폐Th1화Th17적작용
Losartan modulates T helper type 1 cells and T helper type 17 cells-mediated responses in a mouse model of lipopolysaccharide-induced acute lung injury
  • 万方数据
    中华内科杂志 중화내과잡지
    CHINESE JOURNAL OF INTERNAL MEDICINE
    2014年 10期 804-808 ,共5页

    刘军%张朋书%袈宏丽%于涛%刘玲%郭凤梅%黄英姿%杨毅%邱海波

    류군%장붕서%가굉려%우도%류령%곽봉매%황영자%양의%구해파

    急性肺损伤%T淋巴细胞,辅助诱导%氯沙坦%炎症 急性肺損傷%T淋巴細胞,輔助誘導%氯沙坦%炎癥
    급성폐손상%T림파세포,보조유도%록사탄%염증
    Acute lung injury%T-Lymphocytes,helper-inducer%Losartan%Inflammation
    目的 探讨氯沙坦干预急性肺损伤(ALI)小鼠肺Th1和Th17的作用及机制.方法 小鼠随机分为:(1)对照组;(2) ALI组:气管内注射脂多糖2 mg/kg复制ALI模型;(3)氯沙坦干预组:气管内注射脂多糖2 mg/kg前30 min腹腔内注射氯沙坦15 mg/kg.分别于24 h、48 h处死小鼠,取肺组织.光镜观察肺组织病理改变,进行Smith肺损伤半定量评分,测肺湿重/体重(LW/BW).实时定量PCR检测肺组织转录因子T细胞上的T盒(T-bet)、鸟嘌呤腺嘌呤胸腺嘧啶腺嘌呤序列结合蛋白3(GATA-3)、维甲酸相关孤独受体γt(RORγt)mRNA表达,ELISA测肺组织匀浆中IL-6、IFNγ IL-4和IL-17水平.结果 (1)与对照组比,ALI组小鼠肺LW/BW、Smith肺损伤半定量评分和肺组织匀浆中IL-6水平显著升高(P＜0.05).与ALI组比,氯沙坦干预组肺LW/BW、Smith肺损伤半定量评分、肺组织匀浆中IL-6水平降低(P＜0.05).(2)与对照组比,ALI组小鼠肺组织T-betmRNA (24h:2.4±0.4,48 h:6.8±0.7)、RORγt mRNA(24 h:1.7±0.4;48 h:2.9±0.8)表达增高(P＜0.05),但各组GATA-3 mRNA表达差异无统计学意义(P＞0.05).ALI组肺组织匀浆中IFγ水平[24 h:(745±90) pg/mg比(182 ±35) pg/mg,P＜0.05;48 h:(752±122) pg/mg比(177 ±61)pg/mg,P＜0.05]、IL-4水平[24 h:(206±37) pg/mg比(107±71) pg/mg,P＜0.05;48 h:(267±94)pg/mg比(94±53) pg/mg,P＜0.05]、IL-17水平高于对照组[24 h:(486±94) pg/mg比(122±30)pg/mg,P＜0.05;48 h:(518 ±124) pg/mg比(120±40) pg/mg,P＜0.05].(3)与ALI组比,氯沙坦干预组肺组织T-bet mRNA(24 h:1.8±0.4比2.4±0.4,P＜0.05;48 h:4.2±1.8比6.8±0.7,P＜0.05)、RORγt mRNA表达下调(24 h:1.3±0.2比1.7±0.4,P＜0.05;48 h:1.5±0.5比2.9±0.8,P＜0.05);肺组织匀浆中IFNγ水平[24 h:(503±136) pg/mg比(745±90) pg/mg,P＜0.05;48 h:(553±134) pg/mg比(752±122) pg/mg,P＜0.05]、IL-17水平降低[24 h:(374±106) pg/mg比(486±94) pg/mg,P＜0.05;48 h:(260±98) pg/mg比(518±124) pg/mg,P＜0.05],但IL-4水平差异无统计学意义(P＞0.05).结论 ALI时,肺Th1及Th17型极化反应可能参与ALI病程中的免疫失衡.氯沙坦对ALI具有部分保护性效应,可能通过抑制Th1及Th17型极化反应而对ALI具有抗炎和免疫调节反应.
    목적 탐토록사탄간예급성폐손상(ALI)소서폐Th1화Th17적작용급궤제.방법 소서수궤분위:(1)대조조;(2) ALI조:기관내주사지다당2 mg/kg복제ALI모형;(3)록사탄간예조:기관내주사지다당2 mg/kg전30 min복강내주사록사탄15 mg/kg.분별우24 h、48 h처사소서,취폐조직.광경관찰폐조직병리개변,진행Smith폐손상반정량평분,측폐습중/체중(LW/BW).실시정량PCR검측폐조직전록인자T세포상적T합(T-bet)、조표령선표령흉선밀정선표령서렬결합단백3(GATA-3)、유갑산상관고독수체γt(RORγt)mRNA표체,ELISA측폐조직균장중IL-6、IFNγ IL-4화IL-17수평.결과 (1)여대조조비,ALI조소서폐LW/BW、Smith폐손상반정량평분화폐조직균장중IL-6수평현저승고(P＜0.05).여ALI조비,록사탄간예조폐LW/BW、Smith폐손상반정량평분、폐조직균장중IL-6수평강저(P＜0.05).(2)여대조조비,ALI조소서폐조직T-betmRNA (24h:2.4±0.4,48 h:6.8±0.7)、RORγt mRNA(24 h:1.7±0.4;48 h:2.9±0.8)표체증고(P＜0.05),단각조GATA-3 mRNA표체차이무통계학의의(P＞0.05).ALI조폐조직균장중IFγ수평[24 h:(745±90) pg/mg비(182 ±35) pg/mg,P＜0.05;48 h:(752±122) pg/mg비(177 ±61)pg/mg,P＜0.05]、IL-4수평[24 h:(206±37) pg/mg비(107±71) pg/mg,P＜0.05;48 h:(267±94)pg/mg비(94±53) pg/mg,P＜0.05]、IL-17수평고우대조조[24 h:(486±94) pg/mg비(122±30)pg/mg,P＜0.05;48 h:(518 ±124) pg/mg비(120±40) pg/mg,P＜0.05].(3)여ALI조비,록사탄간예조폐조직T-bet mRNA(24 h:1.8±0.4비2.4±0.4,P＜0.05;48 h:4.2±1.8비6.8±0.7,P＜0.05)、RORγt mRNA표체하조(24 h:1.3±0.2비1.7±0.4,P＜0.05;48 h:1.5±0.5비2.9±0.8,P＜0.05);폐조직균장중IFNγ수평[24 h:(503±136) pg/mg비(745±90) pg/mg,P＜0.05;48 h:(553±134) pg/mg비(752±122) pg/mg,P＜0.05]、IL-17수평강저[24 h:(374±106) pg/mg비(486±94) pg/mg,P＜0.05;48 h:(260±98) pg/mg비(518±124) pg/mg,P＜0.05],단IL-4수평차이무통계학의의(P＞0.05).결론 ALI시,폐Th1급Th17형겁화반응가능삼여ALI병정중적면역실형.록사탄대ALI구유부분보호성효응,가능통과억제Th1급Th17형겁화반응이대ALI구유항염화면역조절반응.
    Objective To assess the effect of losartan,angiotensin Ⅱ receptor type 1 (AT1) receptor antagonist,on the pulmonary T helper (Th) cell polarization response in acute lung injury (ALI) mice.Methods C57BL/6 mice were randomized into control group,ALI group and ALI + losartan group,which respectively were administrated with phosphate buffered saline (PBS),2 mg/kg lipopolvsaccharide (LPS) and 2 mg/kg LPS as well as 15 mg/kg losartan 30 minutes before intratracheal injection of LPS.Lung wet weight/body weight (LW/BW) was recorded to assess the severity of lung injury.The mRNA expression levels of T-box expressed in T cells (T-bet),GATA-binding protein-3 (GATA-3) and retinoidrelated orphan nuclear receptor γt (RORγt) were quantitatively measured by real-time polymerase chain reaction (RT-PCR).Meanwhile,interleukin 6 (IL-6),interferon γ (IFNγ),IL-4 and IL-17 in lung homogenates were assessed by enzyme-linked immunosorbent assay (ELISA).Results (1) LW/BW was significantly increased in ALI group compared with ALI+ losartan group.(2) Histologically,widespread interstitial thickening with edema,severe alveolar hemorrhage,and diffuse interstitial infiltration of inflammatory cells were observed in the ALI group.Whereas,losartan effectively attenuated the LPS-induced alveolar hemorrhage and leukocyte infiltration.(3) The levels of IL-6 in lung tissue were significantly enhanced in the LPS-induced ALI mice,while it markedly decreased in ALI+ losartan group.(4) The mRNA expression of T-bet and RORγt was up-regulated in ALI mice at 24 h and 48 h compared to control group (P ＜ 0.05).There was no significant difference in the expression of GATA-3.In addition,compared with ALI group,ALI mice with pretreatment of losartan resulted in significantly reduced mRNA expression of T-bet at 24 h and 48 h and RORγt mRNA expression at 48 h (all P ＜ 0.05).(5) Meanwhile,the levels of IFNγ,IL-4,IL-17 and IL-6 in lung tissue were significantly enhanced at 24 h and 48 h in the LPS-induced ALI mice.In addition,both IFNγ and IL-17 in lung tissue at 24 h and 48 h decreased significantly in losartan-pretreated mice compared with the ALI mice.However,the level of IL-4 in lungs was similar in ALI group and ALI + losartan group.Conclusions Losartan has a protective effect on LPS-induced ALI,which may be partly dependent on suppressions of Th1 and Th17 polarization response.
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