中华危重病急救医学
中華危重病急救醫學
중화위중병급구의학
Chinese Critical Care Medicine
2014年
5期
289-293
,共5页
戴惠军%潘灵辉%林飞%葛万运%李玮%贺盛
戴惠軍%潘靈輝%林飛%葛萬運%李瑋%賀盛
대혜군%반령휘%림비%갈만운%리위%하성
肺泡巨噬细胞%机械通气%呼吸机相关性肺损伤%Toll样受体9%髓样分化因子88
肺泡巨噬細胞%機械通氣%呼吸機相關性肺損傷%Toll樣受體9%髓樣分化因子88
폐포거서세포%궤계통기%호흡궤상관성폐손상%Toll양수체9%수양분화인자88
Alveolar macrophage%Mechanical ventilation%Ventilator-induced lung injury%Toll-like receptor 9%Myeloid differentiation factor 88
目的 探讨肺泡巨噬细胞Toll样受体9(TLR9)-髓样分化因子88(MyD88)信号通路在呼吸机相关性肺损伤(VILI)中的作用.方法 清洁级SD大鼠30只,按随机数字表法将大鼠分为3组,每组10只.A组保留自主呼吸作为对照;B组给予正常潮气量(VT,7 mL/kg)机械通气4h;C组给予大VT(40 mL/kg)机械通气4h.机械通气结束时,透射电镜下观察大鼠Ⅱ型肺泡上皮细胞(AECⅡ)超微结构改变;测定肺组织湿/干质量(W/D)比值以及支气管肺泡灌洗液(BALF)中总蛋白、白细胞介素(IL-6、IL-1β)的浓度;用蛋白质免疫印迹试验(Western Blot)和逆转录-聚合酶链反应(RT-PCR)分别检测肺泡巨噬细胞TLR9、MyD88、核转录因子-κB(NF-κB)的蛋白及其mRNA表达.结果 A、B组大鼠AECⅡ细胞超微结构基本正常;C组AECⅡ胞质内板层小体、细胞膜、细胞核中的染色质及微绒毛等均有不同程度的损伤性改变.C组较A组、B组肺组织W/D比值(5.54±0.17比4.58±0.17、4.69±0.16),BALF中总蛋白(g/L:6.33±0.61比0.45±0.05、0.47±0.04)、IL-6(μg/L:1.989±0.103比1.033±0.061、1.010±0.069)、IL-1β(ng/L:2.79±0.25比1.05±0.15、1.23±0.22),肺泡巨噬细胞TLR9、MyD88、NF-κB的蛋白表达[TLR9(A值):0.770±0.042比0.300±0.027、0.310±0.037,MyD88(A值):0.950±0.091比0.560±0.082、0.580±0.084,NF-κB(A值):1.020±0.076比0.740±0.052、0.700±0.076]均明显升高,差异有统计学意义(P＜0.05或P＜0.01).B组肺泡巨噬细胞TLR9、MyD88、NF-κB的mRNA表达分别是A组的(1.13±0.32)倍、(1.18±0.33)倍、(1.11±0.22)倍,差异均无统计学意义(均P＞0.05);C组肺泡巨噬细胞TLR9、MyD88、NF-κB的mRNA表达分别是A组的(8.66±0.69)倍、(6.41±0.53)倍、(5.29±0.71)倍,差异均有统计学意义(均P＜0.01).结论 肺泡巨噬细胞TLR9-MyD88信号通路参与并介导了机械通气所致的肺损伤.
目的 探討肺泡巨噬細胞Toll樣受體9(TLR9)-髓樣分化因子88(MyD88)信號通路在呼吸機相關性肺損傷(VILI)中的作用.方法 清潔級SD大鼠30隻,按隨機數字錶法將大鼠分為3組,每組10隻.A組保留自主呼吸作為對照;B組給予正常潮氣量(VT,7 mL/kg)機械通氣4h;C組給予大VT(40 mL/kg)機械通氣4h.機械通氣結束時,透射電鏡下觀察大鼠Ⅱ型肺泡上皮細胞(AECⅡ)超微結構改變;測定肺組織濕/榦質量(W/D)比值以及支氣管肺泡灌洗液(BALF)中總蛋白、白細胞介素(IL-6、IL-1β)的濃度;用蛋白質免疫印跡試驗(Western Blot)和逆轉錄-聚閤酶鏈反應(RT-PCR)分彆檢測肺泡巨噬細胞TLR9、MyD88、覈轉錄因子-κB(NF-κB)的蛋白及其mRNA錶達.結果 A、B組大鼠AECⅡ細胞超微結構基本正常;C組AECⅡ胞質內闆層小體、細胞膜、細胞覈中的染色質及微絨毛等均有不同程度的損傷性改變.C組較A組、B組肺組織W/D比值(5.54±0.17比4.58±0.17、4.69±0.16),BALF中總蛋白(g/L:6.33±0.61比0.45±0.05、0.47±0.04)、IL-6(μg/L:1.989±0.103比1.033±0.061、1.010±0.069)、IL-1β(ng/L:2.79±0.25比1.05±0.15、1.23±0.22),肺泡巨噬細胞TLR9、MyD88、NF-κB的蛋白錶達[TLR9(A值):0.770±0.042比0.300±0.027、0.310±0.037,MyD88(A值):0.950±0.091比0.560±0.082、0.580±0.084,NF-κB(A值):1.020±0.076比0.740±0.052、0.700±0.076]均明顯升高,差異有統計學意義(P＜0.05或P＜0.01).B組肺泡巨噬細胞TLR9、MyD88、NF-κB的mRNA錶達分彆是A組的(1.13±0.32)倍、(1.18±0.33)倍、(1.11±0.22)倍,差異均無統計學意義(均P＞0.05);C組肺泡巨噬細胞TLR9、MyD88、NF-κB的mRNA錶達分彆是A組的(8.66±0.69)倍、(6.41±0.53)倍、(5.29±0.71)倍,差異均有統計學意義(均P＜0.01).結論 肺泡巨噬細胞TLR9-MyD88信號通路參與併介導瞭機械通氣所緻的肺損傷.
목적 탐토폐포거서세포Toll양수체9(TLR9)-수양분화인자88(MyD88)신호통로재호흡궤상관성폐손상(VILI)중적작용.방법 청길급SD대서30지,안수궤수자표법장대서분위3조,매조10지.A조보류자주호흡작위대조;B조급여정상조기량(VT,7 mL/kg)궤계통기4h;C조급여대VT(40 mL/kg)궤계통기4h.궤계통기결속시,투사전경하관찰대서Ⅱ형폐포상피세포(AECⅡ)초미결구개변;측정폐조직습/간질량(W/D)비치이급지기관폐포관세액(BALF)중총단백、백세포개소(IL-6、IL-1β)적농도;용단백질면역인적시험(Western Blot)화역전록-취합매련반응(RT-PCR)분별검측폐포거서세포TLR9、MyD88、핵전록인자-κB(NF-κB)적단백급기mRNA표체.결과 A、B조대서AECⅡ세포초미결구기본정상;C조AECⅡ포질내판층소체、세포막、세포핵중적염색질급미융모등균유불동정도적손상성개변.C조교A조、B조폐조직W/D비치(5.54±0.17비4.58±0.17、4.69±0.16),BALF중총단백(g/L:6.33±0.61비0.45±0.05、0.47±0.04)、IL-6(μg/L:1.989±0.103비1.033±0.061、1.010±0.069)、IL-1β(ng/L:2.79±0.25비1.05±0.15、1.23±0.22),폐포거서세포TLR9、MyD88、NF-κB적단백표체[TLR9(A치):0.770±0.042비0.300±0.027、0.310±0.037,MyD88(A치):0.950±0.091비0.560±0.082、0.580±0.084,NF-κB(A치):1.020±0.076비0.740±0.052、0.700±0.076]균명현승고,차이유통계학의의(P＜0.05혹P＜0.01).B조폐포거서세포TLR9、MyD88、NF-κB적mRNA표체분별시A조적(1.13±0.32)배、(1.18±0.33)배、(1.11±0.22)배,차이균무통계학의의(균P＞0.05);C조폐포거서세포TLR9、MyD88、NF-κB적mRNA표체분별시A조적(8.66±0.69)배、(6.41±0.53)배、(5.29±0.71)배,차이균유통계학의의(균P＜0.01).결론 폐포거서세포TLR9-MyD88신호통로삼여병개도료궤계통기소치적폐손상.
Objective To investigate the role of Toll-like receptor9 (TLR9)-myeloid differentiation factor 88 (MyD88) signal pathway in alveolar macrophages in ventilator-induced lung injury (VILI).Methods 30 adult male Sprague-Dawley (SD) rats were randomly assigned to three groups (with 10 rats in each group).Group A was the control group,with spontaneous respiration after tracheostomy.Rats in group B received mechanical ventilation for 4 hours with normal tidal volume (VT) 7 ml/kg after tracheostomy,and group C rats received mechanical ventilation with VT 40 ml/kg for 4 hours.After termination of ventilation,examination with transmission electron microscopy was performed to observe the ultrastructure changes in alveolar epithelial cell type Ⅱ (AEC Ⅱ) of the lung.Lung wet/dry ratios (W/D) and total protein concentration,the concentration of interleukins (IL-6 and IL-1 β) in bronchoalveolar lavage fluid (BALF) were determined.The protein and mRNA expressions of TLR9,MyD88 and nuclear factor-κB (NF-κB) in alveolar macrophages were assayed by Western Blot and real-time reverse transcription-polymerase chain reaction (RT-PCR).Results The ultrastructure of AEC Ⅱ in the group A and group B was almost normal,whereas the chromatin of the nuclei,the lamellar corpuscles in the cytoplasm,the cell membrane and the microvilli of the AEC Ⅱ in the group C showed injurious changes in various degrees.When the group C was compared with the group A and the group B,it was shown that the W/D ratios (5.54 ± 0.17 vs.4.58 ± 0.17,4.69 ± 0.16) and total protein concentration (g/L:6.33 ± 0.61 vs.0.45 ± 0.05,0.47 ± 0.04),IL-6 (μg/L:1.989 ± 0.103 vs.1.033 ± 0.061,1.010 ± 0.069) and IL-lβ (ng/L:2.79 ±0.25 vs.1.05 ±0.15,1.23 ±0.22) in BALF,the protein expressions of TLR9,MyD88 and NF-κB [TLR9 (A value):0.770 ±0.042 vs.0.300 ±0.027,0.310 ±0.037; MyD88 (A value):0.950 ±0.091 vs.0.560 ±0.082,0.580±0.084; NF-κB(A value):1.020 ±0.076 vs.0.740 ±0.052,0.700 ±0.076] in alveolar macrophages were all increased significantly,and all of which showed significant difference (P＜0.05 or P＜0.01).The mRNA levels of TLR9,MyD88 and NF-κB in alveolar macrophages in the group B were (1.13 ± 0.32),(1.18 ± 0.33),and (1.11 ± 0.22) folds of those of the group A,respectively,but there were no significant differences (all P＞0.05).While the mRNA levels of TLR9,MyD88 and NF-κB of alveolar macrophages in the group C were (8.66 ± 0.69),(6.41 ± 0.53) and (5.29 ± 0.71) folds of those of the group A,respectively,and all of them showed significant difference (all P＜0.01).Conclusion TLR9-MyD88 signaling in alveolar macrophages plays a role in pathogenesis of VILI.
