中华烧伤杂志
中華燒傷雜誌
중화소상잡지
16
2013年
3期
261-266
,共6页
邵义如%申捷%李卫%袁震%何岱昆
邵義如%申捷%李衛%袁震%何岱昆
소의여%신첩%리위%원진%하대곤
烧伤,化学%烧伤,吸入性%光气%肺损伤%基质金属蛋白酶9%丝裂原活化蛋白激酶
燒傷,化學%燒傷,吸入性%光氣%肺損傷%基質金屬蛋白酶9%絲裂原活化蛋白激酶
소상,화학%소상,흡입성%광기%폐손상%기질금속단백매9%사렬원활화단백격매
Burns,chemical%Burns,inhalation%Phosgene%Lung injury%Matrix metalloproteinase 9%Mitogen-activated protein kinase
目的 探讨磷酸化MAPK[磷酸化细胞外信号调节蛋白激酶1/2(p-ERK1/2)、磷酸化蛋白激酶p38(p-p38)、磷酸化c-Jun氨基末端激酶(p-JNK)]表达在光气吸入所致肺损伤中的作用及其与基质金属蛋白酶9(MMP-9)的关系. 方法 选取30只雄性Wistar大鼠,按随机数字表法分为空气对照组(简称C组)、光气吸入组(简称P组)、PD98059(ERK1/2特异性阻断剂)组、SB203580(p38特异性阻断剂)组和SP600125(JNK特异性阻断剂)组,每组6只.计数支气管肺泡灌洗液(BALF)中性粒细胞,测定肺湿干比,ELISA法检测血清炎症因子TNF-α、IL-1β、IL-6、IL-8的表达水平,蛋白质印迹法检测各组肺组织中p-ERK1/2、p-p38、p-JNK、MMP-9的蛋白表达,实时荧光定量PCR法检测各组肺组织中MMP-9 mRNA表达.组间整体比较采用单因素方差分析,两两比较采用SNK法. 结果 与C组[中性粒细胞数为(2.0±0.7) ×104个/mL,肺湿干比为3.7±0.6]相比,P组BALF中性粒细胞数[(10.7±1.4) ×104个/mL]增多,肺湿干比(7.6±0.4)升高；而SB203580组、SP600125组BALF中中性粒细胞数分别为(8.3±1.1)×104、(7.9±1.3)×104个/mL,肺湿干比各为6.1±1.4、6.1±0.9,较P组均显著降低(P值均小于0.01).P组炎症因子TNF-α、IL-1β、IL-6、IL-8的表达较C组升高,而SP600125组和SB203580组的表达水平较P组有不同程度降低,但仍高于C组,差异均有统计学意义(P ＜0.05或P＜0.01).蛋白质印迹法结果显示,P组p-p38、p-JNK的表达量分别是1.19±0.22、1.43±0.14,较C组的0.76±0.06、0.74±0.05明显增强；PD98059组、SB203580组、SP600125组p-ERK1/2、p-p38、p-JNK蛋白表达量各为0.47±0.05、0.88±0.07、0.91±0.07,与P组比较均明显降低(P＜0.05或P＜0.01).P组MMP-9蛋白表达量(2.23±0.18)及mRNA表达量(4.93±0.12)分别较C组MMP-9蛋白表达量(1.26±0.14)和mRNA表达量(1.80±0.03)明显增强；SB203580组MMP-9蛋白表达量(1.58±0.14)和MMP-9 mRNA表达量(2.96±0.29)、SP600125组MMP-9蛋白表达量(1.55±0.30)、MMP-9 mRNA表达量(3.00±0.13)均低于P组,差异有统计学意义(P ＜0.05或P＜0.01). 结论 光气吸入可能激活MAPK信号蛋白通路,通过其活性形式p-p38、p-JNK蛋白表达增加,在光气吸入性肺损伤中发挥作用,其机制与上调MMP-9的表达相关.
目的 探討燐痠化MAPK[燐痠化細胞外信號調節蛋白激酶1/2(p-ERK1/2)、燐痠化蛋白激酶p38(p-p38)、燐痠化c-Jun氨基末耑激酶(p-JNK)]錶達在光氣吸入所緻肺損傷中的作用及其與基質金屬蛋白酶9(MMP-9)的關繫. 方法 選取30隻雄性Wistar大鼠,按隨機數字錶法分為空氣對照組(簡稱C組)、光氣吸入組(簡稱P組)、PD98059(ERK1/2特異性阻斷劑)組、SB203580(p38特異性阻斷劑)組和SP600125(JNK特異性阻斷劑)組,每組6隻.計數支氣管肺泡灌洗液(BALF)中性粒細胞,測定肺濕榦比,ELISA法檢測血清炎癥因子TNF-α、IL-1β、IL-6、IL-8的錶達水平,蛋白質印跡法檢測各組肺組織中p-ERK1/2、p-p38、p-JNK、MMP-9的蛋白錶達,實時熒光定量PCR法檢測各組肺組織中MMP-9 mRNA錶達.組間整體比較採用單因素方差分析,兩兩比較採用SNK法. 結果 與C組[中性粒細胞數為(2.0±0.7) ×104箇/mL,肺濕榦比為3.7±0.6]相比,P組BALF中性粒細胞數[(10.7±1.4) ×104箇/mL]增多,肺濕榦比(7.6±0.4)升高；而SB203580組、SP600125組BALF中中性粒細胞數分彆為(8.3±1.1)×104、(7.9±1.3)×104箇/mL,肺濕榦比各為6.1±1.4、6.1±0.9,較P組均顯著降低(P值均小于0.01).P組炎癥因子TNF-α、IL-1β、IL-6、IL-8的錶達較C組升高,而SP600125組和SB203580組的錶達水平較P組有不同程度降低,但仍高于C組,差異均有統計學意義(P ＜0.05或P＜0.01).蛋白質印跡法結果顯示,P組p-p38、p-JNK的錶達量分彆是1.19±0.22、1.43±0.14,較C組的0.76±0.06、0.74±0.05明顯增彊；PD98059組、SB203580組、SP600125組p-ERK1/2、p-p38、p-JNK蛋白錶達量各為0.47±0.05、0.88±0.07、0.91±0.07,與P組比較均明顯降低(P＜0.05或P＜0.01).P組MMP-9蛋白錶達量(2.23±0.18)及mRNA錶達量(4.93±0.12)分彆較C組MMP-9蛋白錶達量(1.26±0.14)和mRNA錶達量(1.80±0.03)明顯增彊；SB203580組MMP-9蛋白錶達量(1.58±0.14)和MMP-9 mRNA錶達量(2.96±0.29)、SP600125組MMP-9蛋白錶達量(1.55±0.30)、MMP-9 mRNA錶達量(3.00±0.13)均低于P組,差異有統計學意義(P ＜0.05或P＜0.01). 結論 光氣吸入可能激活MAPK信號蛋白通路,通過其活性形式p-p38、p-JNK蛋白錶達增加,在光氣吸入性肺損傷中髮揮作用,其機製與上調MMP-9的錶達相關.
목적 탐토린산화MAPK[린산화세포외신호조절단백격매1/2(p-ERK1/2)、린산화단백격매p38(p-p38)、린산화c-Jun안기말단격매(p-JNK)]표체재광기흡입소치폐손상중적작용급기여기질금속단백매9(MMP-9)적관계. 방법 선취30지웅성Wistar대서,안수궤수자표법분위공기대조조(간칭C조)、광기흡입조(간칭P조)、PD98059(ERK1/2특이성조단제)조、SB203580(p38특이성조단제)조화SP600125(JNK특이성조단제)조,매조6지.계수지기관폐포관세액(BALF)중성립세포,측정폐습간비,ELISA법검측혈청염증인자TNF-α、IL-1β、IL-6、IL-8적표체수평,단백질인적법검측각조폐조직중p-ERK1/2、p-p38、p-JNK、MMP-9적단백표체,실시형광정량PCR법검측각조폐조직중MMP-9 mRNA표체.조간정체비교채용단인소방차분석,량량비교채용SNK법. 결과 여C조[중성립세포수위(2.0±0.7) ×104개/mL,폐습간비위3.7±0.6]상비,P조BALF중성립세포수[(10.7±1.4) ×104개/mL]증다,폐습간비(7.6±0.4)승고；이SB203580조、SP600125조BALF중중성립세포수분별위(8.3±1.1)×104、(7.9±1.3)×104개/mL,폐습간비각위6.1±1.4、6.1±0.9,교P조균현저강저(P치균소우0.01).P조염증인자TNF-α、IL-1β、IL-6、IL-8적표체교C조승고,이SP600125조화SB203580조적표체수평교P조유불동정도강저,단잉고우C조,차이균유통계학의의(P ＜0.05혹P＜0.01).단백질인적법결과현시,P조p-p38、p-JNK적표체량분별시1.19±0.22、1.43±0.14,교C조적0.76±0.06、0.74±0.05명현증강；PD98059조、SB203580조、SP600125조p-ERK1/2、p-p38、p-JNK단백표체량각위0.47±0.05、0.88±0.07、0.91±0.07,여P조비교균명현강저(P＜0.05혹P＜0.01).P조MMP-9단백표체량(2.23±0.18)급mRNA표체량(4.93±0.12)분별교C조MMP-9단백표체량(1.26±0.14)화mRNA표체량(1.80±0.03)명현증강；SB203580조MMP-9단백표체량(1.58±0.14)화MMP-9 mRNA표체량(2.96±0.29)、SP600125조MMP-9단백표체량(1.55±0.30)、MMP-9 mRNA표체량(3.00±0.13)균저우P조,차이유통계학의의(P ＜0.05혹P＜0.01). 결론 광기흡입가능격활MAPK신호단백통로,통과기활성형식p-p38、p-JNK단백표체증가,재광기흡입성폐손상중발휘작용,기궤제여상조MMP-9적표체상관.
Objective To investigate the effects of phosphorylated mitogen-activated protein kinases (MAPK),including the phosphorylated extracellular signal-regulated protein kinase 1/2 (p-ERK1/2),the phosphorylated protein p38 (p-p38),the phosphorylated c-Jun N-terminal kinase (p-JNK),on phosgene inhalation-induced lung injury and its relationship with matrix metalloproteinase 9 (MMP-9).Methods According to the random number table,30 male Wistar rats were divided into air control group (C),phosgene inhalation group (P),PD98059 (specific inhibitor of ERK1/2) group,SB203580 (specific inhibitor of p38) group,and SP600125 (specific inhibitor of JNK) group,with 6 rats in each group.The number of neutrophils in the bronchoalveolar lavage fluid (BALF) was counted and the lung wet-dry ratio (W/D) was examined.The serum levels of inflammatory factors TNF-α,IL-1β,IL-6,and IL-8 were determined with ELISA.The protein expressions of p-ERK1/2,p-p38,p-JNK,and MMP-9 in lung tissue were detected with Western blotting.The mRNA level of MMP-9 in lung tissuc was detected with real-time fluorescence quantitative PCR.Data were processed with one-way analysis of variance (among groups) and SNK method (paired comparison).Results Compared with those of group C [respectively (2.0 ± 0.7) × 104/mL and 3.7 ± 0.6],the number of neutrophils and W/D of group P [respectively (10.7 ± 1.4) × 104/mL and 7.6 ± 0.4] were increased.The number of neutrophils in group SB203580 and group SP600125 was respectively (8.3 ± 1.1) × 104,(7.9 ± 1.3) × 104/mL,with W/D respectively 6.1 ± 1.4,6.1 ± 0.9,all of which decreased as compared with those of group P (with P values all below 0.01).Compared with those of group C,the levels of TNF-a,IL-1 β,IL-6,and IL-8 of group P were increased,but decreased in group SB203580 and group SP600125 compared with that of group P,though still higher than those of group C,and the differences were statistically significant (P ＜0.05 or P ＜ 0.01).Protein quantities of p-p38 and p-JNK were higher in group P (respectively 1.19±0.22 and 1.43 ±0.14) than in group C (respectively0.76 ±0.06 and 0.74 ± 0.05).Compared with those of group P,the protein lcvcls of p-ERK1/2 (0.47 ± 0.05) in group PD98059,p-p38 (0.88 ±0.07) in group SB203580,and p-JNK (0.91 ±0.07) in group SP600125 were significantly reduced (P ＜ 0.05 or P ＜0.01).The protein and mRNA levels of MMP-9 were higher in group P (respectively 2.23 ±0.18 and 4.93 ±0.12) than in group C (respectively 1.26 ±0.14 and 1.80 ±0.03).The protein and mRNA levels of MMP-9 in group SB203580 (respectively 1.58 ±0.14 and 2.96 ±0.09) and group SP600125 (respectively 1.55 ±0.30 and 3.00 ±0.13) were lower than those in group P (P ＜ 0.05 or P ＜ 0.01).Conclusions The phosgene inhalation can activate the MAPK signaling protein pathway by increasing expressions of p-p38 and p-JNK,which lead to an up-regulation of MMP-9,and this may contribute to the phosgene inhalation-induced lung injury.
