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呼吸性酸中毒酸化预处理对缺血再灌注肺损伤中性粒细胞炎性浸润的影响
作者:曲良超 赵娜 李倩倩 武勇强 莫非 
单位:330006 南昌 南昌大学第一附属医院麻醉科(曲良超、赵娜、李倩倩、武勇强、莫非) 
关键词:呼吸性酸中毒 预处理 缺血再灌注肺损伤 中性粒细胞 
分类号:R655.3
出版年,卷(期):页码:2019,44(10):843-850
摘要:

 [摘要]  目的  探讨呼吸性酸中毒酸化预处理对大鼠缺血再灌注(IR)肺损伤的作用及机制。方法  雄性SD大鼠48 只,体重250~300 g,随机分为6组,每组8只。对照组:开胸游离左肺门,未行阻断;假手术组:呼吸性酸中毒酸化预处理,未行阻断;IR组:阻断左肺门45 min后再灌注180 minAPC组:预处理+IRSB+IR组:腹腔单次注射MMP-9抑制剂SB-3CT 5 mg/kg30 min后行IRSB+APC组:MMP-9抑制剂+预处理+IR。实验结束后动脉放血处死动物,留取肺组织标本。采用HE染色观察大鼠肺组织病理学变化,免疫组化法检测肺组织中性粒细胞(PMN)的浸润情况,Western blotting检测NF-κBMMP-9蛋白的表达,免疫荧光染色检测NF-κBMMP-9蛋白的共表达情况。结果  各组大鼠肺组织病理学评分(对照组3.18±1.21,假手术组3.18±1.30IR13.85±5.62APC9.77±3.96SB+IR5.11±2.53SB+APC5.01±2.17)PMN浸润情况(各组IOD值分别为:对照组0.0015±0.0005,假手术组0.0014±0.0006IR0.0076±0.0012APC0.0057±0.0017SB+IR0.0035±0.001SB+APC0.0031±0.0011)NF-κBMMP-9蛋白的表达量(各组相对表达量分别为:对照组0.14±0.040.24±0.07,假手术组015±0.030.25±0.06IR0.56±0.091.09±0.17APC0.41±0.110.81±0.11SB+IR0.36±0.080.56±0.11SB+APC0.35±0.090.55±0.10)及二者共表达量的差异均有统计学意义(P<0.05)。各组以上指标间比较,假手术组与对照组差异无统计学意义(P>0.05)IR组、APC组、SB+IR组和SB+APC组均明显高于对照组和假手术组(P<0.05)APC组、SB+IR组和SB+APC组均明显低于IR(P<0.05)SB+IR组和SB+APC组均明显低于APC(P<0.05)SB+APC组与SB+IR组差异无统计学意义(P>0.05)结论  在体大鼠IR模型中,呼吸性酸中毒酸化预处理可减轻大鼠IR肺损伤及PMN浸润,其机制可能与抑制NF-κB/ MMP-9表达有关。

 [Abstracts]  Objective  To investigate the effect and mechanism of acidizing pretreatment in respiratory acidosis on ischemia-reperfusion (IR) lung injury in rats. Methods  Forty-eight male Sprague-Dawley rats, weighted 250-300 g, were randomly divided into 6 groups (8 each). Rats in control group: the left pulmonary hilum was dissociated without occlusion; in sham group: the respiratory rate was adjusted to maintain the partial pressure of end-tidal carbon dioxide reached 56-65 mmHg for 5 min, the left hilar not blocked; in IR group: the left pulmonary hilum was occluded for 45 min followed by reperfusion for 180 min; in group APC: done as in sham group and blocked the left hilar as in group IR; in group SB+IR: MMP-9 inhibitor SB-3CT 5 mg/kg was intraperitoneally injected, and then treated as in group IR 30 min after; in group SB+APC: done as in group APC, and then SB-3CT 5 mg/kg was injected 30 min after. Rats in all the 6 groups were sacrificed by arterial bleeding when the experiments ended, and lung tissues were taken. HE staining was performed to observe the pathological changes of lung tissues, and immunohistochemistry was used to detect the infiltration of neutrophils (PMN) in lung tissue. Western blotting was used to detect the expression of NF- κB and MMP-9 proteins. Immunofluorescence staining was used to detect the co-expression of NF-κB and MMP-9 proteins. Results  Statistically significant differences existed in each group (P<0.05) in the pathological scores of lung tissue (group control 3.18±1.21; sham group 3.18±1.30; group IR 13.85±5.62; group APC 9.77±3.96; group SB+IR 5.11±2.53; group SB+APC 5.01±2.17), PMN infiltration (IOD values of each group were: group control 0.0015±0.0005; group sham 0.0014±0.0006; group IR 0.0076±0.0012; group APC 0.0057±0.0017; group SB+IR 0.0035±0.001; group SB+APC 0.0031±0.0011), NF-κB and MMP-9 protein expression levels (relative expression values of each group were: control group 0.14±0.04, 0.24±0.07; group sham 015±0.03, 0.25±0.06; group IR 0.56±0.09, 1.09±0.17; group APC 0.41±0.11, 0.81±0.11; group SB+IR 0.36±0.08, 0.56±0.11; group SB+APC 0.35±0.09, 0.55±0.10), and the co-expression of NF-κB and MMP-9 proteins. Multiple comparison results of the above indicators between groups displayed that no significant difference existed between group sham andl group contro (P>0.05). The above indicators were significantly higher in group IR, group APC, group SB+IR and group SB+APC than those in control group and sham group (P<0.05); and were significantly lower in group APC, group SB+IR and group SB+APC than that in group IR (P<0.05); and were significantly lower in group SB+IR and group SB+APC than that in group APC (P<0.05); and no significant difference was found between group SB+APC and group SB+IR (P>0.05). Conclusions  In the IR rat model, acidizing pretreatment in respiratory acidosis can alleviate PMN infiltration and IR lung injury. The mechanism may be related to inhibition of NF-κB/MMP-9 protein expression.

基金项目:
江西省科技厅重点项目(20171BBG70054、20181BBG78022)
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