目的:探讨严重脓毒症幼猪生化指标、肺部病理损伤与免疫机制间的相互关系，以及参附注射液的干预作用。方法:将2～3月龄巴拿马香猪按随机数字表法分为假手术组（Sham组；静脉注射生理盐水）、脂多糖（LPS）致严重脓毒症模型组（LPS组；静脉注射LPS 1 mg/kg，0.5 mg·kgn -1·hn -1维持12 h）、参附注射液干预组（SF组；制模同时静脉注射参附注射液10 mL/kg，每日2次），每组5只。制模后48 h取血检测C-反应蛋白（CRP）、降钙素原（PCT）、氧合指数（PaOn 2/FiOn 2）、剩余碱（BE）、血乳酸（Lac）等生化指标；取外周血行流式细胞检测，分析中性粒细胞数目〔髓过氧化物酶阳性（MPOn +）〕及其激活亚群CD11bn + CD64n +、M1型巨噬细胞（CD80n + CD64n +）、CD4n +和CD8n + T细胞的变化；采用酶联免疫吸附试验（ELISA）检测血浆细胞因子水平；苏木素-伊红（HE）染色观察肺组织病理损伤；采用反转录-聚合酶链反应（RT-PCR）检测肺组织损伤相关分子及其受体、趋化因子、炎性因子、血管内皮相关分子、紧密连接蛋白的mRNA表达。n 结果:与Sham组比较，LPS组CRP、PCT、Lac水平明显升高，PaOn 2/FiOn 2、BE明显下降；外周血CD80n + CD64n +、CD11bn + CD64n +、MPOn +、CD4n +和CD8n + T细胞数、肿瘤坏死因子-α（TNF-α）水平明显降低，白细胞介素-10（IL-10）、内皮细胞生长因子（VEGF）水平明显升高；肺组织高迁移率族蛋白B1（HMGB1）、晚期糖基化终末产物受体（RAGE）、促血管生成素2/1（Ang2/Ang1）的mRNA表达均明显升高，Toll样受体9（TLR9）、趋化因子（CXCL9、CXCL10）、TNF-α、IL-27、内皮细胞TEK酪氨酸激酶2（TIE2）、紧密连接蛋白血管内皮-钙黏蛋白（VE-CAD）和封闭蛋白（Occludin）的mRNA表达均明显下降；HE染色显示，肺泡腔炎性细胞浸润渗出、肺泡实变及肺泡间质层明显增厚、肺气肿。与LPS组比较，SF组Lac明显下降（mmol/L：4.2±1.0比6.3±1.1，n P<0.05），BE、PaOn 2/FiOn 2明显升高〔BE（mmol/L）：-6.4±2.6比-11.6±2.5，PaOn 2/FiOn 2（mmHg，1 mmHg＝0.133 kPa）：180±36比105±35，均n P<0.05〕，CD80n + CD64n +、CD11bn + CD64n +、MPOn +细胞比例均明显上升〔CD80n + CD64n +：（7.13±2.01）%比（3.80±0.46）%，CD11bn + CD64n +：（8.33±2.55）%比（2.15±0.47）%，MPOn +：（21.22±2.33）%比（8.31±0.46）%，均n P<0.05〕；肺组织HMGB1、RAGE的mRNA表达有一定程度下降〔HMGB1 mRNA（2n -ΔΔCT）：1.81±0.45比2.23±0.85，RAGE mRNA（2n -ΔΔCT）：6.69±3.48比11.60±6.91，均n P<0.05〕，CXCL9和TIE2的mRNA表达有一定程度上升〔CXCL9 mRNA（2n -ΔΔCT）：1.06±0.63比0.50±0.12，TIE2 mRNA（2n -ΔΔCT）：1.42±0.68比0.27±0.16，均n P<0.05〕；肺组织病理改变明显减轻。n 结论:BE、Lac、PaOn 2/FiOn 2的加重，免疫耗竭、麻痹和无能，可能是导致严重脓毒症幼猪肺脏致死性损伤的重要因素，该损伤可能与HMGB1及其受体RAGE过度活化，TLR9和相应趋化因子及炎性因子受到抑制，导致血管内皮细胞损伤加重和紧密连接蛋白表达减少的毛细血管渗漏机制相关。参附注射液改善严重肺炎并脓毒症的免疫紊乱可能与上调外周血M1型巨噬细胞和激活中性粒细胞、抑制HMGB1及其受体RAGE表达、上调CXCL9和TIE2表达有关。n “,”Objective:To investigate the relationship between the biochemical parameters, the pulmonary pathologic injury and the immune mechanism of severe sepsis in infant porcine, and the intervention effect of Shenfu injection.Methods:Panamanian infant porcine (2-3 months old) were divided into sham operation group (Sham group; intravenous injection of normal saline), lipopolysaccharide (LPS) induced severe sepsis model group (LPS group; intravenous injection of LPS 1 mg/kg, and continuing at 0.5 mg·kgn -1·hn -1 for 12 hours), and Shenfu injection intervention group (SF group; intravenous injection of Shenfu injection 10 mL/kg at the same time of modeling, twice a day) according to the random number table method, with 5 in each group. Forty-eight hours after the challenge, the changes of C-reactive protein (CRP), procalcitonin (PCT), oxygenation index (PaOn 2/FiOn 2), base excess (BE), blood lactate (Lac) and other biochemical indexes were detected with blood sampling; the number of neutrophils [myeloperoxidase positive (MPOn +)] and their activated subsets CD11bn + CD64n +, M1 macrophages (CD80n + CD64n +), CD4n + and CD8n + T cells were analyzed in peripheral blood by flow cytometry. The levels of plasma cytokines were detected by enzyme linked immunosorbent assay (ELISA). The pathological damage of lung tissue was observed by hematoxylin-eosin (HE) staining. The mRNA expression of lung injury related molecules and their receptors, chemokines, cytokines, vascular endothelial related molecules and tight junction protein were detected by reverse transcription-polymerase chain reaction (RT-PCR).n Results:Compared with Sham group, the levels of CRP, PCT and Lac in LPS group significantly increased, and PaOn 2/FiOn 2 and BE significantly decreased. In the peripheral blood, CD80n + CD64n + macrophages, CD11bn + CD64n + and MPOn + neutrophils, CD4n +, CD8n + T cells and tumor necrosis factor-α (TNF-α) significantly decreased, but interleukin-10 (IL-10) and vascular endothelial cell growth factor (VEGF) significantly increased; the mRNA expressions of high mobility group protein B1 (HMGB1), receptor for advanced glycation end products (RAGE), angiopoietin 2/angiopoietin 1 (Ang2/Ang1) significantly increased, Toll like receptor 9 (TLR9), chemokines (CXCL9 and CXCL10), TNF-α, IL-27, Tek tyrosine kinase 2 (TIE2), vascular endothelial cadherin (VE-CAD) and Occludin significantly decreased in lung tissue. HE staining showed inflammatory cell infiltration and exudation in the alveolar cavity, alveoli consolidation, thickening of the alveolar interstitial layer and emphysema. Compared with LPS group, Lac in SF group significantly decreased (mmol/L: 4.2±1.0 vs. 6.3±1.1,n P < 0.05), while BE and PaO n 2/FiOn 2 significantly increased [BE (mmol/L): -6.4±2.6 vs. -11.6±2.5, PaOn 2/FiOn 2 (mmHg, 1 mmHg = 0.133 kPa): 180±36 vs. 105±35, both n P < 0.05]; the percentage of CD80 n + CD64n + macrophages, CD11bn + CD64n + and MPOn + neutrophils significantly increased [CD80n + CD64n +: (7.13±2.01)% vs. (3.80±0.46)%, CD11bn + CD64n +: (8.33±2.55)% vs. (2.15±0.47)%, MPOn +: (21.22±2.33)% vs. (8.31±0.46)%, all n P < 0.05]; the mRNA expressions of HMGB1 and RAGE decreased to some extent [HMGB1 mRNA (2 n -ΔΔCT): 1.81±0.45 vs. 2.23±0.85, RAGE mRNA (2n -ΔΔCT): 6.69±3.48 vs. 11.60±6.91, both n P < 0.05], the mRNA expressions of CXCL9 and TIE2 increased to a certain extent [CXCL9 mRNA (2 n -ΔΔCT): 1.06±0.63 vs. 0.50±0.12, TIE2 mRNA (2n -ΔΔCT): 1.42±0.68 vs. 0.27±0.16, both n P < 0.05]; the pathological damage of lung tissue were significantly alleviated.n Conclusions:The increased abnormality of BE, Lac, and PaOn 2/FiOn 2, the immune exhausting and paralysis may be important factors leading to the fatal lung injury in infant porcine with severe sepsis. The possible mechanism is that the excessive activation of HMGB1 and its receptor RAGE, the suppression of TLR9 and corresponding chemokine and inflammatory factors lead to increased endothelial damage and decreased tight connection, which in turn induces capillary leakage. The intervention of immune disorders by Shenfu injection in the severe pneumonia accompanied by sepsis may be related to elevated M1 macrophages and activated neutrophils in the peripheral blood, inhibition of HMGB1 and its receptor RAGE mRNA expression, and elevated CXCL9 and TIE2 expression.n