摘要
目的建立一种经肝门静脉注射制备肠源性脓毒症(GOS)大鼠模型。方法清洁级SD大鼠84只(250~300 g,12周龄),12只大鼠用于盲肠结扎并穿孔法(CLP)制备肠源性腹腔渗出液。72只大鼠采用随机数字法分为肝门静脉对照组(PV-S组)、肝门静脉渗液组(PV-E组)和下腔静脉渗液组(VC-E组),每组24只。PV-S组经肝门静脉注射0.9%氯化钠注射液(1 m L/只),PV-E组和VC-E组分别经肝门静脉和下腔静脉注射等量腹腔渗出液。各组大鼠再按腹腔渗液注射后1,2,4,6 h分为4个亚组。各亚组大鼠在相应时相点经腹主动脉采血,处死后取肺组织并收集肺泡灌洗液(BALF),测定血清和BALF中内毒素、肿瘤坏死因子-α(TNF-α)及白细胞介素-6(IL-6)含量,计算肺组织湿/干重(W/D)比值,观察肺形态学和病理形态学变化。结果 PV-E组在腹腔渗液注射液后1,2,4,6 h血清和BALF中内毒素、TNF-α及IL-6含量以及肺组织病理学评分、肺湿/干重比均显著高于同时相点PV-S组大鼠(P均<0.05)。与PV-E组比较,VC-E组在腹腔渗液注射液后1,2,4,6 h血清和BALF中内毒素水平均升高(P均<0.05),而血清和BALF中TNF-α、IL-6水平降低(P均<0.05)。PV-E组大鼠经肝门静脉注射腹腔渗出液后1,2,4,6 h肺组织W/D值(1 h:8.53±2.05;2 h:9.41±1.24;4 h:10.35±0.69;6 h:11.05±1.95)较同时相点的PV-S组(1 h:5.16±1.13;2 h:5.17±1.34;4 h:5.14±2.04;6 h:5.13±1.95)和VC-E组(1 h:6.71±0.73;2 h:7.65±1.32;4 h:8.40±0.43;6 h:9.41±1.87)均明显增大(P均<0.05)。与PV-S组比较,PV-E组肺组织明显肿胀,有广泛点、片状出血点及淤血;VC-E组肺组织病变程度较PV-E组轻。与PV-S组和VC-E组比较,PV-E组肺病理组织学结果显示肺泡腔中见大量中性粒细胞、单核细胞和红细胞浸润,肺泡壁破坏严重、肺间质明显水肿,各时相点病理学组织学评分高于PV-S组(1 h:5.32±1.35 vs 0.25±0.02;2 h:6.44±0.72 vs 0.23±0.04;4 h:7.52±1.45 vs 0.27±0.02;6 h:8.69±0.35 vs 0.22±0.05,P均<0.05)和VC-E组
Objective To establish a rat model of gut origin sepsis induced by portal vein injection.Methods Twelve rats of 84 SD rats( 250 ~ 300 g,12 weeks) were used for preparation of peritoneal exudate of gut derived sepsis by cecal ligation and perforation( CLP).The other 72 rats were randomly divided into portal vein sham group( PV-S),portal vein exudate group( PV-E) and vena cava exudate group( VC-E),24 rats in each group.Normal saline for 1 ml was injected into hepatic portal vein in PV-Sgroup rats.The same amount of peritoneal exudate of gut origin sepsis was injected into rat body in PV-E group and VC-E group through the hepatic portal vein and inferior vena cava.Each group was divided into four subgroups according to intraperitoneal injection at 1,2,4 and 6 h.The abdominal aorta blood of subgroup was collected on different time points.The rats were executed and bronchoalveolar lavage fluid( BALF) was collected to detect the volume of endotoxin,tumor necrosis factor alpha( TNF-α) and interleukin-6( IL-6) in serum and BALF,calculate the wet/dry weight ratio( W/D) and observe morphological and pathological changes of the lung.Results Concentrations of endotoxin,TNF-α,and IL-6and lung pathology score and lung W/D of PV-E group rat were significantly higher than those of PV-S groupafter peritoneal exudate injection 1,2,4,6 h( P < 0.05).Compared to PV-E group,the levels of endotoxin in VC-E group rats were increased and TNF-αand IL-6 in serum and BALF were decreased after peritoneal exudate injection 1,2,4,6 h( P < 0.05).Compared with PV-S group,the rats showed pulmonary edema,extensive and patchy hemorrhage and congestion in PV-E group.The W/D in PV-E group( 1 h:8.53±2.05,2 h:9.41±1.24,4 h:10.35±0.69,6 h:11.05±1.95) was higher than that of PV-S group( 1 h:5.16±1.13,2 h:5.17±1.34,4 h:5.14±2.04,6 h:5.13±1.95) and VC-E group( 1 h:6.71±0.73,2 h:7.65±1.32,4 h:8.40±0.43,6 h:9.41±1.87,P < 0.05).The degree of lung injury in group VC-E was lighter than that of PV-E group.Pulmonary histology showed a large number
出处
《中华卫生应急电子杂志》
2017年第1期48-56,共9页
Chinese Journal of Hygiene Rescue(Electronic Edition)
基金
国家自然科学基金(81673801
81473512
81202831
81173397)
