摘要
目的探讨阿托伐他汀（ATO）联合低分子肝素（LMWH）对脓毒症大鼠炎症反应及肺脏的保护作用。方法选择健康雄性SD大鼠122只，按随机数字表法分为假手术（Sham）组（n=10）、脓毒症组（n=10）、ATO组（n=34）、LMWH组（n=34）、ATO联合LMWH组（联合组，n=34）。采用盲肠结扎穿孔术（CLP）制备脓毒症大鼠模型；Sham组只开腹，不结扎、穿孔。各预处理组于术前相应灌胃ATO20mg／kg以及皮下注射LMWH100U／kg或二者联用，均连续给药5d。依据改良脓毒症严重程度评定标准对各组大鼠的病情程度进行评分；每组取10只大鼠观察7d存活情况。于术前（0h）及术后4、8、12、24h取血，采用酶联免疫吸附试验（ELISA）检测血浆肿瘤坏死因子-α（TNF—α）、白细胞介素（IL-1β）、高迁移率族蛋白β1（HMGβ1）水平；术后24h取肺组织，苏木素-伊红（HE）染色后光镜下观察病理学改变。结果①脓毒症组术后4h病情严重程度评分即较Sham组明显增高（分：12．2±2．0比7．2±0．5，P〈0．05），且呈逐渐升高趋势，7d累积死亡率达90％（9／10）。ATO组、LMWH组、联合组8h起疴隋严重程度评分均较脓毒症组明显下降（分：12．2±2．0、11．2±2．2、10．0±1．7比16．6±2．5，均P〈0．05），7d累积死亡率分别为60％（6／10）、60％（6／10）、40％（4／10），均较脓毒症组显著降低（均P〈0．05）.②Sham组术后各炎性因子水平无明显变化；其他4组各炎性因子水平均较术前明显升高，TNF—α在4h、IL-1β在8h、HMGBl在24h达峰值。脓毒症组各炎性因子水平均较Sham组显著升高；而ATO组、LMWH组、联合组各炎性因子水平均明显低于脓毒症组[4hTNF—α（ng／L）：668．3±124．6、536．5±118．5、496．5±108．5比783．8±134．7，8hIL-1β（ng／L）：2476．7±137．8、2460．4±171．2、2090．0±151．2比2873．9±295．6，24hHMGB1（μg/L）：654．4±154．4、659．0±134．6、609．4±90．5比859．3±167．5，P〈0．05或P〈0．01]。③光镜下观察，Sham组肺泡组织形态正常；脓毒症组肺脏病理损害较为严重；而3个预处理组肺脏病理损害较脓毒症组明显减轻，以联合组改善最为明显。结论ATO、LMWH可降低脓毒症大鼠严重程度，抑制血浆早期及晚期炎性因子释放，减轻肺部病理改变，降低死亡率，联合应用ATO和LMWH效果更佳。
Objective To investigate the influence of combined use of atorvastatin （ATO） and low molecular weight heparin （LMWH） on the inflammatory reaction and pulmonary protection functions in rats with sepsis. Methods A total of 122 healthy male Sprague-Dawley （SD） rats were divided into five groups using a random number table： sham-operated group （sham group, n = 10）, sepsis group＇ （n = 10）, ATO group （n=34）, LMWH group （n = 34）, and ATO combined with LMWH group （ATO＋LMWH group, n = 34）. The rat model of sepsis was reproduced by cecal ligation and puncture （CLP）, while in sham group, rats were only subjected to laparotomy without cecum ligation and puncture. The rats of each pretreatment group received relevant therapies for 5 days, either gastric perfusion with ATO 20 mg/kg or subcutaneous injection with LMWH 100 U/kg or both before operation. The sepsis severities of the model animals were scored according to the modified sepsis severity assessment standards of experimental animals. Ten rats in each group were calculated the 7-day cumulative mortality rate. Blood samples from 6 rats in each group were collected to determine the levels of tumor necrosis factor- α （TNF- α）, interleukin-1β （IL-1 β ） and high mobility group protein box-1 （HMGB1） contents in plasma using enzyme linked immunosorbent assay （ELISA） before operation （0 hour） and 4, 8, 12, and 24 hours post operation. The lung tissue was harvested 24 hours after operation, and the pulmonary pathology was assayed by hematoxylin and eosin （HE） staining using optical microscope. Results ① The sepsis severity grades of sepsis group were significantly higher than those of sham group at 4 hours after operation （score： 12.2 ± 2.0 vs. 7.2 ± 0.5, P 〈 0.05 ）. Furthermore, they displayed a gradually increasing tendency, with the 7-day cumulative mortality rate being 90% （9/10）. The sepsis severity grades in ATO group, LMWH group, and ATO＋LMWH group showed a significant decrease compared with sepsis group at 8 hours after operation （12.2 ±2.0, 11.2 ± 2.2, 10.0 ± 1.7 vs. 16.6±2.5, all P 〈 0.05）. The 7-day cumulative mortality rates in ATO group, LMWH group, and ATO＋LMWH group were 60% （6/10）, 60% （6/10）, and 40% （4/10）, respectively, all of which was significantly lower than that of sepsis group （all P 〈 0.05）. ② The levels of TNF- α, IL-1β and HMGB1 have not shown much variations in the sham group after operation; the levels of pro-inflammatory cytokines in other 4 groups were significantly increased after operation compared with those before operation; the levels of TNF- α, IL-1β, and HMGB1 reached peak at 4, 8, and 24 hours, respectively. The levels of pro-inflammatory cytokines in sepsis group were significantly higher than those in the sham group. However, the levels of pro-inflammatory cytokines in ATO group, LMWH group, and ATO＋LMWH group were significantly lower than those in sepsis group [4-hour TNF- α （ng/L）： 668.3 ± 124.6, 536.5 ± 118.5, 496.5± 108.5 vs. 783.8 ±134.7; 8-hour IL-113 （ng/L）： 2 476.7± 137.8, 2 460.4 ± 171.2, 2 090.0 ± 151.2 vs. 2 873.9 ±295.6; 24-hour HMGB1 （μg/L）： 654.4 ± 154.4, 659.0 ± 134.6, 609.4± 90.5 vs. 859.3 ± 167.5, P 〈 0.05 or P 〈 0.01]. ③ It was showed by optical microscopy that the pulmonary tissue morphology was normal in sham group and that the damage of pulmonary pathology was relatively severe in sepsis group. Compared with sepsis group, the damage of pulmonary pathology in ATO group, LMWH group, and ATO ＋ LMWH group was alleviated obviously, and the most obvious improvements were found in ATO ＋ LMWH group. Conclusions Either ATO or LMWH could decrease sepsis severity, suppress the release of plasma pro-inflammatory cytokines at the early and late stages, alleviate the damage of pulmonary pathology, and reduce the 7-day cumulative mortality rate. Therefore, the combined treatment of sepsis using both ATO and LMWH resulted in better outcomes than implemented individually.
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2016年第5期427-432,共6页
Chinese Critical Care Medicine
基金
山东省自然科学基金（ZR2013HM062）
国家自然科学基金青年基金（81200238）
山东省优秀中青年科学家科研奖励基金（BS2011YY043）
关键词
脓毒症
盲肠结扎穿孔术
阿托伐他汀
低分子肝素
炎性因子
Sepsis
Cecal ligation and puncture
Atorvastatin
Low molecular weight heparin
Pro-inflammatory cytokine
作者简介
通讯作者：王春亭，Email：WCTEICU@126．com
