中国中西医结合急救杂志
中國中西醫結閤急救雜誌
중국중서의결합급구잡지
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care
2015年
6期
601-605
,共5页
低分子肝素%地塞米松%大鼠%急性呼吸窘迫综合征%凝血%纤溶%油酸%内毒素
低分子肝素%地塞米鬆%大鼠%急性呼吸窘迫綜閤徵%凝血%纖溶%油痠%內毒素
저분자간소%지새미송%대서%급성호흡군박종합정%응혈%섬용%유산%내독소
Low molecular weight heparin%Dexamethasone%Rat%Acute respiratory distress syndrome%Coagulation%Fibrinolysis%Oleic acid%Lipopolysaccharide
目的：探讨早期低分子肝素联合地塞米松对油酸及脂多糖（OA-LPS）双重打击诱导的急性呼吸窘迫综合征（ARDS）大鼠凝血/纤溶的影响。方法将40只成年健康雄性SD大鼠按随机数字表法分为假手术组、ARDS模型组、地塞米松组、低分子肝素组和联合治疗组，每组8只。采用经尾静脉注射OA 0.2 mL/kg联合腹腔注射LPS 5 mg/kg序贯双重打击制备大鼠ARDS模型。地塞米松组制模后腹腔注射地塞米松10 mg/kg，低分子肝素组静脉注射低分子肝素200 U/kg，联合治疗组制模后腹腔注射地塞米松10 mg/kg并静脉注射低分子肝素200 U/kg；假手术组经尾静脉注射生理盐水（NS）0.4 mL/kg及腹腔注射NS 2 mL/kg，4 h后腹腔注射NS 1 mL/kg，其余同模型组。以制模后6 h作为实验终点。光镜下观察肺组织病理改变；测定大鼠动脉血氧分压（PaO2）并计算氧合指数（PaO2/FiO2）；测定肺组织湿/干质量（W/D）比值；采用凝固法检测大鼠凝血指标；酶联免疫吸附试验（ELISA）测定血清纤溶酶原激活物抑制剂-1（PAI-1）和Ⅲ型前胶原（PC-Ⅲ）含量。结果光镜下模型组大鼠可见肺间质和肺泡内有红细胞渗出及纤维素沉积，肺泡壁有透明膜形成，肺间质有炎性细胞浸润，肺毛细血管或肺小动脉内可见纤维素性血栓形成；地塞米松组、低分子肝素组和联合治疗组肺间质和肺泡内红细胞渗出及纤维素沉积减少，肺间质炎性细胞浸润减轻，肺毛细血管或肺小动脉内纤维素性血栓减少，以联合治疗组改善最明显。与假手术组比较，模型组PaO2/FiO2明显降低〔mmHg（1 mmHg＝0.133 kPa）：272.02±28.28比420.24±35.52，P＜0.01〕，肺W/D比值显著升高（5.59±0.40比3.82±0.28，P＜0.01），凝血酶原时间（PT）、凝血酶时间（TT）明显延长〔PT（s）：18.78±1.57比16.36±0.97，TT（s）：39.02±5.03比29.22±8.83，均P＜0.05〕，纤维蛋白原（Fib）含量明显降低（g/L：1.82±0.26比2.69±0.40，P＜0.01），血清PAI-1、PC-Ⅲ含量明显升高〔PAI-1（ng/L）：719.04±103.74比517.25±119.18，PC-Ⅲ（μg/L）：29.93±3.24比22.97±6.26，均P＜0.01〕。与模型组比较，地塞米松组、低分子肝素组及联合治疗组3组PaO2/FiO2明显升高，肺W/D比值明显降低，以联合治疗组变化最为明显〔PaO2/FiO2（mmHg）：376.78±25.25比272.02±28.28，肺W/D比值：4.14±0.42比5.59±0.40，均P＜0.01〕，低分子肝素组TT延长最为显著（s：52.00±4.24比39.02±5.03，P＜0.05），3组Fib含量均明显增加（g/L：2.37±0.38、2.59±0.51、2.59±0.24比1.82±0.26，P＜0.05或P＜0.01），而联合治疗组PAI-1含量降低最为显著（ng/L：546.02±93.94比719.04±103.74，P＜0.01）。联合治疗组除PT较地塞米松组及低分子肝素组明显延长（s：19.98±1.61比17.20±1.48、17.02±2.34，均P＜0.05）外，其余指标3组间差异均无统计学意义。结论由OA-LPS双重打击诱导的ARDS大鼠存在凝血功能障碍和纤溶抑制；早期低分子肝素治疗可改善ARDS大鼠凝血及纤溶状况，且联合地塞米松的效果优于单独用药。
目的：探討早期低分子肝素聯閤地塞米鬆對油痠及脂多糖（OA-LPS）雙重打擊誘導的急性呼吸窘迫綜閤徵（ARDS）大鼠凝血/纖溶的影響。方法將40隻成年健康雄性SD大鼠按隨機數字錶法分為假手術組、ARDS模型組、地塞米鬆組、低分子肝素組和聯閤治療組，每組8隻。採用經尾靜脈註射OA 0.2 mL/kg聯閤腹腔註射LPS 5 mg/kg序貫雙重打擊製備大鼠ARDS模型。地塞米鬆組製模後腹腔註射地塞米鬆10 mg/kg，低分子肝素組靜脈註射低分子肝素200 U/kg，聯閤治療組製模後腹腔註射地塞米鬆10 mg/kg併靜脈註射低分子肝素200 U/kg；假手術組經尾靜脈註射生理鹽水（NS）0.4 mL/kg及腹腔註射NS 2 mL/kg，4 h後腹腔註射NS 1 mL/kg，其餘同模型組。以製模後6 h作為實驗終點。光鏡下觀察肺組織病理改變；測定大鼠動脈血氧分壓（PaO2）併計算氧閤指數（PaO2/FiO2）；測定肺組織濕/榦質量（W/D）比值；採用凝固法檢測大鼠凝血指標；酶聯免疫吸附試驗（ELISA）測定血清纖溶酶原激活物抑製劑-1（PAI-1）和Ⅲ型前膠原（PC-Ⅲ）含量。結果光鏡下模型組大鼠可見肺間質和肺泡內有紅細胞滲齣及纖維素沉積，肺泡壁有透明膜形成，肺間質有炎性細胞浸潤，肺毛細血管或肺小動脈內可見纖維素性血栓形成；地塞米鬆組、低分子肝素組和聯閤治療組肺間質和肺泡內紅細胞滲齣及纖維素沉積減少，肺間質炎性細胞浸潤減輕，肺毛細血管或肺小動脈內纖維素性血栓減少，以聯閤治療組改善最明顯。與假手術組比較，模型組PaO2/FiO2明顯降低〔mmHg（1 mmHg＝0.133 kPa）：272.02±28.28比420.24±35.52，P＜0.01〕，肺W/D比值顯著升高（5.59±0.40比3.82±0.28，P＜0.01），凝血酶原時間（PT）、凝血酶時間（TT）明顯延長〔PT（s）：18.78±1.57比16.36±0.97，TT（s）：39.02±5.03比29.22±8.83，均P＜0.05〕，纖維蛋白原（Fib）含量明顯降低（g/L：1.82±0.26比2.69±0.40，P＜0.01），血清PAI-1、PC-Ⅲ含量明顯升高〔PAI-1（ng/L）：719.04±103.74比517.25±119.18，PC-Ⅲ（μg/L）：29.93±3.24比22.97±6.26，均P＜0.01〕。與模型組比較，地塞米鬆組、低分子肝素組及聯閤治療組3組PaO2/FiO2明顯升高，肺W/D比值明顯降低，以聯閤治療組變化最為明顯〔PaO2/FiO2（mmHg）：376.78±25.25比272.02±28.28，肺W/D比值：4.14±0.42比5.59±0.40，均P＜0.01〕，低分子肝素組TT延長最為顯著（s：52.00±4.24比39.02±5.03，P＜0.05），3組Fib含量均明顯增加（g/L：2.37±0.38、2.59±0.51、2.59±0.24比1.82±0.26，P＜0.05或P＜0.01），而聯閤治療組PAI-1含量降低最為顯著（ng/L：546.02±93.94比719.04±103.74，P＜0.01）。聯閤治療組除PT較地塞米鬆組及低分子肝素組明顯延長（s：19.98±1.61比17.20±1.48、17.02±2.34，均P＜0.05）外，其餘指標3組間差異均無統計學意義。結論由OA-LPS雙重打擊誘導的ARDS大鼠存在凝血功能障礙和纖溶抑製；早期低分子肝素治療可改善ARDS大鼠凝血及纖溶狀況，且聯閤地塞米鬆的效果優于單獨用藥。
목적：탐토조기저분자간소연합지새미송대유산급지다당（OA-LPS）쌍중타격유도적급성호흡군박종합정（ARDS）대서응혈/섬용적영향。방법장40지성년건강웅성SD대서안수궤수자표법분위가수술조、ARDS모형조、지새미송조、저분자간소조화연합치료조，매조8지。채용경미정맥주사OA 0.2 mL/kg연합복강주사LPS 5 mg/kg서관쌍중타격제비대서ARDS모형。지새미송조제모후복강주사지새미송10 mg/kg，저분자간소조정맥주사저분자간소200 U/kg，연합치료조제모후복강주사지새미송10 mg/kg병정맥주사저분자간소200 U/kg；가수술조경미정맥주사생리염수（NS）0.4 mL/kg급복강주사NS 2 mL/kg，4 h후복강주사NS 1 mL/kg，기여동모형조。이제모후6 h작위실험종점。광경하관찰폐조직병리개변；측정대서동맥혈양분압（PaO2）병계산양합지수（PaO2/FiO2）；측정폐조직습/간질량（W/D）비치；채용응고법검측대서응혈지표；매련면역흡부시험（ELISA）측정혈청섬용매원격활물억제제-1（PAI-1）화Ⅲ형전효원（PC-Ⅲ）함량。결과광경하모형조대서가견폐간질화폐포내유홍세포삼출급섬유소침적，폐포벽유투명막형성，폐간질유염성세포침윤，폐모세혈관혹폐소동맥내가견섬유소성혈전형성；지새미송조、저분자간소조화연합치료조폐간질화폐포내홍세포삼출급섬유소침적감소，폐간질염성세포침윤감경，폐모세혈관혹폐소동맥내섬유소성혈전감소，이연합치료조개선최명현。여가수술조비교，모형조PaO2/FiO2명현강저〔mmHg（1 mmHg＝0.133 kPa）：272.02±28.28비420.24±35.52，P＜0.01〕，폐W/D비치현저승고（5.59±0.40비3.82±0.28，P＜0.01），응혈매원시간（PT）、응혈매시간（TT）명현연장〔PT（s）：18.78±1.57비16.36±0.97，TT（s）：39.02±5.03비29.22±8.83，균P＜0.05〕，섬유단백원（Fib）함량명현강저（g/L：1.82±0.26비2.69±0.40，P＜0.01），혈청PAI-1、PC-Ⅲ함량명현승고〔PAI-1（ng/L）：719.04±103.74비517.25±119.18，PC-Ⅲ（μg/L）：29.93±3.24비22.97±6.26，균P＜0.01〕。여모형조비교，지새미송조、저분자간소조급연합치료조3조PaO2/FiO2명현승고，폐W/D비치명현강저，이연합치료조변화최위명현〔PaO2/FiO2（mmHg）：376.78±25.25비272.02±28.28，폐W/D비치：4.14±0.42비5.59±0.40，균P＜0.01〕，저분자간소조TT연장최위현저（s：52.00±4.24비39.02±5.03，P＜0.05），3조Fib함량균명현증가（g/L：2.37±0.38、2.59±0.51、2.59±0.24비1.82±0.26，P＜0.05혹P＜0.01），이연합치료조PAI-1함량강저최위현저（ng/L：546.02±93.94비719.04±103.74，P＜0.01）。연합치료조제PT교지새미송조급저분자간소조명현연장（s：19.98±1.61비17.20±1.48、17.02±2.34，균P＜0.05）외，기여지표3조간차이균무통계학의의。결론유OA-LPS쌍중타격유도적ARDS대서존재응혈공능장애화섬용억제；조기저분자간소치료가개선ARDS대서응혈급섬용상황，차연합지새미송적효과우우단독용약。
Objective To observe the effects of low molecular weight heparin (LMWH) combined with dexamethasone (Dex) on coagulation and fibrinolysis at the early stage in rats with acute respiratory distress syndrome (ARDS) induced by two-hit of oleic acid (OA) and lipopolysaccharide (LPS).Methods Forty healthy adult male Sprague-Dawley (SD) rats were randomly divided into sham operation, ARDS model, Dex, LMWH and combining therapy groups (8 rats in each group). The rat ARDS model was established by sequential two-hit with intravenous injection of OA 0.2 mL/kg in a tail vein and intra-peritoneal injection of LPS 5 mg/kg. After model establishment, the rats in Dex group were intra-peritoneally injected with 10 mg/kg Dex, and those in LMWH group were intravenously injected with 200 U/kg LMWH, while the rats in combining therapy group were given Dex and LMWH simultaneously with the same dosages and methods as above mentioned respectively. In sham operation group, however, the rats were intravenously injected with 0.4 mL of normal saline (NS) and were given 2 mL/kg of intra-peritoneal NS injection, and they accepted another 1 mL/kg NS intra-peritoneal injection 4 hours later, the other procedures being the same as those in the model group. The experiment was ended at 6 hours after the establishment of ARDS model. A light microscope was used to observe the pathological changes in lung tissues, partial pressure of arterial blood oxygen (PaO2) was measured, and the oxygenation index (PaO2/FiO2) was calculated. Wet to dry weight (W/D) ratio of lung tissues was also checked. Coagulation indexes were measured by solidification method, and the serum level of plasminogen activator inhibitor-1 (PAI-1) as well as the content of procollagen type Ⅲ (PC-Ⅲ) was determined by enzyme linked immunosorbent assay (ELISA).Results Under the light microscope, effusion of red blood cells, fibrin deposit in the lung interstitium and alveoli, formation of transparent membrane at alveolar wall, inflammatory cells infiltration in pulmonary interstitial tissue, and fibrinous thrombi in lung capillaries or lung arterioles were seen in the model group. Compared with model group, the red blood cells effusion and fibrin deposition in the lung interstitium and alveoli were less, inflammatory cells infiltration in pulmonary interstitium was alleviated and the fibrinous blood emboli in the lung capillaries or lung small arterioles were also decreased in Dex, LMWH and combining therapy groups, among the three groups, the best results being in the combining therapy group. Compared with sham operation group, the PaO2/FiO2 was significantly lowered [mmHg (1 mmHg = 0.133 kPa): 272.02±28.28 vs. 420.24±35.52,P < 0.01], the lung W/D ratio was obviously higher (5.59±0.40 vs. 3.82±0.28,P < 0.01), prothrombin time (PT) as well as thrombin time (TT) were markedly longer [PT (s): 18.78±1.57 vs. 16.36±0.97, TT (s): 39.02±5.03 vs. 29.22±8.83, bothP < 0.05], fibrinogen (Fib) content was significantly decreased (g/L: 1.82±0.26 vs. 2.69±0.40,P < 0.01), but both the serum contents of PAI-1 and PC-Ⅲ were remarkably elevated in model rats [PAI-1 (ng/L): 719.04±103.74 vs. 517.25±119.18, PC-Ⅲ (μg/L): 29.93±3.24 vs. 22.97±6.26, bothP < 0.01); Compared with model group, the level of PaO2/FiO2 was significantly elevated, and the lung W/D ratio was obviously decreased in Dex, LMWH and combining therapy groups respectively, the most significant changes being in combining therapy group [PaO2/FiO2 (mmHg): 376.78±25.25 vs. 272.02±28.28, lung W/D ratio: 4.14±0.42 vs. 5.59±0.4, bothP < 0.01] , in LMWH group, the prolongation of TT was the longest (s: 52.00±4.24 vs. 39.02±5.03,P < 0.05), while Fib contents in the three treatment groups were all obviously increased (g/L: 2.37±0.38, 2.59±0.51, 2.59±0.24 vs. 1.82±0.26,P < 0.05 orP < 0.01); meanwhile, the decrease of PAI-1 in combining therapy group was the greatest (ng/L: 546.02±93.94 vs. 719.04±103.74,P < 0.01). The indexes showed no statistically significant differences among the three treatment groups, except that PT in combining therapy group which was significantly longer than that in Dex and LMWH groups (s: 19.98±1.61 vs. 17.20±1.48, 17.02±2.34, bothP < 0.05). Conclusions The rats with ARDS induced by two-hit of OA combined with LPS have coagulation dysfunction and fibrinolytic inhibition. Using LMWH early can improve coagulation and fibrinolytic status in the rats with ARDS, and the therapeutic effects of LMWH plus Dex for treatment of ARDS are better than those of using each of them alone.
