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
615-618
,共4页
郝雪景%蔡国龙%胡才宝%颜黙磊%饶群
郝雪景%蔡國龍%鬍纔寶%顏黙磊%饒群
학설경%채국룡%호재보%안묵뢰%요군
脓毒症%乌司他丁%内皮细胞%黏附分子%大鼠
膿毒癥%烏司他丁%內皮細胞%黏附分子%大鼠
농독증%오사타정%내피세포%점부분자%대서
Sepsis%Ulinastatin%Endothelial cell%Adhesion molecule%Rat
目的：探讨乌司他丁（UTI）对脓毒症大鼠内皮细胞的保护作用及其机制。方法52只雄性SD大鼠，按随机数字表法分为生理盐水预处理组（对照组）和UTI预处理组（UTI组）两组，每组26只。两组均腹腔注射脂多糖（LPS）10 mg/kg制备脓毒症大鼠模型； UTI组于LPS注射前18 h腹腔注射UTI 100 kU/kg（溶于5 mL生理盐水中）预处理，对照组于LPS注射前3 h腹腔注射5 mL生理盐水预处理。分别于制模后0.5、2、4、12、24、72 h取尾静脉血和心肌组织，采用酶联免疫吸附试验（ELISA）检测血清肿瘤坏死因子-α（TNF-α）、白细胞介素（IL-6、IL-10）和血管细胞黏附分子（VCAM）、细胞间黏附分子-1（ICAM-1）水平，并分析TNF-α与ICAM-1的相关性；用免疫组化法检测心肌细胞ICAM-1表达。结果制模后两组脓毒症大鼠血清TNF-α、IL-6、IL-10、ICAM、VCAM均逐渐升高，分别于24 h、12 h、12 h、72 h、72 h达到峰值。与对照组比较， UTI组各时间点TNF-α、IL-6、ICAM-1、VCAM均明显降低〔24 h TNF-α（ng/L）：119.8±28.9比190.2±30.4，12 h IL-6（ng/L）：327.8±26.9比948.7±63.8，72 h VCAM（ng/L）：36.3±3.2比68.8±2.4，72 h ICAM（ng/L）：115.6±11.6比129.4±8.2，P＜0.05或P＜0.01〕，IL-10明显升高〔12 h（ng/L）：80.7±1.9比42.3±4.9，P＜0.01〕。TNF-α与ICAM呈显著正相关（UTI组：r＝0.907，P＝0.050；对照组：r＝0.961，P＝0.010）。免疫组化显示，两组制模后0.5 h心肌细胞基本无ICAM-1阳性表达；12 h时对照组ICAM-1阳性表达增加，UTI组有少量表达；72 h时两组ICAM-1表达均明显增加。结论 UTI通过调节促炎因子、抑炎因子及黏附分子的表达，改善微血管通透性从而起到保护内皮细胞功能的作用。
目的：探討烏司他丁（UTI）對膿毒癥大鼠內皮細胞的保護作用及其機製。方法52隻雄性SD大鼠，按隨機數字錶法分為生理鹽水預處理組（對照組）和UTI預處理組（UTI組）兩組，每組26隻。兩組均腹腔註射脂多糖（LPS）10 mg/kg製備膿毒癥大鼠模型； UTI組于LPS註射前18 h腹腔註射UTI 100 kU/kg（溶于5 mL生理鹽水中）預處理，對照組于LPS註射前3 h腹腔註射5 mL生理鹽水預處理。分彆于製模後0.5、2、4、12、24、72 h取尾靜脈血和心肌組織，採用酶聯免疫吸附試驗（ELISA）檢測血清腫瘤壞死因子-α（TNF-α）、白細胞介素（IL-6、IL-10）和血管細胞黏附分子（VCAM）、細胞間黏附分子-1（ICAM-1）水平，併分析TNF-α與ICAM-1的相關性；用免疫組化法檢測心肌細胞ICAM-1錶達。結果製模後兩組膿毒癥大鼠血清TNF-α、IL-6、IL-10、ICAM、VCAM均逐漸升高，分彆于24 h、12 h、12 h、72 h、72 h達到峰值。與對照組比較， UTI組各時間點TNF-α、IL-6、ICAM-1、VCAM均明顯降低〔24 h TNF-α（ng/L）：119.8±28.9比190.2±30.4，12 h IL-6（ng/L）：327.8±26.9比948.7±63.8，72 h VCAM（ng/L）：36.3±3.2比68.8±2.4，72 h ICAM（ng/L）：115.6±11.6比129.4±8.2，P＜0.05或P＜0.01〕，IL-10明顯升高〔12 h（ng/L）：80.7±1.9比42.3±4.9，P＜0.01〕。TNF-α與ICAM呈顯著正相關（UTI組：r＝0.907，P＝0.050；對照組：r＝0.961，P＝0.010）。免疫組化顯示，兩組製模後0.5 h心肌細胞基本無ICAM-1暘性錶達；12 h時對照組ICAM-1暘性錶達增加，UTI組有少量錶達；72 h時兩組ICAM-1錶達均明顯增加。結論 UTI通過調節促炎因子、抑炎因子及黏附分子的錶達，改善微血管通透性從而起到保護內皮細胞功能的作用。
목적：탐토오사타정（UTI）대농독증대서내피세포적보호작용급기궤제。방법52지웅성SD대서，안수궤수자표법분위생리염수예처리조（대조조）화UTI예처리조（UTI조）량조，매조26지。량조균복강주사지다당（LPS）10 mg/kg제비농독증대서모형； UTI조우LPS주사전18 h복강주사UTI 100 kU/kg（용우5 mL생리염수중）예처리，대조조우LPS주사전3 h복강주사5 mL생리염수예처리。분별우제모후0.5、2、4、12、24、72 h취미정맥혈화심기조직，채용매련면역흡부시험（ELISA）검측혈청종류배사인자-α（TNF-α）、백세포개소（IL-6、IL-10）화혈관세포점부분자（VCAM）、세포간점부분자-1（ICAM-1）수평，병분석TNF-α여ICAM-1적상관성；용면역조화법검측심기세포ICAM-1표체。결과제모후량조농독증대서혈청TNF-α、IL-6、IL-10、ICAM、VCAM균축점승고，분별우24 h、12 h、12 h、72 h、72 h체도봉치。여대조조비교， UTI조각시간점TNF-α、IL-6、ICAM-1、VCAM균명현강저〔24 h TNF-α（ng/L）：119.8±28.9비190.2±30.4，12 h IL-6（ng/L）：327.8±26.9비948.7±63.8，72 h VCAM（ng/L）：36.3±3.2비68.8±2.4，72 h ICAM（ng/L）：115.6±11.6비129.4±8.2，P＜0.05혹P＜0.01〕，IL-10명현승고〔12 h（ng/L）：80.7±1.9비42.3±4.9，P＜0.01〕。TNF-α여ICAM정현저정상관（UTI조：r＝0.907，P＝0.050；대조조：r＝0.961，P＝0.010）。면역조화현시，량조제모후0.5 h심기세포기본무ICAM-1양성표체；12 h시대조조ICAM-1양성표체증가，UTI조유소량표체；72 h시량조ICAM-1표체균명현증가。결론 UTI통과조절촉염인자、억염인자급점부분자적표체，개선미혈관통투성종이기도보호내피세포공능적작용。
Objective To approach the effect of ulinastatin (UTI) on protection of vascular endothelial cells in rats with sepsis and its mechanism.Methods Fifty-two Sprague-Dawley (SD) male rats were randomly divided into a normal saline pretreatment group (control group) and a UTI pretreatment group (UTI group), each groupn = 26. The rats in two groups were given lipopolysaccharide (LPS, 10 mg/kg) intra-peritoneal injection for the establishment of rat septic models. In UTI group, 18 hours before LPS injection, intraperitoneal injection of UTI 100 kU/kg dissolved in 5 mL normal saline was given, while in the control group, 3 hours before LPS injection, intraperitoneal injection of 5 mL normal saline was given to the rats for pretreatment. Respectively, at 0.5, 2, 4, 12, 24, 72 hours after model establishment, tail venous blood and myocardial tissue were taken. The levels of tumor necrosis factor-α (TNF-α), interleukins (IL-6, IL-10), vascular cell adhesion molecule (VCAM) and intercellular adhesion molecule-1 (ICAM-1) were detected by enzyme-linked immunosorbent assay (ELISA); the correlation between TNF-α and ICAM-1 was analyzed; the expression of ICAM-1 in myocardial cell was determined by immunohistochemistry.Results After model establishment, the levels of TNF-α, IL-6, IL-10, ICAM and VCAM in two groups were gradually increased, reaching the peaks at 24, 12, 12, 72, 72 hours, respectively. Compared with control group, the levels of TNF-α, IL-6, ICAM-1, VCAM of UTI group were significantly lower at various time points [24 hours TNF-α (ng/L): 119.8±28.9 vs. 190.2±30.4, 12 hours IL-6 (ng/L): 327.8±26.9 vs. 948.7±63.8, 72 hours VCAM (ng/L): 36.3±3.2 vs. 68.8±2.4, 72 hours ICAM-1 (ng/L): 115.6±11.6 vs. 129.4±8.2,P < 0.05 orP < 0.01], IL-10 was significantly increased [12 hours (ng/L): 80.7±1.9 vs. 42.3±4.9,P < 0.01]. TNF-αwas positively correlated to ICAM significantly (UTI group:r = 0.907,P = 0.050; control group:r = 0.961, P = 0.010). Immunohistochemistry showed that after modeling for 0.5 hour, basically no positive expression of ICAM-1 in myocardial cells was found in the two groups; in the control group, at 12 hours the positive expression of ICAM-1 was increased, and in UTI group, a little expression of ICAM-1 was seen; at 72 hours, the expression of ICAM-1 was significantly increased in both groups.Conclusion UTI can protect the function of endothelial cells in rats with sepsis by regulating the expressions of proinflammatory cytokine, anti-inflammatory cytokine, adhesion molecules, and improving the microvascular permeability.
