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淋巴引流及ω-3多不饱和脂肪酸干预对大鼠肠道缺血再灌注的影响

Effects of lymphatic drainage and ω-3 polyunsaturated fatty acids on intestinal ischemia-reperfusion injury in rats

摘要目的 观察大鼠肠道缺血再灌注(Ⅰ/R)损伤时肠淋巴液引流对高迁移率族蛋白1(HMGB1)、炎症因子和内毒素的影响以及ω-3多不饱和脂肪酸(ω-3 PUFA)干预的效果.方法 72只SD大鼠随机区组法随机分为单纯引流组、Ⅰ/R组、Ⅰ/R+引流组(每组8只)和胃造口组[正常饮食(N)组、普通肠内营养(EN)组、普通肠内营养加ω-3 PUFA(PUFA)3大组,每大组再根据是否行Ⅰ/R 和引流分为2组,每组8只].单纯引流组只引流180 mⅠn淋巴液不行Ⅰ/R损伤;Ⅰ/R、VR+引流组行肠系膜上动脉夹闭60 mⅠn再灌注120 mⅠn;Ⅰ/R+引流组同时行肠淋巴液引流180 mⅠn.胃造口组大鼠均先行胃造口手术,分别给予不同营养5 d后造模,各引流组同前进行肠淋巴液引流180 mⅠn.手术完毕后分别取血清和淋巴液,定量检测内毒素,酶联免疫吸附试验(ELⅠSA)定量检测炎症因子以及HMGB1.结果 Ⅰ/R+引流组淋巴液中内毒素、炎症因子以及HMGBl均高于单纯引流组[均P<0.05,白细胞介素(ⅠL)-6(30±8)pg/ml比(20±6)pg/ml,内毒素(0 029±0.011)U/ml比(0 008±0 005)U/ml];Ⅰ/R+引流组血清中内毒素、炎症因子均低于Ⅰ/R组(均P<0 05).在胃造口组中,N 组和EN组的淋巴液中肿瘤坏死因子(TNF)-α与HMGBl均高于PUFA组[(46±17)pg/ml、(54±16)pg/ml比(28±9)pg/ml,(4.8±1.6)ng/ml、(5.3±1.8)ns/ml比(3.0±1.0)ng/ml,均P<0.05].PUFA(Ⅰ/R)组血清中内毒素、炎症因子以及HMGBl均低于N(Ⅰ/R)组(均P<0.05),PUFA(Ⅰ/R+引流)组血清中TNF-α与HMGBl均低于N(Ⅰ/R+引流)组(均P<0 05).结论 引流肠淋巴液能够降低肠道Ⅰ/R损伤时内毒素、炎症因子和HMGB1的水平,减轻大鼠肠道Ⅰ/R引起的损伤.ω-3PUFA的干预对于肠道Ⅰ/R引起的损伤有一定的保护作用,对于减轻炎症反应有积极作用.

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abstractsObjective To investigate the effects of lymphatic drainage and ω-3 polyunsaturated fatty acid (ω-3PUFA) on high mobility group box 1 (HMGB1) , inflammatory cytokines and endotoxin in rats with intestinal ischemia-reperfusion (Ⅰ/R) injury. Methods A total of 72 SD rats were randomly divided into drainage-alone group, Ⅰ/R group, ischemia-reperfusion plus drainage (Ⅰ/R+D) group (n=8 each)and 3 groups with 16 rats undergoing gastrostomy in each group: normal diet (N) group, enteral nutrition (EN) group and enteral nutrition & ω-3PUFA (PUFA) group. And they were further divided into 2 subgroups (n=8). The rats in Ⅰ/R and Ⅰ/R+D groups were subjected to a 60-min ischemia follow by 120-min reperfusion injury of superior mesenteric artery. When the rats suffered Ⅰ/R injury, intestinal lymph was drained for 180 min in the Ⅰ/R+D group. The rats in the drainage-alone group received 180-min lymph drainage without Ⅰ/R injury. After 5 days with different nutrition regimes, the models were established similarly. The rats in the Ⅰ/R+D sub-groups were treated with intestinal lymph drainage for 180 min. The serum and lymph samples were collected post-operatively. Endotoxin was detected by a Limulus kit. The inflammatory cytokines and high mobility group box 1 (HMGB1) were analyzed by enzyme-linked immunosorbent assay (ELISA).Results Endotoxin, inflammatory cytokines and lymphatic HMGB1 of lymphatic in the Ⅰ/R+D group were higher than those in the drainage-alone group [all P<0.05 , IL-6 :(30±8) pg/ml vs (20±6) pg/ml, endotoxin: (0.029±0.011) U/ml vs (0.008+0.005) U/ml].The serum levels of endotoxin and inflammatory cytokines in the Ⅰ/R+ D group were lower than those in the Ⅰ/R group (P<0.05).The lymphatic levels of TNF-a (tumor necrosis factor-alpha) and HMGB1 in the N and EN groups were higher than those in the PUFA group[TNF-α: (46±17)pg/ml, (54±16)pg/ml vs(28±9) pg/ml, HMGB1: (4.8±1.6) ng/ml, (5.3±1.8) ng/ml, (3.0±1.0) ng/ml, all P<0.05) ].The serum levels of endotoxin, inflammatory cytokines and HMGB1 in the PUFA(l/R) group were lower than those in the N(Ⅰ/R) group (F<0.05).The levels of TNF-a and HMGB1 were lower in the PUFA (Ⅰ/R+D) group than those in the N(Ⅰ/R+ D) group (both P<0.05).Conclusion Lymphatic drainage may reduce the levels of endotoxin, inflammatory cytokines and HMGB1 so as to alleviate the intestinal Ⅰ/R injury. The intervention of ω-3PUFA has some protective effect through relieving inflammation.

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中华医学杂志

中华医学杂志

2011年91卷25期

1775-1779页

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