目的 探讨蛇床子素对小鼠肠缺血再灌注(I/R)损伤的保护作用及其机制.方法 SD雄性小鼠按随机数字表随机分为假手术组、I/R组和蛇床子素组,每组15只.假手术组仅开腹;I/R组通过夹闭小鼠肠系膜上动脉60 min,随后再灌注120 min,建立小鼠I/R模型;蛇床子素组在I/R的瞬间尾静脉注射蛇床子素.检测各组小鼠肠组织含水量,肠组织中伊文思蓝含量,比色法测定血清中的超氧化物歧化酶(SOD)和丙二醛水平,全自动生化仪检测血清中二胺氧化酶(DAO)含量,酶联免疫吸附试验(ELISA)检测血清中肿瘤坏死因子(TNF)-α,白细胞介素(IL)-1β,IL-2等炎性因子的水平.组间比较采用单因素方差分析.结果 I/R组的肠组织含水量、伊文思蓝含量和Chui's评分均高于假手术组和蛇床子素组[0.80%±0.03%比0.77%±0.02%和0.78%±0.02%、(0.11±0.04)比(0.05 ±0.02)和(0.06±0.02) μg/mg、3.42±0.73比0.87±0.35和2.63 ±0.58,P<0.05或P<0.01].I/R组血清中DAO、丙二醛、IL-1β和TNF-α水平均明显高于假手术组和蛇床子素组[(18.9±4.0)比(14.5±2.3)和(16.0±2.6) U/L、(8.4±1.2)比(6.9±1.7)和(6.1±2.4)μmol/L、(93±6)比(51±4)和(67±5) ng/L、(467±31)比(235 ±21)和(323±30) ng/L,P<0.01或P<0.05].I/R组血清中SOD活性和IL-2水平均明显低于假手术组和蛇床子素组[(75±7)比(93±16)和(89±5)U/ml、(95±l6)比(198±14)和(139±11)ng/L,均P<0.05].后两组之间比较,各指标差异均无统计学意义(均P >0.05).结论 蛇床子素可以保护I/R损伤小鼠的肠道组织,其机制可能与抗氧化应激和炎症反应有关.
Objective To explore the protective role of osthole in intestinal ischemia-reperfusion (I/R) injury in mice and examine its underlying mechanism.Methods A murine model of intestinal I/R injury was established with clamping of superior mesenteric artery for 120 min and then clamping was relieved for 60 min.Forty-five SD male mice weighing 27-31 g were randomly divided into 3 groups (n =15 each):sham group (S),I/R injury group (I/R) and L/R plus osthole treatment group (Ost +).Intestinal wet/dry weight ratio,superoxide dismutase (SOD),malondialdehyde (MDA) in serum were examined by colourimetric assay and diamine oxidase (DAO) was examined by automatic biochemical analyzer,the levels of tumor necrosis factor (TNF)-α,interleukin (IL)-1β and IL-2 were examined by enzyme-linked immunosorbent assay (ELISA).Intestinal barrier permeability was detected by Evans blue (EB) test.One-way ANOVA was used to analyze all experimental data variance.Results Intestinal tissues wet/dry weight ratios,Evans blue content and Chiu's score of I/R group mice were significantly higher than those of S and Ost+ groups (0.80% ±0.03% vs 0.77% ±0.02% & 0.78% ±0.02%,(0.11 ±0.04)vs (0.05 ±0.02)& (0.06 ±0.02) μg/mg,3.42 ±0.73 vs 0.87 ±0.35 & 2.63 ±0.58,P <0.05 or P < 0.01).Serum level of DAO,MDA,IL-1β & TNF-α of I/R group mice were significantly higher than those of S and Ost+ groups ((18.9±4.0)vs (14.5±2.3)& (16.0±2.6)U/L,(8.4±1.2)vs (6.9±1.7)& (6.1 ±2.4)μmol/L,(93 ±6) vs (51 ±4)& (67 ±5)ng/L,(467±31)vs (235 ±21)& (323 ±30) ng/L,P < 0.01 or P < 0.05).Serum SOD activity and IL-2 level were significantly lower than those of S and Ost+ groups ((7.5 ±7)vs (93 ±16)& (89 ±5)U/ml,(95 ±16)vs (198 ±14)& (139 ± ll)ng/L,all P < 0.05).All parameters showed no significant difference between S and Ost + groups (all P > 0.05).Conclusions Treatment of osthle may protect murine intestinal tissue against intestinal I/R injury.And the mechanisms may be due to its actions of preventing oxygen stress and inflammatory responses.