脂多糖致大鼠急性肺损伤模型取材时间选择

Selection of sampling time for preparing lipopolysaccharide-induced acute lung injury rat model

目的探讨制备大鼠急性肺损伤模型过程中取材时间的选择。方法 30只成年雄性SD大鼠,随机分为生理盐水对照组(对照组)6只和脂多糖组24只。脂多糖组大鼠尾静脉注射浓度为10mg/kg脂多糖1mL建立急性肺损伤模型,对照组大鼠尾静脉注射1mL生理盐水。对照组和脂多糖组于注药后1、3、6、12h各取大鼠6只,检测肺组织湿、干质量比,采用光镜观察肺组织病理形态学,采用免疫组织化学法检测肺组织白细胞介素-6和血管内皮生长因子表达情况。结果注射脂多糖1h后,脂多糖组光镜下可见肺间质水肿和炎症细胞浸润;脂多糖组1、3、6、12h时肺组织湿/干质量比(5.57±0.55、5.81±0.81、6.11±0.68、5.84±0.52)、肺损伤评分((1.17±0.41)、(2.33±0.52)、(3.50±0.55)、(2.83±0.75)分)、肺组织白细胞介素-6((31.33±6.95)、(78.33±7.53)、(95.17±10.93)、(72.00±10.30)ng/L)和血管内皮生长因子((228.17±17.30)、(284.33±18.74)、(384.50±29.22)、(316.33±26.80)ng/L)水平明显高于对照组((5.19±0.98)、(0.00±0.00)分、(9.00±3.85)ng/L、(197.50±15.18)ng/L)(P<0.05),其中6h时明显高于1、3、12h时;3、12h时高于1h时(P<0.05)。结论尾静脉注射10mg/kg脂多糖在6h时可成功复制大鼠急性肺损伤模型。

Objective To explore the selection of sampling time during the process for preparing lipopolysaccharide（LPS）-induced acute lung injury rat model.Methods Thirty adult male SD rats were randomly divided into control group（n=6）and LPS group（n=24）.LPS group was injected 1mL LPS at 10mg/kg concentration via caudal vein to establish acute lung injury model,while control group was injected l mL normal saline via caudal vein.Six rats were collected from control group and LPS group after 1,3,6and 12 hours of injection,to detect the wet to dry ratio of lung tissue.The pathology morphology was observed by light microscope,and expressions of interleukin-6and vascular endothelial growth factor（VEGF）in lung tissue were detected by immunohistochemistry method.Results One hour after LPS injection,light microscope showed lung interstitial edema and inflammatory cell infiltration in LPS group.After 1,3,6and 12 hours,the wet to dry ratios of lung tissue were 5.57±0.55,5.81±0.81,6.11±0.68 and 5.84±0.52,lung injury rating scores were 1.17±0.41,2.33±0.52,3.50±0.55 and 2.83±0.75,interleukin-6levels in lung tissue were（31.33±6.95）,（78.33±7.53）,（95.17±10.93）and（72.00±10.30）ng/L,and VEGF levels were（228.17±17.30）,（284.33±18.74）,（384.50±29.22）and（316.33±26.80）ng/L in LPS group,significantly higher than those in control group（5.19±0.98,0.00±0.00,（9.00±3.85）ng/L,（197.50±15.18）ng/L）（P〈0.05）,among which,the data after 6hours were the highest,and the data after 3and 12 hours were higher than those after 1hour（P〈0.05）.Conclusion The acute lung injury rat model can be successfully established 6hours after injection of 10mg/kg LPS via caudal vein.