摘要
目的 探讨微小RNA （miR）-155对脂多糖（LPS）所致小鼠急性肺损伤的干预作用.方法 将30只小鼠按照随机数字表法分为LPS致伤组、miR-155抑制组、miR对照组、miR-155增强组、空白对照组5组各6只.LPS致伤组和其他3个处理组麻醉后行气管穿刺给予5μl LPS（浓度为20 μg/μl,4 mg/kg）建立小鼠急性肺损伤模型,致伤后30 min起,LPS致伤组气管给予生理盐水;miR-155抑制组、miR对照组、miR-155增强组分别气管给予miR-155抑制剂,miR对照剂,miR-155增强剂,空白对照组不做任何处理.24 h后收集各组肺组织：HE染色观察组织形态,绿色荧光染色法检测各组肿瘤坏死因子-α（TNF-α）,白细胞介素-1β （IL-1β）的mRNA表达水平,Taqman法检测miR-155抑制组、miR对照组、miR-155增强组中miR-155的表达含量,Western印迹法检测各组环氧合酶-2（COX-2）蛋白的表达,并对数据进行统计学分析.结果 LPS致伤组、miR-155抑制组、miR对照组、miR-155增强组、空白对照组的TNF-α mRNA相对表达量分别为24.49±3.64、38.92±3.40、36.00±3.86、12.56±2.06、1.04 ±0.41;IL-1β mRNA相对表达量分别为55.96±4.87、62.90±6.41、59.99±6.48、25.33±3.50、1.11±0.44,miR-155增强组与LPS致伤组、miR-155抑制组、miR对照组差异均有统计学意义（均P＜ 0.05）,而LPS致伤组、miR-155抑制组、miR对照组间差异均无统计学意义;上述5组COX-2蛋白相对表达量的灰度值分别为0.94±0.06、0.96±0.09、0.91 ±0.03、0.79±0.04、0.63±0.07,空白对照组均显著低于其他各组（均P＜0.05）.结论 miR-155可有效减弱LPS所致小鼠急性肺损伤炎症反应.
Objective To study the effects of microRNA （miR）-155 on mice with acute lung injury induced by lipopolysaccharide （LPS）.Methods According to the random number table,30 C57 mice were divided into group LPS,group miR-155 inhibitor,group miR control,group miR-155 mimics,normal control group,with 6 mice in each group.Mice in group LPS and the other three treatment groups were given 5 μl LPS （20 μg/μl,in the dose of 4 mg/kg） to reproduce acute lung injury model.Half an hour after injury,group LPS,group miR-155 inhibitor,group miR control,group miR-155 mimics were injected with saline,miR-155 inhibitor,miR control and miR-155 mimics through trachea.Mice in normal control group were received no treatment.All mice were anesthetized after 24 h,while lung tissue samples were harvested for histomorphological observation,detection of mRNA expressions of tumor necrosis factor-α （TNF-α）,interleukin-1 β （IL-1 β） with SYBR Green real-time polymerase chain reaction （PCR）,detection of miR-155 in group miR-155 inhibitor,group miR control,group miR-155 mimics by Taqman PCR.The protein expression of cyclooxygenase-2 （COX-2） was determined by western blot.Data were statistically analyzed.Results The relative mRNA expression values of TNF-α were 24.49 ± 3.64,38.92 ± 3.40,36.00 ±3.86,12.56 ± 2.06,1.04 ± 0.41 in group LPS,group miR-155 inhibitor,group miR control,group miR-155 mimics,normal control group.The relative mRNA expression values of IL-1 β were 55.96 ± 4.87,62.90 ± 6.41,59.99 ± 6.48,25.33 ± 3.50,1.11 ± 0.44 in group LPS,group miR-155 inhibitor,group miR control,group miR-155 mimics,normal control group.The differences of TNF-α and IL-1β in group miR-155 mimics were statistically significant by compared with group LPS,group miR-155 inhibitor,group miR control （P ＜ 0.05）.The differences were not statistically significant among group LPS,group miR-155 inhibitor,group miR control.Western blot analysis showed the relative expression values of COX-2 were 0.94±0.06,0.96 ±0.09,0.91 ±0.03,0.79 ±0.04,0.63 ±0.07 in group LPS,group miR-155 inhibitor,group miR control,miR-155 mimics and normal control group.The value in normal control group was significantly lower than other groups （P ＜ 0.05）.Conclusion Using the miR-155 can effectively alleviate the lung inflammation in mice with acute lung injury induced by LPS.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2015年第28期2312-2316,共5页
National Medical Journal of China
基金
国家自然科学基金（81372051）
总参军事医学和老年病科研基金（ZCWS14B06）
