摘要
目的探讨miR-34b-5p对脂多糖(LPS)诱导的急性呼吸窘迫综合征(ARDS)大鼠的肺部炎症、细胞凋亡的影响及可能机制。方法 60只SD大鼠分为对照组、ARDS、ARDS+miR-34b-5p mimics mock及ARDS+miR-34b-5p mimics组,每组15只。除对照组外,其余组大鼠腹腔注射LPS 3 mg/mL,ARDS+miR-34b-5p mimics mock和ARDS+miR-34b-5p mimics组建模后分别尾静脉注射50μL miR-34b-5p mimics mock和miR-34b-5p mimics,对照组大鼠腹腔和尾静脉注射等量生理盐水。RT-PCR检测肺组织中miR-34b-5p mRNA表达,苏木素-伊红(HE)染色观察大鼠肺组织病理变化,计算大鼠肺组织W/D值,TUNEL法检测细胞凋亡率,ELISA法检测白介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)水平,Western blot检测颗粒体蛋白前体(PGRN)蛋白表达,生物信息预测miR-34b-5p与PGRN的靶向作用关系,荧光素酶实验验证。结果 ARDS组大鼠肺组织中miR-34b-5p mRNA水平低于对照组,差异有统计学意义(P<0.01);ARDS+miR-34b-5p mimics组大鼠肺组织中miR-34b-5p mRNA水平高于ARDS组,差异有统计学意义(P<0.01);ARDS+miR-34b-5p mimics mock组大鼠肺组织中miR-34b-5p mRNA水平高于ARDS组,差异无统计学意义(P>0.05)。对照组大鼠肺组织规则,形态完整,清晰;ARDS组大鼠肺组织结构紊乱,肺泡内可见大量的红细胞及炎性细胞浸润。与ARDS组比较,ARDS+miR-34b-5p mimics mock组大鼠肺组织病理改变无明显差别,ARDS+miR-34b-5p mimics组大鼠肺组织较规则,红细胞、炎性细胞少。ARDS组大鼠肺组织W/D值、细胞凋亡率高于对照组,ARDS+miR-34b-5p mimics组大鼠肺组织W/D值、细胞凋亡率低于ARDS组,差异均有统计学意义(P<0.05);ARDS组大鼠外周血IL-6和TNF-α水平高于对照组,ARDS+miR-34b-5p mimics组大鼠外周血IL-6和TNF-α水平低于ARDS组,差异均有统计学意义(P<0.05);ARDS组大鼠肺组织中PGRN蛋白水平低于对照组,ARDS+miR-34b-5p mimics组大鼠组织中PGRN蛋白水平高于ARDS组,差异均有统计学意义(P<0.05)。miR-34b-5p mi
Objective To investigate the effect of miR-34 b-5 p on pulmonary inflammation and cell apoptosis of lipopolysaccharide(LPS)-induced acute respiratory distress syndrome(ARDS)rats and its possible mechanism. Methods60 rats were divided into control group,ARDS group,ARDS+miR-34 b-5 p mimics mock group and ARDS+miR-34 b-5 p mimics group,15 rats in each group. Except for the control group, rats in the other groups were intraperitoneally injected with LPS 3 mg/mL. Rats in ARDS + miR-34 b-5 p mimics mock and ARDS + miR-34 b-5 p mimics groups were intraperitoneally injected 50 μL miR-34 b-5 p mimics mock and miR-34 b-5 p mimics. After modeling, rats in the control group were intraperitoneally injected with the same amount of normal saline. RT-PCR was used to detect the expression of miR-34 b-5 p mRNA in lung tissue. HE staining was used to observe the pathological changes of lung tissue. W/D value of lung tissue was calculated. TUNEL was used to detect apoptosis ratio. ELISA method was used to detect interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α). The expression of progranulin(PGRN) protein was detected by Western blot. Biological information was used to predict the targeting effect of miR-34 b-5 p and PGRN. Luciferase test was used to verify. Results The level of miR-34 b-5 p mRNA in ARDS group was significantly lower than that in control group(P<0.01). The level of miR-34 b-5 p mRNA in ARDS+ miR-34 b-5 p mimics group was higher than that in ARDS group(P<0.01). The level of miR-34 b-5 p mRNA in lung tissue of ARDS+ miR-34 b-5 p mimics mock group was higher than that of ARDS group,but no significant difference(P>0.05). In the control group,the lung tissue was regular,complete and clear. In ARDS group,the structure of lung tissue was disordered,and a large number of red blood cells and inflammatory cells were found in alveoli.Compared with ARDS group,there was no significant difference in pathological changes of lung tissue in ARDS+ miR-34 b-5 p mimics mock group,but the lung tissue in ARDS+ miR-34 b-5 p mimics
作者
顾晓丽
陈芳
GU Xiao-li;CHEN Fang(Department of Pediatrics,the Second Affiliated Hospital of Xinjiang Medical University,Urumqi,Xinjiang 830063,China)
出处
《热带医学杂志》
CAS
2021年第6期705-710,F0003,共7页
Journal of Tropical Medicine
基金
新疆医科大学科研创新基金项目(XYDCX201632)。
