纳米氧化钕致SD大鼠肺部炎症反应中miR-498-5p的表达及意义  

The expression and significance of miR-498-5p in lung inflammation response of SD rats induced by nano-neodymium oxide

作　　者：陈世杰 赵鹏雅 孙丽颖 白钢[1] 黄丽华[1] CHEN Shi-jie;ZHAO Peng-ya;SUN Li-ying;BAI Gang;HUANG Li-hua(School of Public Healthy Baotou Medical College,Baotou,Inner Mongolia 014040,China)

机构地区：[1]包头医学院公共卫生学院,内蒙古包头014040

出　　处：《现代预防医学》2022年第7期1284-1288,1293,共6页Modern Preventive Medicine

基　　金：国家自然科学基金(81960596,81760600);内蒙古自治区自然科学基金(2019MS08057);内蒙古高等学校科学研究项目(NJZZ20170)。

摘　　要：目的 探讨纳米氧化钕(NPs-Nd_(2)O_(3))致SD大鼠肺部炎症反应中miR-498-5p的表达水平及意义。方法 将42只SPF级健康雄性SD大鼠随机分为对照组和NPs-Nd_(2)O_(3)染毒组,按照100 mg/kg浓度,采用非暴露式气管内一次性滴注法对大鼠进行NPs-Nd_(2)O_(3)染毒处理,在染毒第7、14和28 d后处死大鼠。计算肝脏、脑、肾脏、脾脏、睾丸、肺脏的脏器系数;采用电感耦合等离子体质谱法(ICP-MS)检测肝脏、脑、肾脏、脾脏、睾丸、肺脏中钕元素的含量;酶联免疫吸附法(ELISA)测定血清、肺泡灌洗液及肺组织中肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的含量;实时荧光定量PCR检测大鼠肺组织中miR-498-5p的相对表达水平;TargetScan、 mireap、miRanda网站预测与miR-498-5p相互作用的下游靶基因;Pearson相关分析miR-498-5p与炎症因子TNF-α、IL-6的相关性。结果 NPs-Nd_(2)O_(3)染毒后,大鼠肺脏的脏器系数增加(t=-3.408,P=0.005);肝脏、脑、肾脏、脾脏、睾丸、肺脏中均检测到钕元素;血清、肺泡灌洗液及肺组织中TNF-α、IL-6的含量升高(P<0.05);NPs-Nd_(2)O_(3)染毒第7、14和28 d后肺组织中miR-498-5p的相对表达水平升高(7 d组:t=-21.236,P<0.001;14 d组:t=-4.631,P=0.010;28 d组:t=-6.132,P=0.001),miR-498-5p表达与TNF-α含量呈正相关(r=0.787,P=0.012),与IL-6含量呈正相关(r=0.708,P<0.033);KEGG富集分析发现与miR-498-5p作用的靶基因其中有27个mRNAs参与JAK/STAT通路,19个mRNAs参与NF-κB通路。结论 NPs-Nd_(2)O_(3)经呼吸道进入体内可引起肺部明显炎症反应;NPs-Nd_(2)O_(3)染毒组大鼠肺组织中miR-498-5p表达水平升高,并与TNF-α、IL-6含量呈正相关;MiR-498-5p可能通过JAK/STAT通路和NF-κB通路参与NPs-Nd_(2)O_(3)诱导的肺部炎症。Objective To investigate the expression level and significance of miR-498-5 p in the lung inflammation of SD rats induced by nano neodymium oxide(NPs-Nd_(2)O_(3)). Methods A total of 42 SPF-grade healthy male SD rats were randomly divided into a control group and a NPs-Nd_(2)O_(3)exposure group. The rats were exposed to NPs-Nd_(2)O_(3)at a concentration of 100 mg/kg by a non-exposed intratracheal instillation method. The rats were sacrificed on the 7, 14 and 28 days after exposure. The organ coefficients of livers, brains, kidneys, spleens, testes and lungs were calculated. Icoupled plasma mass spectrometry(ICP-MS) was used to detect the content of neodymium in brains, kidneys, spleens, testes and lungs. Enzyme-linked immunosorbent assay(ELISA) was used to determine the levels of tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) in serums, alveolar lavage fluid and lung tissues. Real-time fluorescence quantitative PCR was used to detect the relative expression level of miR-498-5 p in rat lung tissues. TargetScan, mireap, miRanda software were used to predict downstream target genes that interact with miR-498-5 p. Pearson correlation was used to analyze the correlation between miR-498-5 p and inflammatory factors TNF-α and IL-6. Results After NPs-Nd_(2)O_(3)was exposed to poison, the organ coefficient of rat lungs increased(t=-3.408, P=0.005). Neodymium was detected in livers, brains, kidney, spleens, testis and lungs. IL-6 and TNF-α levels in serum, alveolar lavage fluid and lung tissue were increased(P<0.05). The relative expression level of miR-498-5 p in lung tissues increased on the 7, 14 and 28 days after NPs-Nd_(2)O_(3)exposure(7 d: t=-21.236, P<0.001;14 d: t=-4.631,P=0.010;28 d: t=-6.132,P=0.001), and the expression of miR-498-5 p was positively correlated with the levels of TNF-α(r=0.787, P=0.012) and IL-6(r=0.708, P<0.033). KEGG enrichment analysis found that among the target genes that interact with miR-498-5 p, 27 mRNAs were involved in the JAK/STAT pathway, and 19 mRNAs were involved in the

关 键 词：纳米氧化钕 IL-6 TNF-Α miR-498-5p 

分 类 号：R114[医药卫生—卫生毒理学]