摘要
目的探讨微RNA(miRNA)-210通过调控低氧诱导因子-1α(HIF-1α)/BNIP3/NIX通路和NOD样受体蛋白3(NLRP3)炎症小体对脂多糖诱导的新生大鼠缺氧缺血性脑损伤(HIBD)的保护作用及其作用机制。方法建立新生SD大鼠HIBD模型。将3日龄120只SD大鼠按照随机数字表法分为正常组、模型组、空载体组、miRNA-210过表达组、miRNA-210干扰组,每组各24只。正常组不做任何处理,正常饲养;模型组给予腹腔注射脂多糖,结扎左侧颈总动脉;空载体组给予腹腔注射空载体,miRNA-210过表达组给予腹腔注射miRNA-210过表达载体,miRNA-210干扰组给予腹腔注射miRNA-210干扰载体,然后按模型组方法处理。各组分别于造模后6、24、72 h各处死8只大鼠。检测并比较各组脑组织中miRNA-210、HIF-1α、Bcl-2/腺病毒E1B 19kDa相互作用蛋白3(BNIP3)、NIP3样蛋白X(NIX)、血管内皮生长因子(VEGF)、胰岛素样生长因子(IGF)-1 miRNA和蛋白的表达;检测并比较各组白细胞介素(IL)-1β、Caspase-1和NLRP3的表达水平;TUNEL染色评估各组神经元凋亡。结果造模后6、24、72 h,与正常组相比,模型组脑组织miRNA-210表达水平均降低,差异有统计学意义(P<0.05);与模型组相比,miRNA-210过表达组脑组织miRNA-210表达水平升高,差异均有统计学意义(P<0.05)。造模后24 h时,与正常组相比,模型组脑组织HIF-1α、BNIP3、NIX的mRNA和蛋白表达水平均升高,脑组织VEGF、IGF-1的mRNA和蛋白表达水平均降低,差异均有统计学意义(P<0.05);与模型组相比,miRNA-210过表达组HIF-1α、BNIP3、NIX的mRNA和蛋白表达水平均降低,脑组织VEGF、IGF-1的mRNA和蛋白表达水平升高,差异均有统计学意义(P<0.05)。造模后24 h时,与正常组相比,模型组脑组织IL-1β、Caspase-1、NLRP3水平均升高,差异均有统计学意义(P<0.05);与模型组相比,miRNA-210过表达组脑组织IL-1β、Caspase-1、NLRP3水平降低,差异均有统计学意义(P<0.05)。造模后6、24、72 h时,与正常组相比,模型组TUNEL阳性细胞数量均增多,差异有统计学意义(P<0.05);与模型组相比,miRNA-210过表达组TUNEL阳性细胞数量减少,差异均有统计学意义(P<0.05)。miRNA-210干扰组则表现相反。结论miRNA-210通过抑制HIF-1α/BNIP3/NIX通路和NLRP3炎症小体的活化,减少神经元凋亡,促进VEGF和IGF-1表达,从而减轻脂多糖诱导的新生大鼠HIBD。
Objective To investigate the protective effect and its mechanism of microRNA(miRNA)-210 on lipopolysaccharide-induced hypoxic-ischemic brain damage(HIBD)in neonatal rats through regulating hypoxia-inducible factor-1α(HIF-1α)/BNIP3/NIX pathway and NOD-like receptor protein 3(NLRP3)inflammasome.Methods The HIBD model of neonatal SD rats was established.According to the random number table method,1203-day-old SD rats were divided into the normal group,the model group,the empty vector group,the miRNA-210 overexpression group and the miRNA-210 interference group,with 24 rats in each group.The normal group did not do any treatment,normal feeding;the model group was given intraperitoneal injection of lipopolysaccharide,and the left common carotid artery was ligated.The empty vector group was given intraperitoneal injection of empty vector,the miRNA-210 overexpression group was given intraperitoneal injection of miRNA-210 overexpression vector,and the miRNA-210 interference group was given intraperitoneal injection of miRNA-210 interference vector,and then treated according to the model group method.Eight rats in each group were sacrificed at 6,24 and 72 h after mocleling respectively.The expression levels of miRNA-210,HIF-1α,Bcl-2/adenoriras E1B 19kDa interacting protein 3(BNIP3),vascular endothelial growth factor(VEGF)and insulin-like growth factor(IGF)-1 miRNA and protein,and NIP3-like protein X(NIX)in brain tissue were measured;the levels of interleukm(IL)-1β,Caspase-1,and NLRP3 were assessed;TUNEL staining was used to evaluate neuronal apoptosis.Results At 6,24 and 72 h after modeling,compared with the normal group,the expression level of miRNA-210 in the brain tissue of the model group were decreased,and the differences were statistically significant(P<0.05).Compared with the model group,the expression level of miRNA-210 in the brain tissue of the miRNA-210 overexpression group wwere increased,and the differences were statistically significant(P<0.05).At 24 h after modeling,compared with the normal group,the mRNA and protein expression levels of HIF-1α,BNIP3 and NIX in the brain tissue of the model group were increased,and the mRNA and protein expression levels of VEGF and IGF-1 in the brain tissue of the model group were decreased,the differences were statistically significant(P<0.05).Compared with the model group,the mRNA and protein expression levels of HIF-1α,BNIP3 and NIX in the brain tissue of the miRNA-210 overexpression group were decreased,and the mRNA and protein expression levels of VEGF and IGF-1 in the brain tissue of the miRNA-210 overexpression group were increased,the differences were statistically significant(P<0.05).At 24 h after modeling,compared with the normal group,the levels of IL-1β,Caspase-1 and NLRP3 in the brain tissue of the model group were increased,and the differences were statistically significant(P<0.05).Compared with the model group,the levels of IL-1β,Caspase-1 and NLRP3 in the brain tissue of the miRNA-210 overexpression group were decreased,and the differences were statistically significant(P<0.05).At 6,24 and 72 h after modeling,compared with the normal group,the number of TUNEL positive cells in the model group increased,and the difference was statistically significant(P<0.05).Compared with the model group,the number of TUNEL positive cells in the miRNA-210 overexpression group decreased,and the difference was statistically significant(P<0.05).Conversely,in the miRNA-210 interference group,these parameters showed opposite trends.Conclusion miRNA-210 alleviates lipopolysaccharide-induced HIBD in neonatal rats by inhibiting the activation of HIF-1α/BNIP3/NIX pathway and NLRP3 inflammasome,reducing neuronal apoptosis,and promoting the expression of VEGF and IGF-1.
作者
聂晶
张薇薇
杨舒贺
梅梅
NIE Jing;ZHANG Weiwei;YANG Shuhe(Department of Pediatrics,The First Affiliated Hospital of Jiamusi University,Jiamusi Heilongjiang 154002,China;Jiamusi University,Jiamusi Heilongjiang 154002,China)
出处
《临床和实验医学杂志》
2025年第22期2357-2361,共5页
Journal of Clinical and Experimental Medicine
基金
黑龙江省教育厅基本科研业务费基础研究项目(编号:2021-kyywf-0602)
佳木斯大学附属第一医院国家重点专科建设项目团队:儿科科研团队项目(编号:GJ202301)。
