miR-4472靶向PIN1调控NF-κB/STAT3信号通路在缺氧缺血性脑病大鼠模型中的作用及机制研究

Role and mechanism of miR-4472 targeting PIN1 in regulating the NF-κB/STAT3 signaling pathway in a rat model of hypoxic-ischemic encephalopathy

目的探讨微小RNA(miR)-4472靶向脯氨酸异构酶(PIN1)调控核转录因子(NF)-κB/信号传导子和转录激活子3(STAT3)信号通路在缺氧缺血性脑病大鼠模型(HIE)中的作用及机制。方法究主导单位为嘉兴市第二医院,研究时间为2022年1月至2024年1月。选取Spargue Dawley(SD)大鼠60只,依据随机数字表法将其随机分为六组,各组间体质量差异均无统计学意义(均P>0.05)。分别为正常对照组、HIE模型组、抑制对照组、miR-4472抑制组、抑制miR-4472+干扰对照组、抑制miR-4472+干扰PIN1组,每组10只。采用Rice-Vannucci法建立HIE模型。采用Morris水迷宫实验评价大鼠行为学;采用Longa评分法评价神经功能;采用TUNEL检测凋亡;采用Western blot测定NF-κB和STAT3蛋白表达。结果与HIE模型组比较,miR-4472抑制组和抑制miR-4472+干扰对照组大鼠逃避潜伏期缩短,而抑制miR-4472+干扰PIN1组大鼠逃避潜伏期延长(P<0.05)。大鼠穿越平台次数比较,抑制miR-4472+干扰PIN1组[(2.13±0.54)次]低于模型组[(3.56±0.71)次]、抑制对照组[(3.61±0.87)次]、miR-4472抑制组[(5.47±1.29)次]和抑制miR-4472+干扰对照组[(5.58±1.32)次](t=5.07、4.57、7.55、7.65,均P<0.05)。大鼠Longa评分比较,抑制miR-4472+干扰PIN1组[(3.03±0.30)分]低于HIE模型组[(2.45±0.54)分]、抑制对照组[(2.38±0.69)分]、miR-4472抑制组[(1.27±0.46)分]和抑制miR-4472+干扰对照组[(1.29±0.51)分](t=2.97、2.73、10.13、9.30,均P<0.05)。大鼠海马神经凋亡率比较,抑制miR-4472+干扰PIN1组[(25.34±6.16)%]低于HIE模型组[(18.42±5.46)%]、抑制对照组[(17.95±4.38)%]、miR-4472抑制组[(8.89±2.10)%]和抑制miR-4472+干扰对照组[(9.13±2.57)%](t=2.97、2.73、10.13、9.30,均P<0.05)。与HIE模型组比较,miR-4472抑制组和抑制miR-4472+干扰对照组大鼠NF-κB和STAT3蛋白灰度值降低,而抑制miR-4472+干扰PIN1组大鼠NF-κB和STAT3蛋白灰度值增加(均P<0.05)。结论miR-4472靶向调控PIN1促进HIE损伤,其机制与激活NF-κB/STAT3信号通路有关。

ObjectiveTo investigate the role and mechanism of microRNA(miR)-4472 targeting PIN1 in regulating the nuclear factor kappa B(NF-κB)/signal transducer and activator of transcription 3(STAT3)signaling pathway in a rat model of hypoxic-ischemic encephalopathy(HIE).MethodsBetween January 2022 and January 2024,sixty Sprague-Dawley rats were randomly assigned to six groups at The Second Hospital of Jiaxing using a random number table method:a normal control group,an HIE model group,an inhibition control group,a miR-4472 inhibition group,a miR-4472 inhibition+interference control group,and a miR-4472 inhibition+PIN1 interference group,with ten rats in each group.There was no significant difference in body mass among the six groups(all P>0.05).Rat models of HIE were established using the Rice-Vannucci method.Behavioral performance was assessed using the Morris water maze test,while neurological function was evaluated using the Longa scoring method.Apoptosis was detected using the TUNEL assay,and the expression of NF-κB and STAT3 protein was measured using Western blot analysis.ResultsCompared with the HIE model group,the miR-4472 inhibition group and the miR-4472 inhibition+interference control group showed a shortened escape latency,while the miR-4472 inhibition+PIN1 interference group exhibited an extended escape latency(all P<0.05).The number of platform crossings in the miR-4472 inhibition+PIN1 interference group[(2.13±0.54)times]was significantly lower than that in the HIE model group[(3.56±0.71)times],the inhibition control group[(3.61±0.87)times],the miR-4472 inhibition group[(5.47±1.29)times],and the miR-4472 inhibition+interference control group[(5.58±1.32)times](t=5.07,4.57,7.55,7.65,all P<0.05).The Longa score in the miR-4472 inhibition+PIN1 interference group[(3.03±0.30)points]was significantly lower than that in the HIE model group[(2.45±0.54)points],the inhibition control group[(2.38±0.69)points],the miR-4472 inhibition group[(1.27±0.46)points],and the miR-4472 inhibition+interference control group[(1.29±0.51)points](t=2.97,2.73,10.13,9.30,all P<0.05).The apoptosis rate of hippocampal neurons in the miR-4472 inhibition+PIN1 interference group[(25.34±6.16)%]was significantly lower than that in the HIE model group[(18.42±5.46)%],the inhibition control group[(17.95±4.38)%],the miR-4472 inhibition group[(8.89±2.10)%],and the miR-4472 inhibition+interference control group[(9.13±2.57)%](t=2.97,2.73,10.13,9.30,all P<0.05).Compared with the HIE model group,the miR-4472 inhibition group and the miR-4472 inhibition+interference control group exhibited decreased gray values of NF-κB and STAT3 protein,while the miR-4472 inhibition+PIN1 interference group showed increased gray values of NF-κB and STAT3 protein(all P<0.05).ConclusionmiR-4472 targets and regulates PIN1,which contributes to HIE injury through the activation of the NF-κB/STAT3 signaling pathway.