摘要
目的探讨微小RNA(miRNA,miR)-96如何通过胰岛素样生长因子1类受体(IGF1R)调控七氟醚诱导新生大鼠认知功能障碍。方法将大鼠(购自上海斯莱克实验动物有限公司)随机分为6组,对照组大鼠吸入浓度为40%的O2,实验组小鼠吸入不同浓度的七氟烷麻醉(购自上海雅培上海有限公司),包括1%七氟醚组,2%七氟醚组,4%七氟醚组,4%七氟醚+miR-96过表达组和4%七氟醚+miR-96抑制组。通过给予不同浓度的七氟醚(1%、2%和4%)联合miR-96激动或抑制剂,在mRNA和蛋白水平检测IGF1R、B细胞淋巴瘤/白血病-2(bcl-2)、bcl-2相关X蛋白(bax)、半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3的表达,测定海马神经元凋亡情况。通过测定荧光素酶活性,验证miR-96是否靶向作用于IGF1R。采用Morris水迷宫测验评价学习记忆能力。测量数据用均值±标准差(Mean±SD)表示,采用t检验对正态分布和均方差的数据进行分析,在双因素相关分析中,则采用Spearman秩相关。结果对照组、4%七氟醚组、4%七氟醚+miR-96过表达组和4%七氟醚+miR-96抑制组miR-96相对表达量分别为0.37±0.01、2.09±0.06、2.84±0.12、1.71±0.03,IGF1R相对表达量分别为2.64±0.08、0.51±0.06、0.29±0.02、1.25±0.03,吸入七氟醚后miR-96表达显著增加(t=63.230、45.870、94.750,P<0.05),IGF1R表达显著降低(t=47.630、63.720、36.380,P<0.05),差异有统计学意义。与4%七氟醚组比较,4%七氟醚+miR-96过表达组miR-96表达增强(t=12.500,P<0.05),IGF1R表达降低(t=7.780,P<0.05);而在4%七氟醚+miR-96抑制剂组中则相反(t=12.670、24.670,P<0.05),差异有统计学意义。结论七氟醚麻醉诱导新生大鼠发生认知功能障碍的潜在机制与miR-96通过抑制IGF1R调控有关。
Objective To investigate the relationship between sevoflurane and microRNA(miRNA,miR)-96,in an attempt to identify the means by which it mediates insulin like growth factor-1 receptor(IGF1R)to influence the cognitive dysfunction(CD)in neonatal rats.Relationship between differentially expressed miRNAs and sevoflurane concentration was identified.Methods Rats were randomly divided into 6 groups,including 1%sevoflurane group,2%sevoflurane group,4%sevoflurane group,4%sevoflurane+miR-96 mimic group and 4%sevoflurane+miR-96 inhibitor group.The animlas were given varying concentrations of the agent(1%,2%and 4%),combined with miR-96 mimic or an inhibitor.The mRNA and protein expression levels of IGF1R,B cell lymphoma/leukemia-2(bcl-2),bcl-2 associated X protein(bax),and cysteinyl aspartate-specific protease(Caspase)-3 were detected,and apoptosis of hippocampal neurons was measured.Learning and memory performance was assessed using the Morris water maze(MWM)test and step-down test.The measured data were expressed as mean±standard deviation.T-test was used to analyze the data of normal distribution and mean square deviation.In the two factor correlation analysis,Spearman rank correlation was used.P<0.05 was set as statistically significant difference.Results The relative expression of miR-96 in control group,4%sevoflurane group,4%sevoflurane+miR-96 mimic group and 4%sevoflurane+miR-96 inhibitor group was(0.37±0.01),(2.09±0.06),(2.84±0.12),(1.71±0.03),and the relative expression of IGF1R was(2.64±0.08),(0.51±0.06),(0.29±0.02),(1.25±0.03),respectively.After inhalation of sevoflurane,the expression of miR-96 was significantly increased(t=63.230,45.870,94.750,P<0.05),and the expression of IGF1R was significantly decreased(t=47.630,63.720,36.380,P<0.05).As compared with the 4%sevoflurane group,the expression of miR-96 in the 4%sevoflurane+miR-96 mimic group increased(t=12.500,P<0.05),and the expression of IGF1R decreased(t=7.780,P<0.05),while in the 4%sevoflurane+miR-96 inhibitor group,there was the opposite trend(t=12
作者
徐畅
牛姣姣
周俊飞
艾艳秋
Xu Chang;Niu Jiaojiao;Zhou Junfei;Ai Yanqiu(Department of Anesthesiology,Pain and Perioperative Medicine,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2020年第2期265-267,共3页
Chinese Journal of Experimental Surgery
