七叶莲花醇提物通过调节PI3K/Akt/GSK-3β信号通路减轻丙泊酚所致新生大鼠神经毒性  

Ethanolic extract of Lotus esculentus alleviates neonatal rats neurotoxicity induced by regulating the PI3K/Akt/GSK-3βsignaling pathway

作　　者：杜引 马晓真 王龙[1] 袁浩峥 DU Yin;MA Xiao-zhen;WANG Long(Department of Anesthesiology,the Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an Shaanxi 710004,China;Department of Physiology,School of Basic Medicine,Shaanxi University of Traditional Chinese Medicine,Xianyang Shaanxi 712046,China)

机构地区：[1]西安交通大学第二附属医院麻醉科,陕西西安710004 [2]陕西中医药大学基础医学院生理教研室,陕西咸阳712046

出　　处：《临床和实验医学杂志》2022年第21期2249-2253,共5页Journal of Clinical and Experimental Medicine

基　　金：陕西省自然科学基础研究项目(编号:2021JQ-740);陕西省自然科学基础研究项目(编号:2022JQ-880)。

摘　　要：目的分析七叶莲花醇提物调节磷脂酰肌醇3-激酶/丝氨酸苏氨酸激酶/糖原合成酶激酶-3β(PI3K/Akt/GSK-3β)信号通路减轻丙泊酚所致新生大鼠神经毒性机制。方法清洁级健康雄性SD大鼠85只,采用随机数字表法分为5组,每组17只,对照组灌胃0.9%氯化钠溶液,阳性药组(给予丙泊酚),3种不同剂量七叶莲花醇提物组:低剂量组(3.06 g/kg)、中剂量组(6.12 g/kg)、高剂量组(12.24 g/kg),均灌胃七叶莲花醇提物,连续3 d,每天注射1次,腹腔注射戊巴比妥钠,通过肢体Ⅱ型导联监测心电图显露股动脉,置入24 G套管针抽血样,显露大鼠股静脉插入套管针泵注丙泊酚,速度为2 mg·kg^(-1)·min^(-1)。离断颈椎处死解剖分离大鼠海马组织,采用TUNEL染色检出海马神经元凋亡水平,RT-聚合链反应和蛋白质印迹法检测海马组织N-甲基-D-天冬氨酸1(NMDA1)、NMDA2B表达;RT-聚合链反应检测PI3K、Akt、GSK-3β的微小核糖核酸(mRNA)表达,蛋白质印迹法检测Akt、pAkt(ser473)、GSK-3β、PGSK-3β(ser9)蛋白表达。结果低、中、高剂量组及阳性药组海马神经元凋亡率均低于对照组,差异有统计学意义(P<0.05)。中剂量组、高剂量组及阳性药组NMDA1、NMDA2B mRNA表达与两者蛋白表达均低于对照组,差异均有统计学意义(P<0.05);低剂量组、中剂量组、高剂量组NMDA1、NMDA2B mRNA表达与两者蛋白表达高于阳性药组,差异均有统计学意义(P<0.05)。阳性药组GSK-3βmRNA表达高于对照组,高剂量组GSK-3βmRNA表达低于阳性药组,差异均有统计学意义(P<0.05)。低、中、高剂量组及阳性药组pAkt(ser473)、PGSK-3β(ser9)蛋白表达、pAkt(ser473)/Akt比值低于对照组,低、中剂量组及阳性药组PGSK-3β(ser9)/GSK-3β比值低于对照组,差异均有统计学意义(P<0.05);高剂量组pAkt(ser473)蛋白、pAkt(ser473)/Akt比值高于阳性药组,低、中、高剂量组PGSK-3β(ser9)蛋白、PGSK-3β(ser9)/GSK-3β比值高于阳性药组,差异均有�Objective To analyze the mechanism of ethanolic extract of Lotus esculentus regulating phosphatidylinositol 3-kinase/protein serine threonine kinase/glycogen synthase kinase 3-β(PI3K/Akt/GSK-3β)signaling pathway to reduce propofol-induced neurotoxicity in neonatal rats.Methods Eighty-five healthy male Sprague Dawley rats were randomly divided into 5 groups,with 17 rats in each group,a control group(intragastric normal saline),positive group(propofol administration),3 groups of different doses of Lotus esculentus ethanolic extract[the low dose group(3.06 g/kg),the medium dose group(6.12 g/kg),the high dose group(12.24 g/kg),all was given intragastric administration of Lotus esculentus ethanolic extract]and the positive drug group,once a day for 3 days.Anesthesia was induced by intraperitoneal injection of pentobarbital sodium.The femoral artery was exposed by electrocardiogram(ECG)monitoring in type II leads of the limbs.A 24 G trocar was placed to draw blood samples.The femoral vein of rats was exposed and trocar was inserted into the trocar pump to inject propofol at a rate of 2 mg·kg^(-1)·min^(-1).The hippocampal tissues were dissected and separated by cervical spine transection.The apoptosis level of hippocampal neurons was detected by TUNEL staining.RT-PCR and Western blotting were used to detect N-methyl-D-aspartate 1(NMDA1)and NMDA2B expression in hippocampal tissue;RT-PCR was used to detect microribonucleic acid(mRNA)expression of PI3K,Akt,and GSK-3β,and Western blot was used to detect mRNA expression of Akt,pAkt(ser473),GSK-3β,and PGSK-3β(ser9).Results The apoptosis of hippocampal neurons in the low dose group,the medium dose group and the high dose group was lower than that in the control group,the difference was statistically significant(P<0.05).The expression of NMDA1 and NMDA2B mRNA and their protein expressions in the medium dose group,the high dose group and the positive drug group were lower than those in the control group,the differences were statistically significant(P<0.05);the expression

关 键 词：新生儿 大鼠 七叶莲花醇提物 PI3K Akt GSK-3 丙泊酚 神经毒性 

分 类 号：R285.5[医药卫生—中药学]