摘要
目的 探讨七氟烷吸入对β淀粉样蛋白(Aβ)_(1-40)诱发的大鼠认知功能障碍影响及其可能的机制。方法 于2021年8月至2022年8月,将32只成年雄性Sprague-Dawley大鼠以随机数字表法分为生理盐水(NS)+氧气组、NS+七氟烷组、Aβ+氧气组和Aβ+七氟烷组,每组8只,分别给予双侧海马内注射NS或Aβ_(1-40)。吸入30%氧气或2.5%七氟烷过程中监测生命体征,采用Morris水迷宫实验检测大鼠认知功能;酶联免疫吸附测定检测海马Aβ_(1-40)水平;免疫组织化学法检测胶质纤维酸性蛋白(GFAP)和离子钙接头蛋白分子1(IBA1)的表达;实时荧光定量逆转录聚合酶链反应检测白细胞介素-1β(IL-1β)、核因子-κB(NF-κB)、诱导型一氧化氮合酶(iNOS)的mRNA表达;蛋白质印迹法检测B淋巴细胞瘤-xL(Bcl-xL)、胱天蛋白酶-9(caspase-9)、脑源性神经营养因子(BDNF)和晚期糖基化终末产物受体(RAGE)的蛋白表达。结果 维持吸入2.5%七氟烷在4个不同时间点(1、2、3和4 h)对大鼠生命体征无影响。与NS+氧气组比较,Aβ+氧气组大鼠原始平台探索时间减少,逃逸潜伏期增加;与Aβ+氧气组相比,Aβ+七氟烷组大鼠原始平台探索时间减少,逃逸潜伏期增加(P<0.05)。Aβ+七氟烷组大鼠海马区Aβ_(1-40)水平[(42.18±5.72)ng/L],GFAP和IBA1阳性细胞数[(24.33±1.21)个和(37.82±3.23)个],caspase-9和RAGE蛋白表达(0.74±0.11和0.81±0.07),IL-1β、NF-κB和iNOS mRNA表达(28.98±12.32、25.91±12.31和43.92±17.21)均高于NS+七氟烷组[(18.13±2.89)ng/L、(10.51±0.96)个、(17.17±1.77)个、0.23±0.02、0.29±0.03、1.35±0.04、1.37±0.05、1.42±0.06]和Aβ+氧气组[(32.61±4.82)ng/L、(15.76±1.25)个、(24.76±2.31)个、0.45±0.08、0.68±0.08、10.23±6.56、6.51±2.34、13.23±4.81];Bcl-xL和BDNF蛋白表达(0.13±0.04和0.18±0.04)低于NS+七氟烷组(1.07±0.31和0.58±0.07)和Aβ+氧气组(0.46±0.11和0.33±0.04)(P<0.05)。而NS+氧气组和NS+七氟烷组之间各指标差异无统计学意义(P>0.05)。结论 七氟烷通过启动大鼠海马的神经毒性、神经炎症和神经元凋亡,加剧了Aβ_(1-40)诱发的大鼠认知功能障碍。
Objective To investigate the effects of sevoflurane inhalation on cognitive dysfunction induced by β Amyloid protein_(1-40)in rats and its possible mechanism.Methods From August 2021 to August 2022,32 adult male Sprague-Dawley rats were divided into normal saline(NS)+oxygen group,NS+sevoflurane group,Aβ+oxygen group,and Aβ+sevoflurane group by random number table method,with 8 rats in each group,and were given bilateral intra-hippocampal injection of NS or Aβ_(1-40),respectively.Vital signs were monitored during inhalation of 30% oxygen or 2.5% sevoflurane,and cognitive function was detected by Morris water maze test.The levels of Aβ_(1-40) in hippocampus were detected by enzyme-linked immunosorbent assay.The expressions of glial fibrillary acidic protein(GFAP) and IBA1 were detected by immunohistochemistry.The mRNA expression of interleukin-1β(IL-1β),nuclear factor-κB(NF-κB) and inducible nitric oxide synthase(iNOS) was detected by real-time fluorescence quantitative polymerase chain reaction.Protein expression of B lymphoblastoma xL(Bcl-xL),cysteine-containing aspartate protein hydrolase-9(caspase-9),brain-derived neurotrophic factor(BDNF),and advanced glycosylated end-product receptor(RAGE) were detected by Western blotting.Results Inhalation of 2.5% sevoflurane at 4 different time points(1,2,3 and 4h) had no effect on the vital signs of rats.Compared with NS+oxygen group,the exploration time of the original platform was reduced and the escape latency was increased in Aβ+oxygen group.Compared with Aβ+oxygen group,the original platform exploration time of rats in Aβ+sevoflurane group decreased and the escape latency increased(P<0.05).The levels of Aβ_(1-40) [(42.18±5.72) ng/L],the number of GFAP and IBA1 positive cells(24.33±1.21 and 37.82±3.23),the expression of caspase-9 and RAGE protein(0.74±0.11 and 0.81±0.07),the mRNA expressions of IL-1β,NF-κB and iNOS(28.98±12.32,25.91±12.31,and 43.92±17.21) in Aβ+sevoflurane group were higher than those in NS+sevoflurane group [(18.13±2.89) ng/L,10.51±0.96,17.17±1.77,0.23±0.02,0.29±0.03,1.35±0.04,1.37±0.05,1.42±0.06] and Aβ +oxygen group [(32.61±4.82) ng/L,15.76±1.25,24.76±2.31,0.45±0.08,0.68±0.08,10.23±6.56,6.51±2.34,13.23±4.81],the protein expressions of Bcl-xL and BDNF(0.13±0.04 and 0.18±0.04) were lower than those of NS+sevoflurane group(1.07±0.31 and 0.58±0.07) and Aβ +oxygen group(0.46±0.11 and 0.33±0.04)(P<0.05).There was no significant difference between NS+oxygen group and NS+sevoflurane group(P>0.05).Conclusion Sevoflurane can aggravate the cognitive dysfunction induced by Aβ_(1-40) by activating neurotoxicity,neuroinflammation and neuronal apoptosis in rat hippocampus.
作者
邹敏
孙应中
魏艳妮
官焕春
ZOU Min;SUN Yingzhong;WEI Yanni;GUAN Huanchun(Department of Anesthesiology,Chongqing Kaizhou District People's Hospital,Chongqing 405400,China)
出处
《安徽医药》
CAS
2024年第3期456-461,共6页
Anhui Medical and Pharmaceutical Journal
基金
重庆市2018年第七批科技计划项目(cstc2018jcyjAX0412)。
