法舒地尔对蛛网膜下腔脑出血大鼠海马CA1区自噬作用及对mTOR通路的影响  

Effect of Fasudil on Autophagy in Hippocampal CA1 Region and mTOR Pathway in Rats with Subarachnoid Hemorrhage

作　　者：曹宏斌 冯乐霄 CAO Hong-bin;Feng Le-xiao(Shenmu City Hospital,Yulin City,Shaanxi Province,Shaanxi,Yulin,719300)

机构地区：[1]陕西省神木市医院,陕西榆林719300 [2]咸阳市中心医院,陕西咸阳712000

出　　处：《医学临床研究》2022年第1期18-22,共5页Journal of Clinical Research

摘　　要：【目的】探讨法舒地尔对蛛网膜下腔出血(SAH)大鼠海马CA1区自噬作用及对mTOR通路的影响。【方法】18只SPF级SD雄性大鼠随机分为假手术组(SHAM组,n=6)、SAH组(n=6)、法舒地尔组(FAS组,n=6)。FAS组于术后给予盐酸法舒地尔10mg/(kg·d)腹腔注射、连续3d;SHAM组、SAH组于同时间腹腔注射等量生理盐水,且SHAM组大鼠不刺破血管。采用穿梭箱实验检测各组大鼠学习记忆能力;采用苏木精-伊红(HE)染色观察各组大鼠海马CA1区神经细胞病理学变化,并计算正常神经细胞个数;采用免疫印迹法检测各组大鼠海马CA1区神经细胞自噬蛋白[微管相关蛋白轻链3(LC3)、家蚕隔离体蛋白1(p62)]及磷脂肌醇3-激酶/哺乳动物雷帕霉素靶蛋白(PI3K/mTOR)通路蛋白[PI3K、p-PI3K、mTOR、p-mTOR]蛋白变化。【结果】SHAM组、SAH组、FAS组大鼠逃避反应次数分别为(26.33±3.05)次、(18.52±2.03)次、(22.06±2.58)次,逃避反应时间分别为(3.69±1.01)s、(8.77±3.96)s、(6.05±2.51)s,正常神经细胞数分别为(125.69±18.85)个、(63.03±9.46)个、(95.77±14.33)个,LC3-Ⅱ/LC3-Ⅰ比值分别为(0.97±0.15)%、(0.38±0.06)%、(0.56±0.09)%,p62蛋白表达分别为0.45±0.07、1.13±0.17、0.81±0.13,p-PI3K/PI3K蛋白分别为0.65±0.09、1.58±0.24、1.03±0.15,p-mTOR/mTOR分别为0.73±0.10、1.36±0.20、1.11±0.17。与SHAM组比较,SAH组、FAS组大鼠逃避反应次数及CA1区组织正常神经细胞数、LC3-Ⅱ/LC3-Ⅰ比值减少或降低(P<0.05),逃避反应时间及CA1区组织p62蛋白、p-mTOR/mTOR蛋白、p-PI3K/PI3K蛋白水平增多或升高(P<0.05),但FAS组大鼠逃避反应次数及CA1区组织正常神经细胞数、LC3-Ⅱ/LC3-Ⅰ比值水平比SAH组增多或升高(P<0.05),逃避反应时间及CA1区组织p62蛋白、p-mTOR/mTOR蛋白、p-PI3K/PI3K蛋白水平比SAH组减少或降低(P<0.05)。【结论】法舒地尔可增强SAH大鼠海马CA1区神经细胞自噬对细胞发挥保护作用,进而增强大鼠学习记忆功能,其机�【Objective】To investigate the effect of fasudil on autophagy in hippocampal CA1 region and mTOR pathway in rats with subarachnoid hemorrhage(SAH).【Methods】Eighteen SPF male SD rats were randomly divided into sham operation group(SHAM group,n=6),SAH group(n=6)and fasudil group(FAS group,n=6).In FAS group,fasudil hydrochloride 10 mg/(kg?d)was injected intraperitoneally for 3 days;the SHAM group and the SAH group were intraperitoneally injected with the same amount of normal saline at the same time,and the rats in the sham group did not puncture the blood vessels.Shuttle box test was used to detect the learning and memory ability of rats in each group;Hematoxylin eosin(HE)staining was used to observe the pathological changes of neurons in hippocampal CA1 area of rats in each group,and the number of normal neurons was calculated;The changes of autophagy protein[tube associated protein light chain 3(LC3),silkworm isolate protein 1(p62)]and phosphoinositide 3-kinase/mammalian rapamycin target protein(PI3K/mTOR)pathway proteins[PI3K,p-PI3K,mTOR,p-mTOR]in hippocampal CA1 region of rats in each group were detected by Western blotting.【Results】The escape times of the sham group,the SAH group and the FAS group were(26.33±3.05),(18.52±2.03),(22.06±2.58)times,the escape time was(3.69±1.01)s,(8.77±3.96)s and(6.05±2.51)s,and the number of normal nerve cells was(125.69±18.85),(63.03±9.46)and(95.77±14.33)respectively,The ratio of LC3-Ⅱ/LC3-Ⅰwas(0.97±0.15)%,(0.38±0.06)%,(0.56±0.09)%,the expression of p62 protein was(0.45±0.07),(1.13±0.17),(0.81±0.13),p-PI3K/PI3K protein was 0.65±0.09,1.58±0.24,1.03±0.15,p-mTOR/mTOR was 0.73±0.10,1.36±0.20,1.11±0.17,respectively.Compared with the sham group,the number of escape reaction,the number of normal nerve cells in CA1 area,the ratio of LC3-Ⅱ/LC3-Ⅰand the level of p-PI3K/PI3K protein in the SAH group and the Fas group decreased or reduced(P<0.05),the escape reaction time and the levels of p62 protein and p-mTOR/mTOR protein in CA1 area increased or pr

关 键 词：蛛网膜下腔出血 血管舒张药/治疗应用 大鼠 疾病模型 动物 CA1区 海马/病理学 

分 类 号：R743.34[医药卫生—神经病学与精神病学]