异丙酚诱发新生大鼠海马神经元凋亡及与RhoA/ROCK2信号通路的关系
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摘要
目的探讨异丙酚诱发的新生大鼠海马神经元凋亡情况及其与RhoA/ROCK2信号通路的关系。方法 46只SD大鼠随机分为对照组,低、高剂量实验组,分别腹腔注射脂肪乳剂7. 5 mL·kg~(-1),异丙酚50 mg·kg~(-1),异丙酚100mg·kg~(-1)。分离并培养新生SD大鼠神经元,对照组(C组)神经元正常培养,低、高剂量异丙酚干预组(P组)神经元分别加入6,12μg·mL~(-1)异丙酚。用TUNEL法检测原代SD大鼠神经元凋亡情况,蛋白免疫印迹法检测大鼠海马组织中半胱氨酸蛋白酶-3(Caspase-3)及RhoA/ROCK2信号通路蛋白的表达。结果 C组,低、高剂量P组凋亡率分别为(9. 54±1. 13)%,(10. 53±1. 85)%,(27. 63±2. 54)%,高剂量异丙酚干预组分别与对照组和低剂量试验组比较,差异均有统计学意义(均P <0. 05)。对照组,低剂量实验组和高剂量实验组海马组织中Caspase-3蛋白相对表达量分别为0. 28±0. 03,0. 32±0. 05,0. 98±0. 25; RhoA蛋白相对表达量分别为0. 25±0. 05,0. 32±0. 06,0. 87±0. 08;ROCK2蛋白相对表达量分别为0. 59±0. 11,0. 63±0. 20,1. 53±0. 09,高剂量实验组分别与对照组和低剂量试验组比较,差异均有统计学意义(均P <0. 05)。结论高剂量异丙酚可诱发新生大鼠海马神经元凋亡,且RhoA/ROCK2信号通路的激活参与了该过程。
Objective To investigate the hippocampal neurons apoptosis induced by propofol in neonatal rats and its relationship with RhoA/ROCK2 signaling pathway. Methods Forty-six SD rats were randomly divided into control group and exp-L/-H group,which were injected intraperitoneally with 7. 5 mL·kg~(-1) of fat emulsion and 50,100 mg·kg~(-1) of propofol,respectively. Neurons of neonatal SD rats were isolated and cultured,neurons of control group( C group) were cultured normally,while neurons of propofol intervention( P-L/-H) group were given 6,12 μg·mL~(-1) of propofol,respectively. The apoptosis of primary neurons of SD rats was detected by TUNEL,and the expression of caspase-3 and RhoA/ROCK2 signaling pathway protein were detected by Western blot. Results In C group and P-L/-H group,apoptosis rates were( 9. 54 ± 1. 13) %,( 10. 53 ± 1. 85) %,( 27. 63 ± 2. 54) %,respectively,and the neuron apoptosis rate of the P-H group was significantly higher than those of the C group and P-L group( P < 0. 05).The expression of Caspase-3 protein in hippocampus of control group,exp-L/-H group were 0. 28 ± 0. 03,0. 32 ± 0. 05,0. 98 ± 0. 25,the expression of ROCK2 protein were 0. 25 ± 0. 05,0. 32 ± 0. 06,0. 87 ± 0. 08,the expression of ROCK2 protein were 0. 59 ± 0. 11,0. 63 ± 0. 20,1. 53 ± 0. 09,respectively,and the indicators of the exp-H group was significantly higher than those of the control group and exp-L group( P < 0. 05). Conclusion High-dose Propofol can induce apoptosis of hippocampal neurons in neonatal rats,and the activation of RhoA/ROCK2 signaling pathway is involved in this process.
Objective To investigate the hippocampal neurons apoptosis induced by propofol in neonatal rats and its relationship with RhoA/ROCK2 signaling pathway. Methods Forty-six SD rats were randomly divided into control group and exp-L/-H group,which were injected intraperitoneally with 7. 5 mL·kg~(-1) of fat emulsion and 50,100 mg·kg~(-1) of propofol,respectively. Neurons of neonatal SD rats were isolated and cultured,neurons of control group( C group) were cultured normally,while neurons of propofol intervention( P-L/-H) group were given 6,12 μg·mL~(-1) of propofol,respectively. The apoptosis of primary neurons of SD rats was detected by TUNEL,and the expression of caspase-3 and RhoA/ROCK2 signaling pathway protein were detected by Western blot. Results In C group and P-L/-H group,apoptosis rates were( 9. 54 ± 1. 13) %,( 10. 53 ± 1. 85) %,( 27. 63 ± 2. 54) %,respectively,and the neuron apoptosis rate of the P-H group was significantly higher than those of the C group and P-L group( P < 0. 05).The expression of Caspase-3 protein in hippocampus of control group,exp-L/-H group were 0. 28 ± 0. 03,0. 32 ± 0. 05,0. 98 ± 0. 25,the expression of ROCK2 protein were 0. 25 ± 0. 05,0. 32 ± 0. 06,0. 87 ± 0. 08,the expression of ROCK2 protein were 0. 59 ± 0. 11,0. 63 ± 0. 20,1. 53 ± 0. 09,respectively,and the indicators of the exp-H group was significantly higher than those of the control group and exp-L group( P < 0. 05). Conclusion High-dose Propofol can induce apoptosis of hippocampal neurons in neonatal rats,and the activation of RhoA/ROCK2 signaling pathway is involved in this process.
引文
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[5]闫少珍,王晓莉,王海宇,等.脐血单个核细胞侧脑室移植对HIBD新生大鼠脑神经元凋亡及Bax,Bcl-2蛋白的影响[J].中国当代儿科杂志,2016,18(9):862-866.
[6]CAPATI M L F,NAKAZONO A,IGAWA K,et al.Boron accelerates cultured osteoblastic cell activity through calcium flux[J].Biol Trace Elem Res,2016,174(2):300-308.
[7]楼烨亮,陈梦静,王可,等.哈巴苷对急性脑缺血及线粒体介导的Caspase依赖性细胞凋亡信号通路的影响[J].中国药理学通报,2017,33(4):563-567.
[8]高庆强,陈海,陈赟,等.Rock蛋白抑制剂法舒地尔对前列腺癌PC3,DU145细胞侵袭,迁移和凋亡的影响[J].中华男科学杂志,2016,22(6):483-490.
[2]WANG X,DING G,LAI W,et al.MicroRNA-383 upregulation protects against propofol-induced hippocampal neuron apoptosis and cognitive impairment[J].Exp Therap Med,2018,15(4):3181-3188.
[3]石咏梅,廖君,黄娟,等.内质网应激通路在脑泰方提取物保护局灶性缺血大鼠海马神经元中的作用[J].世界中医药,2016,11(4):587-591.
[4]明少鹏,周凤坤.抗氧化剂Mito Q对异氟醚诱导大鼠海马神经元损伤的影响及机制[J].临床麻醉学杂志,2017,33(10):1001-1005.
[5]闫少珍,王晓莉,王海宇,等.脐血单个核细胞侧脑室移植对HIBD新生大鼠脑神经元凋亡及Bax,Bcl-2蛋白的影响[J].中国当代儿科杂志,2016,18(9):862-866.
[6]CAPATI M L F,NAKAZONO A,IGAWA K,et al.Boron accelerates cultured osteoblastic cell activity through calcium flux[J].Biol Trace Elem Res,2016,174(2):300-308.
[7]楼烨亮,陈梦静,王可,等.哈巴苷对急性脑缺血及线粒体介导的Caspase依赖性细胞凋亡信号通路的影响[J].中国药理学通报,2017,33(4):563-567.
[8]高庆强,陈海,陈赟,等.Rock蛋白抑制剂法舒地尔对前列腺癌PC3,DU145细胞侵袭,迁移和凋亡的影响[J].中华男科学杂志,2016,22(6):483-490.