天麻素对癫痫大鼠p- AKT和caspase3的影响
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摘要
目的观察天麻素对戊四氮诱导癫痫模型大鼠p-AKT和caspase 3的影响。方法将69只Wistar大鼠分为空白对照组18只、癫痫模型组28只(PTZ组,pentylenetetrazole,55～75 mg/kg)、天麻素干预组23只(Gas组,Gastrodin,200 mg/kg),除空白对照组外均腹腔注射戊四氮(PTZ)诱导形成癫痫模型,天麻素干预组给予天麻素(Gas)干预。电镜下观察各组大鼠海马组织形态学变化,免疫组化及Western blot检测各组大鼠海马组织丝氨酸/苏氨酸蛋白激酶(p-AKT)、含半胱氨酸的天冬氨酸蛋白水解酶(caspase 3)蛋白的表达。结果与对照组比较,PTZ组海马区表现为明显的癫痫病理改变,神经元变性及坏死显著,主要为大锥体细胞和部分齿状回颗粒细胞,损伤细胞细胞核固缩,核碎裂样改变,胞浆肿胀,尼氏体消失,细胞周隙增大,体积增大。药物干预后病理改变改善。大鼠癫痫发作后,癫痫模型组p-AKT、caspase 3蛋白表达高于空白对照组(P<0.05)。与癫痫模型组比较,天麻素干预组caspase 3的表达下调(P<0.05),在第3天差异尤为显著(P<0.01)。结论天麻素通过减少p-AKT、caspase 3蛋白表达,起到抗凋亡的作用,对癫痫模型起到一定保护作用。
Objective To observe the effect of gastrodine on p-AKT and caspase 3 in rats with epilepsy induced by pentylenetetrazole(PTZ). Methods69 Wistar rats were assigned into blank control group(18 rats), epilepsy model group(PTZ group, pentylenetetrazole, 55～75 mg/kg)(28 rats) and gastrodine group(Gas group, gastrodine, 200 mg/kg)(23 rats). Epileptic model was established with rats in all of groups by PTZ intraperitoneal injection except the blank control group. Gastrodine was given to the Gas group. Histomorphological changes of hippocampus were observed under electron microscope. The expression of serine-threonine protein kinase(p-AKT) and cysteine-aspartic acid specific protease 3(caspase 3) in hippocampus was detected by immunohistochemistry and Western blotting. Results Compared with the control group, the hippocampal area of the PTZ group showed remarkable pathological changes of epilepsy, including significant neuronal degeneration and necrosis, mainly the large pyramidal cells and some dentate gyrus granulosa cells, nucleus condensation in damaged cells, fragmentation-like nuclear changes, cytosolic swelling, disappearance of the nissl bodies and increase in cell peripheral gap and cell volume. The pathological changes were improved after drug intervention. After seizure in rats, the expression of p-AKT and caspase-3 was higher in the epilepsy model group than in the blank control group(P<0.05). Compared with the epilepsy model group, the expression of caspase-3 decreased in the gastrodine group(P<0.05), with very significant difference at day 3 between the two groups(P<0.01). Conclusion Gastrodine plays an anti-apoptotic role and protects against epilepsy by reducing the expression of p-AKT and caspase 3.
Objective To observe the effect of gastrodine on p-AKT and caspase 3 in rats with epilepsy induced by pentylenetetrazole(PTZ). Methods69 Wistar rats were assigned into blank control group(18 rats), epilepsy model group(PTZ group, pentylenetetrazole, 55～75 mg/kg)(28 rats) and gastrodine group(Gas group, gastrodine, 200 mg/kg)(23 rats). Epileptic model was established with rats in all of groups by PTZ intraperitoneal injection except the blank control group. Gastrodine was given to the Gas group. Histomorphological changes of hippocampus were observed under electron microscope. The expression of serine-threonine protein kinase(p-AKT) and cysteine-aspartic acid specific protease 3(caspase 3) in hippocampus was detected by immunohistochemistry and Western blotting. Results Compared with the control group, the hippocampal area of the PTZ group showed remarkable pathological changes of epilepsy, including significant neuronal degeneration and necrosis, mainly the large pyramidal cells and some dentate gyrus granulosa cells, nucleus condensation in damaged cells, fragmentation-like nuclear changes, cytosolic swelling, disappearance of the nissl bodies and increase in cell peripheral gap and cell volume. The pathological changes were improved after drug intervention. After seizure in rats, the expression of p-AKT and caspase-3 was higher in the epilepsy model group than in the blank control group(P<0.05). Compared with the epilepsy model group, the expression of caspase-3 decreased in the gastrodine group(P<0.05), with very significant difference at day 3 between the two groups(P<0.01). Conclusion Gastrodine plays an anti-apoptotic role and protects against epilepsy by reducing the expression of p-AKT and caspase 3.
引文
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[21] SHAO H,YANG Y,QI A P,et al.Gastrodin reduces the severity of status epilepticus in the rat pilocarpine model of temporal lobe epilepsy by inhibiting nav1.6 sodium currents[J].Neurochem Res,2017,42(2):360- 374
[22] 夏烈新,陈立铭,彭晓艳,等.天麻素对癫痫大鼠海马神经元IL1、TNF及IL10的影响[J].临床神经病学杂志,2017,30(3):209- 213.
[2] PITK?NEN A,ENGEL J J R.Past and present definitions of epileptogenesis and its biomarkers[J].Neurotherapeutics,2014,11(2):231- 241.
[3] STAFSTROM C E, CARMANT L.Seizures and epilepsy:an overview for neuroscientists[J].Cold Spring Harb Perspect Med,2015,5(6):a022426.
[4] LIU Y,GAO J,PENG M,et al.A review on central nervous system effects of gastrodin[J].Front Pharmacol,2018,9(24):1- 18.
[5] 孟丽,孟杰,刘云霞,等.戊四氮诱导癫痫大鼠发作对大鼠海马神经元的影响[J].现代预防医学,2007,34(16):3051- 3055.
[6] HUANG X H , ZHANG H L, YANG J, et al.Pharmacological inhibition of the mammalian target of rapamycin pathway suppresses acquired epilepsy[J].Neurobio Dise,2010,40(1):193- 199.
[7] RACINE R J.Mondification of seizure activity by electrical stimulation.II.Motor seizure[J].Electroencephalogr Clin Neurophysiol, 1972, 32(3):281- 294.
[8] HENSHALL D C, SIMON R P.Epilepsy and apoptosis pathways[J].J Cereb Blood Flow Metab,2005,25(12):1557- 1572.
[9] STRASSER A, O’CONNOR L, DIXIT V M.Apoptosis signaling[J].Annu Rev Biochem,2000(69):217- 245.
[10] HENSHALL D C.Apoptosis signalling pathways in seizure- induced neuronal death and epilepsy[J]Biochem Soc Trans,2007,35(Pt 2) :421- 423.
[11] YU J S, CUI W.Proliferation,survival and metabolism:the role of PI3K/AKT/mTOR signalling in pluripotency and cell fate determination[J].Development,2016,143(17):3050- 3060.
[12] SALE E M,SALE G J.Protein kinase B: signalling roles and therapeutic targeting[J].Cell Mol Life Sci,2008,65(1):113- 127.
[13] WANG L,ZHOU K,FU Z,et al.Brain development and Akt signaling: the crossroads of signaling pathway and neurodevelopmental diseases[J].J Mol Neurosci,2017,61(3):379- 384.
[14] 尹承龙,劳学军.PI3K- AKT- mTOR信号通路的研究进展[J].中国医学创新,2016,13(1):145- 148.
[15] L?SCHER W.Critical review of current animal models of seizures and epilepsy used in the discovery and development of new antiepileptic drugs[J].Seizure,2011,20(5):359- 368.
[16] CAMPOS G, FORTUNA A, FALC?O A, et al.In vitro and in vivo experimental models employed in the discovery and development of antiepileptic drugs for pharmacoresistant epilepsy[J]. Epilepsy Res, 2018(146):63- 86.
[17] JANDOVá K,RILJAK V,POKORN ,et al.Pentylentetrazol associated changes of hippocampal neurons in immature rats[J].Prague Med Rep,2017,108(1):67- 74.
[18] BIHAQI S W,ALANSI B,MASOUD A M,et al.Influence of early life lead (Pb) exposure on α- Synuclein,GSK- 3β and caspase- 3 mediated tauopathy:implications on Alzheimer’s disease[J].Curr Alzheimer Res,2018,15(12):1114- 1122.
[19] LIU B,LI F,SHI J,et al.Gastrodin ameliorates subacute phase cerebral ischemia- reperfusion injury by inhibiting inflammation and apoptosis in rats[J].Mol Med Rep,2016,14(5):4144- 4152.
[20] 秦瑞怡,边立功,钟莲梅,等.天麻素调控Nrf2信号通路在癫痫中的抗炎作用[J].神经解剖学杂志,2018,34(2):147- 152.
[21] SHAO H,YANG Y,QI A P,et al.Gastrodin reduces the severity of status epilepticus in the rat pilocarpine model of temporal lobe epilepsy by inhibiting nav1.6 sodium currents[J].Neurochem Res,2017,42(2):360- 374
[22] 夏烈新,陈立铭,彭晓艳,等.天麻素对癫痫大鼠海马神经元IL1、TNF及IL10的影响[J].临床神经病学杂志,2017,30(3):209- 213.
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