b FGF对颅脑损伤大鼠脑水肿、神经功能损伤及自噬相关蛋白表达的影响
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摘要
目的:研究碱性成纤维细胞生长因子(b FGF)对颅脑损伤(CI)大鼠脑水肿、神经功能损伤及自噬相关蛋白表达影响。方法:采用Feeney's自由落体硬膜外撞击法构建大鼠颅脑损伤模型,成年雄性SD大鼠随机分为对照(control)组、CI组及b FGF低、中和高剂量组,每组10只;对照组不给予硬膜外撞击,其它处理同模型组;b FGF低、中和高剂量组在建模成功30 min后分别给予2、4和6μg的b FGF腹腔注射,连续注射7 d。用改良神经功能评分法对大鼠神经功能进行评分。取大鼠脑组织,测定大鼠脑组织含水量,ELISA法测定脑组织中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和白细胞介素1β(IL-1β)含量,硫代巴比妥酸法测定丙二醛(MDA)含量,WST-8法检测超氧化物歧化酶(SOD)活性,比色法检测谷胱甘肽过氧化物酶(GSH-Px)活性,Western blot法测定脑组织中自噬相关蛋白LC3-Ⅱ和beclin-1的水平。结果:与control组相比,CI组神经功能评分明显升高(P <0.05);与CI组相比,b FGF低、中和高剂量组大鼠神经功能评分降低,脑组织含水量也降低(P <0.05),脑组织中TNF-α、IL-6和IL-1β水平降低(P <0.05),MDA含量降低(P <0.05),SOD和GSH-Px活性升高(P <0.05),LC3-Ⅱ和bec-lin-1蛋白水平降低(P <0.05)。结论:b FGF能够改善颅脑损伤大鼠神经功能,降低脑组织含水量,减少自噬相关蛋白LC3-Ⅱ和beclin-1的表达,这可能与其减轻炎症反应及氧化损伤有关。
AIM:To study the effects of basic fibroblast growth factor(b FGF)on brain edema,nerve function damage and autophagy related proteins in rats with head injury.METHODS:The rat model of craniocerebral injury(CI)was constructed.The rats were divided into control group,CI group,and low-,middle-and high-dose b FGF groups(n=10).The CI model was established in CI group,while the rats in control group were not given epidural impact.The rats in low-dose,middle-dose and high-dose b FGF groups were given b FGF at 2,4 and 6μg,respectively,by intraperitoneal injection after 30 min.The neurological function in the rats was evaluated by improved neurological function scoring.The rat brain tissues were taken,and the water content was detected.The levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and IL-1βin the brain tissue were measured by ELISA.The malondialdehyde(MDA)content was analyzed by thiobarbituric acid method.The activity of superoxide dismutase(SOD)was examined by WST-8 assay.The glutathine peroxidase(GSH-Px)activity was detected by colorimetric method.The protein levels of autophagy related proteins LC3-II and beclin-1 in the brain tissues were determined by Western blot.RESULTS:The neurological function score was increased significantly of the rats in CI group.The rat model of craniocerebral injury was successfully constructed.Neurological function scores in the rats in low-dose,middle-dose and high-dose b FGF groups were reduced,the water content of the brain tissue was also reduced(P<0.05).The levels of TNF-α,IL-6 and IL-1βwere decreased in the brain tissues(P<0.05),the content of MDA was declined(P<0.05),the activities of SOD and GSH-Px were increased(P<0.05),the protein levels of LC3-II and beclin-1 were decreased,compared with the untreated rats in CI group(P<0.05).CON-CLUSION:b FGF improves the nerve function of the rats with craniocerebral injury,reduces the water content of the brain tissue,reduces the expression of autophagic protein LC3-II and beclin-1.The mechanism is related to the inhibition of inflammatory reaction and oxidative damage.
AIM:To study the effects of basic fibroblast growth factor(b FGF)on brain edema,nerve function damage and autophagy related proteins in rats with head injury.METHODS:The rat model of craniocerebral injury(CI)was constructed.The rats were divided into control group,CI group,and low-,middle-and high-dose b FGF groups(n=10).The CI model was established in CI group,while the rats in control group were not given epidural impact.The rats in low-dose,middle-dose and high-dose b FGF groups were given b FGF at 2,4 and 6μg,respectively,by intraperitoneal injection after 30 min.The neurological function in the rats was evaluated by improved neurological function scoring.The rat brain tissues were taken,and the water content was detected.The levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and IL-1βin the brain tissue were measured by ELISA.The malondialdehyde(MDA)content was analyzed by thiobarbituric acid method.The activity of superoxide dismutase(SOD)was examined by WST-8 assay.The glutathine peroxidase(GSH-Px)activity was detected by colorimetric method.The protein levels of autophagy related proteins LC3-II and beclin-1 in the brain tissues were determined by Western blot.RESULTS:The neurological function score was increased significantly of the rats in CI group.The rat model of craniocerebral injury was successfully constructed.Neurological function scores in the rats in low-dose,middle-dose and high-dose b FGF groups were reduced,the water content of the brain tissue was also reduced(P<0.05).The levels of TNF-α,IL-6 and IL-1βwere decreased in the brain tissues(P<0.05),the content of MDA was declined(P<0.05),the activities of SOD and GSH-Px were increased(P<0.05),the protein levels of LC3-II and beclin-1 were decreased,compared with the untreated rats in CI group(P<0.05).CON-CLUSION:b FGF improves the nerve function of the rats with craniocerebral injury,reduces the water content of the brain tissue,reduces the expression of autophagic protein LC3-II and beclin-1.The mechanism is related to the inhibition of inflammatory reaction and oxidative damage.
引文
[1]Margulies S,Hicks R,Combination Therapies for Traumatic Brain Injury Workshop Leaders.Combination therapies for traumatic brain injury:prospective considerations[J].J Neurotrauma,2009,26(6):925-936.
[2]Simon DW,McGeachy MJ,Bayir H,et al.The far-reaching scope of neuroinflammation after traumatic brain injury[J].Nat Rev Neurol,2017,13(3):171-191.
[3]Dash PK,Mach SA,Moore AN.Enhanced neurogenesis in the rodent hippocampus following traumatic brain injury[J].J Neurosci Res,2015,63(4):313-319.
[4]Belli A,Sen J,Petzold A,et al.Metabolic failure precedes intracranial pressure rises in traumatic brain injury:a microdialysis study[J].Acta Neurochir,2008,150(5):461-469.
[5]Yang YH,Fu XB,Sun TZ,et al.b FGF and TGFbeta expression in rat kidneys after ischemic/reperfusional gut injury and its relationship with tissue repair[J].World JGastroenterol,2000,6(1):147-149.
[6]Wang ZG,Cheng Y,Yu XC,et al.b FGF protects against blood-brain barrier damage through junction protein regulation via PI3K-Akt-Rac1 pathway following traumatic brain injury[J].Mol Neurobiol,2015,53(10):7298-7311.
[7]魏冠.丙戊酸对大鼠创伤性颅脑损伤后炎症反应的影响[D].福州:福建医科大学,2016.
[8]Il’in DA,Arkhipov SA,Shkurupy VA.Analysis of IL-1α,b FGF,TGF-β1,IFNγ,MMP-1,and Cat D expression in multinuclea macrophages in vitro[J].Bull Exp Biol Med,2018,164(4):456-458.
[9]Ren ZC,Li RM,Yang YC,et al.Effect of hyperlipemia on b FGF expression in the hippocampus after cerebral ischemia-reperfusion injury[J].Chin J Neuroanatomy,2013,29(4):440-444.
[10]俞茜,杨彦玲,郝琴,等.b FGF对脊髓损伤的保护作用及其机制[J].延安大学学报(医学科学版),2016,14(1):60-63.
[11]向进,陈建良,陈洪,等.经颈总动脉推注rhbFGF治疗重症颅脑损伤的初步观察[J].中国临床神经外科杂志,2001,6(4):40-42.
[12]Petridis AK,Doukas A,Barth H,et al.Outcome of craniocerebral gunshot injuries in the civilian population.Prognostic factors and treatment options[J].Cen Eur Neurosurg,2010,72(1):5-14.
[13]Wang HF,Li WC,Xu N,et al.Transoral penetrating craniocerebral injury by a bamboo chopstick in a child[J].J Clin Neurosci,2013,20(5):746-748.
[14]Panourias IG,Slatinopoulos VK,Arvanitis DL.Penetrating craniocerebral injury caused by a pneumatic nail gun:an unsuccessful attempt of suicide[J].Clin Neurol Neurosurg,2006,108(5):490-492.
[15]Yang S,Zhao M,Han A,et al.Treatment of singultus following craniocerebral injury intranasal cavity drip infusion versus intramuscular injection of aminazine[J].Neural Regen Res,2007,2(10):621-624.
[16]赵永华,杨开敏,贾秀艳,等.丙氨酰谷氨酰胺对重型颅脑损伤患者肠黏膜通透性及血浆二胺氧化酶水平的影响[J].中国全科医学,2014,17(2):214-216.
[17]李茂林,王祝峰,章薇,等.红景天苷对大鼠颅脑损伤的保护作用研究[J].中华神经外科疾病研究杂志,2016,15(2):128-131.
[18]毛挺挺,方红波,王晓慧,等.碱性成纤维生长因子对大鼠成肌细胞氧化应激损伤的作用及其机制[J].中国应用生理学杂志,2017,33(2):159-163.
[19]樊凌云.碱性成纤维细胞生长因子对阿霉素致大鼠心肌氧化应激的影响[D].南宁:广西医科大学,2010.
[20]Fukui K,Takatsu H,Shinkai T,et al.Appearance of amyloid beta-like substances and delayed-type apoptosis in rat hippocampus CA1 region through aging and oxidative stress[J].J Alzheimers Dis,2005,8(3):299-309.
[21]Shu X,Chen F,Peng Q,et al.Potential role of autophagy in T-cell survival in polymyositis and dermatomyositis[J].Mol Med Rep,2017,16(2):1180-1188.
[22]Schlfli AM,Adams O,Galván JA,et al.Prognostic value of the autophagy markers LC3 and p62/SQSTM1 in early-stage non-small cell lung cancer[J].Oncotarget,2016,7(26):39544-39555.
[23]Chen Y,Li X,Wu X,et al.Autophagy-related proteins LC3 and Beclin-1 impact the efficacy of chemoradiation on esophageal squamous cell carcinoma[J].Pathol Res Pract,2013,209(9):562-567.
[24]Lu N,Wang B,Deng X,et al.Autophagy occurs within an hour of adenosine triphosphate treatment after nerve cell damage:the neuroprotective effects of adenosine triphosphate against apoptosis[J].Neural Regen Res,2014,9(17):1599-1605.
[2]Simon DW,McGeachy MJ,Bayir H,et al.The far-reaching scope of neuroinflammation after traumatic brain injury[J].Nat Rev Neurol,2017,13(3):171-191.
[3]Dash PK,Mach SA,Moore AN.Enhanced neurogenesis in the rodent hippocampus following traumatic brain injury[J].J Neurosci Res,2015,63(4):313-319.
[4]Belli A,Sen J,Petzold A,et al.Metabolic failure precedes intracranial pressure rises in traumatic brain injury:a microdialysis study[J].Acta Neurochir,2008,150(5):461-469.
[5]Yang YH,Fu XB,Sun TZ,et al.b FGF and TGFbeta expression in rat kidneys after ischemic/reperfusional gut injury and its relationship with tissue repair[J].World JGastroenterol,2000,6(1):147-149.
[6]Wang ZG,Cheng Y,Yu XC,et al.b FGF protects against blood-brain barrier damage through junction protein regulation via PI3K-Akt-Rac1 pathway following traumatic brain injury[J].Mol Neurobiol,2015,53(10):7298-7311.
[7]魏冠.丙戊酸对大鼠创伤性颅脑损伤后炎症反应的影响[D].福州:福建医科大学,2016.
[8]Il’in DA,Arkhipov SA,Shkurupy VA.Analysis of IL-1α,b FGF,TGF-β1,IFNγ,MMP-1,and Cat D expression in multinuclea macrophages in vitro[J].Bull Exp Biol Med,2018,164(4):456-458.
[9]Ren ZC,Li RM,Yang YC,et al.Effect of hyperlipemia on b FGF expression in the hippocampus after cerebral ischemia-reperfusion injury[J].Chin J Neuroanatomy,2013,29(4):440-444.
[10]俞茜,杨彦玲,郝琴,等.b FGF对脊髓损伤的保护作用及其机制[J].延安大学学报(医学科学版),2016,14(1):60-63.
[11]向进,陈建良,陈洪,等.经颈总动脉推注rhbFGF治疗重症颅脑损伤的初步观察[J].中国临床神经外科杂志,2001,6(4):40-42.
[12]Petridis AK,Doukas A,Barth H,et al.Outcome of craniocerebral gunshot injuries in the civilian population.Prognostic factors and treatment options[J].Cen Eur Neurosurg,2010,72(1):5-14.
[13]Wang HF,Li WC,Xu N,et al.Transoral penetrating craniocerebral injury by a bamboo chopstick in a child[J].J Clin Neurosci,2013,20(5):746-748.
[14]Panourias IG,Slatinopoulos VK,Arvanitis DL.Penetrating craniocerebral injury caused by a pneumatic nail gun:an unsuccessful attempt of suicide[J].Clin Neurol Neurosurg,2006,108(5):490-492.
[15]Yang S,Zhao M,Han A,et al.Treatment of singultus following craniocerebral injury intranasal cavity drip infusion versus intramuscular injection of aminazine[J].Neural Regen Res,2007,2(10):621-624.
[16]赵永华,杨开敏,贾秀艳,等.丙氨酰谷氨酰胺对重型颅脑损伤患者肠黏膜通透性及血浆二胺氧化酶水平的影响[J].中国全科医学,2014,17(2):214-216.
[17]李茂林,王祝峰,章薇,等.红景天苷对大鼠颅脑损伤的保护作用研究[J].中华神经外科疾病研究杂志,2016,15(2):128-131.
[18]毛挺挺,方红波,王晓慧,等.碱性成纤维生长因子对大鼠成肌细胞氧化应激损伤的作用及其机制[J].中国应用生理学杂志,2017,33(2):159-163.
[19]樊凌云.碱性成纤维细胞生长因子对阿霉素致大鼠心肌氧化应激的影响[D].南宁:广西医科大学,2010.
[20]Fukui K,Takatsu H,Shinkai T,et al.Appearance of amyloid beta-like substances and delayed-type apoptosis in rat hippocampus CA1 region through aging and oxidative stress[J].J Alzheimers Dis,2005,8(3):299-309.
[21]Shu X,Chen F,Peng Q,et al.Potential role of autophagy in T-cell survival in polymyositis and dermatomyositis[J].Mol Med Rep,2017,16(2):1180-1188.
[22]Schlfli AM,Adams O,Galván JA,et al.Prognostic value of the autophagy markers LC3 and p62/SQSTM1 in early-stage non-small cell lung cancer[J].Oncotarget,2016,7(26):39544-39555.
[23]Chen Y,Li X,Wu X,et al.Autophagy-related proteins LC3 and Beclin-1 impact the efficacy of chemoradiation on esophageal squamous cell carcinoma[J].Pathol Res Pract,2013,209(9):562-567.
[24]Lu N,Wang B,Deng X,et al.Autophagy occurs within an hour of adenosine triphosphate treatment after nerve cell damage:the neuroprotective effects of adenosine triphosphate against apoptosis[J].Neural Regen Res,2014,9(17):1599-1605.