【摘要】：癲癇是一種慢性反復(fù)發(fā)作的嚴重的神經(jīng)系統(tǒng)常見疾病之一。盡管目前各種各樣的抗癲癇藥物在臨床上使用,但由于其發(fā)病機理尚不清楚,仍有20-30%的癲癇患者癥狀得不到有效控制。因此,對癲癇發(fā)病機制的深入研究,有益于指導癲癇的預(yù)防和治療。越來越多的研究表明異常的神經(jīng)網(wǎng)絡(luò)在癲癇發(fā)病中起著重要作用。屬于酪氨酸家族成員的EphB/ephrinB系統(tǒng),是一類神經(jīng)軸突導向因子,他們相互作用在中樞神經(jīng)系統(tǒng)內(nèi)神經(jīng)元軸突導向、特異性突觸連接、神經(jīng)元樹突結(jié)構(gòu)等方面具有非常重要作用。本課題主要研究EphB/ephrinB各亞型在臨床難治性顳葉癲癇患者和癲癇大鼠模型中的表達,以求找出可能與癲癇形成有關(guān)的亞型;并在無鎂誘導的離體海馬腦片癲癇模型和兩種經(jīng)典的慢性癲癇動物模型上,應(yīng)用功能性重組融合蛋白和慢病毒介導的基因轉(zhuǎn)染技術(shù)進行干預(yù),通過電生理學、分子生物學、動物行為學以及形態(tài)學檢測,探討其在癲癇發(fā)作和形成過程中的作用和可能機制,為癲癇發(fā)病機制提供新思路和新途徑,以及為臨床上尋找抗癲癇治療新靶點和新的抗癲癇藥物研發(fā)提供實驗依據(jù)。第一部分EphB/ephrinB在難治性顳葉癲癇患者及大鼠癲癇模型中的表達研究目的:檢測EphB/ephrinB各亞型在臨床難治性顳葉癲癇患者和氯化鋰-匹羅卡品誘導的癲癇大鼠腦組織中的表達,以求找出可能與癲癇形成有關(guān)的EphB/ephrinB亞型。方法:1.從第三軍醫(yī)大學附屬第二醫(yī)院(新橋醫(yī)院)神經(jīng)外科腦標本庫中隨機收集20例難治性顳葉癲癇患者和20例嚴重顱腦外傷需要去骨瓣減壓手術(shù)治療的對照患者手術(shù)后顳葉皮層標本。2.SD成年雄性大鼠隨機分成正常對照組和癲癇模型組(每組10只),模型組給與氯化鋰-匹羅卡品腹腔注射誘導癲癇模型。3.用RT-qPCR檢測EphB/ephrinB各亞型在臨床標本和動物模型上的表達,篩選出mRNA表達一致的亞型,然后用Westernblot和免疫組織化學染色驗證其蛋白表達,接著進一步用Westernblot檢測其功能性磷酸化蛋白水平,并用免疫熒光雙標對其細胞表達進行定位。結(jié)果:1.在癲癇患者和癲癇大鼠腦組織中EphB3和ephrinB3mRNA表達一致性升高(p0.05),而EphBl/2/4/6和ephrinl/2mRNA表達趨勢不一致。2.Westernblot和免疫組織化學染色證實EphB3和ephrinB3蛋白表達在癲癇標本中一致性升高(p0.05)。3.功能性p-EphB3蛋白表達在癲癇標本中升高(p0.05),而p-ephrinB蛋白表達在癲癇標本中沒有改變(p0.05)。4.免疫熒光雙標提示EphB3在癲癇標本中主要表達于神經(jīng)元胞質(zhì)和胞膜。結(jié)論:EphB3和p-EphB3在難治性顳葉癲癇患者和氯化鋰-匹羅卡品誘導的癲癇大鼠腦組織中表達升高,提示EphB3可能與癲癇的形成有關(guān)。第二部分EphB3功能改變對大鼠海馬腦片神經(jīng)元興奮性的影響目的:為了探討EphB3對癲癇發(fā)作的影響,我們在無鎂誘導的離體海馬腦片癲癇模型上,用全細胞膜片鉗檢測EphB3功能改變對海馬神經(jīng)元興奮性的影響。方法:健康雄性SD大鼠麻醉后直接斷頭取腦制作海馬腦片,在正常人工腦脊液灌流基礎(chǔ)上,換成無鎂人工腦脊液誘導離體腦片癲癇模型,然后將EphB3活性調(diào)節(jié)劑功能性重組融合蛋白(抑制劑EphB3-Fc、激活劑ephrinB3-Fc)和對照Fc加入灌流液中進行干預(yù),用全細胞膜片鉗檢測各干預(yù)前后神經(jīng)元動作電位頻率。結(jié)果:無鎂人工腦脊液可明顯升高神經(jīng)元動作電位頻率(p0.05);EphB3-Fc(10nm和100nm)可降低無鎂誘導的神經(jīng)元動作電位頻率(p0.05);ephrinB3-Fc(10nm和100nm)可升高無鎂誘導的神經(jīng)元動作電位頻率(p0.05);而Fc(10nm和100nm)對無鎂誘導的神經(jīng)元動作電位頻率沒有影響(p0.05)。融合蛋白(Fc、EphB3-Fc和ephrinB3-Fc10nm)對正常人工腦脊液環(huán)境下的神經(jīng)元動作電位頻率沒有影響(p0.05)。結(jié)論:無鎂人工腦脊液可誘導離體海馬腦片癲癇模型。抑制EphB3活性可降低無鎂誘導的神經(jīng)元興奮性,激活EphB3活性可升高無鎂誘導的神經(jīng)元興奮性,提示EphB3功能改變可影響癲癇的發(fā)作。第三部分EphB3功能改變對癲癇大鼠行為學及下游相關(guān)RhoGEF基因表達的影響目的:為了進一步明確EphB3對癲癇發(fā)作和形成的影響,我們在兩種經(jīng)典慢性癲癇動物模型上,觀察EphB3功能改變對大鼠行為學、海馬齒狀回區(qū)苔蘚纖維出芽、內(nèi)分子層樹突棘密度和EphB3下游相關(guān)RhoGEF基因表達的影響。方法:健康雄性SD大鼠側(cè)腦室置管后恢復(fù)一周,隨機分成單純功能性重組融合蛋白干預(yù)、氯化鋰-匹羅卡品誘導癲癇模型和戊四氮點燃癲癇模型三個大組。單純功能性重組融合蛋白干預(yù)組大鼠又隨機分成ephrinB3-Fc組、EphB3-Fc組、對照Fc組和正常對照(PBS)組(每組5只),分別用ephrinB3-Fc、EphB3-Fc、Fc和PBS通過側(cè)腦室置管注射連續(xù)干預(yù)一周,于干預(yù)后第7、14、30和60天麻醉后斷頭取腦,用Westernblot檢測各組海馬組織中EphB3和p-EphB3蛋白表達。癲癇模型組大鼠又隨機分成4個亞組,即模型對照(PBS)組、模型+對照Fc組、模型+ephrinB3-Fc組和模型+EphB3-Fc組(每組10只),通過側(cè)腦室置管分別注射等量無菌PBS和融合蛋白(Fc、ephrinB3-Fc和EphB3-Fc)連續(xù)干預(yù)一周后造模。氯化鋰-匹羅卡品模型觀察大鼠急性發(fā)作程度、急性潛伏期和慢性期自發(fā)性發(fā)作次數(shù),戊四氮模型組觀察大鼠每天發(fā)作程度和完全被點燃的時間。應(yīng)用Timm染色檢測癲癇大鼠海馬齒狀回苔蘚纖維出芽情況,高爾基染色檢測癲癇大鼠海馬齒狀回內(nèi)分子層區(qū)樹突棘密度,RT-qPCR和Westernblot檢測癲癇大鼠海馬組織中EphB3下游相關(guān)RhoGEF基因(Kalirin、Intersectin1和Intersectin2)表達情況。結(jié)果:1.EphB3-Fc可長時間(7-60d)降低大鼠海馬組織p-EphB3蛋白表達(p0.05),ephrinB3-F可長時間(7-60d)增加大鼠海馬組織p-EphB3蛋白表達(p0.05),而Fc對大鼠海馬組織p-EphB3蛋白表達沒有影響(p0.05)。2.EphB3-Fc可降低氯化鋰-匹羅卡品癲癇大鼠的急性發(fā)作級別、延長急性潛伏期和減少慢性期自發(fā)性發(fā)作次數(shù)(p0.05),ephrinB3-Fc則可加重氯化鋰-匹羅卡品癲癇大鼠的急性發(fā)作級別、縮短急性潛伏期和增加慢性期自發(fā)性發(fā)作次數(shù)(p0.05)。同時EphB3-Fc可降低戊四氮癲癇大鼠的發(fā)作級別(11-33d)和延長發(fā)作潛伏期(p0.05),ephrinB3-Fc則可加重戊四氮癲癇大鼠的發(fā)作級別(9-29d)和縮短發(fā)作潛伏期(p0.05),Fc對氯化鋰-匹羅卡品和戊四氮癲癇大鼠的上述行為學沒有影響(p0.05)。3.EphB3-Fc可減輕癲癇大鼠海馬齒狀回區(qū)苔蘚纖維出芽程度和降低內(nèi)分子層樹突棘密度,ephrinB3-Fc則加重癲癇大鼠海馬齒狀回區(qū)苔蘚纖維出芽程度和增加內(nèi)分子層樹突棘密(p0.05),Fc對癲癇大鼠海馬齒狀回區(qū)苔蘚纖維出芽程度和內(nèi)分子層樹突棘密沒有影響(p0.05)。4.功能性重組融合蛋白對癲癇大鼠海馬組織Intersectin1和Intersectin2mRNA表達沒有影響(p0.05)。EphB3-Fc可降低癲癇大鼠海馬組織中KalirinmRNA和蛋白表達,ephrinB3-Fc則增加癲癇大鼠海馬組織KalirinmRNA和蛋白表達(p0.05),而Fc對癲癇大鼠海馬組織KalirinmRNA和蛋白表達沒有影響(p0.05)。結(jié)論:1.功能性重組融合蛋白(ephrinB3-Fc和EphB3-Fc)可長時間(7-60d)影響EphB3功能。2.EphB3功能改變可影響癲癇大鼠急性發(fā)作程度、潛伏期長短以及慢性期自發(fā)性發(fā)作頻率。3.EphB3功能改變可調(diào)節(jié)癲癇大鼠海馬區(qū)苔蘚纖維出芽和內(nèi)分子層樹突棘密度。4.Kalirin可能為EphB3潛在的下游作用靶點。第四部分EphB3通過Kalirin參與癲癇的發(fā)作與形成目的:為了探索EphB3在癲癇發(fā)作和形成中的可能機制,我們在兩種經(jīng)典慢性癲癇動物模型上,用慢病毒介導Kalirin-shRNA沉默Kalirin基因后,觀察激活EphB3功能對大鼠行為學、海馬齒狀回區(qū)苔蘚纖維出芽、內(nèi)分子層樹突棘密度的影響,以明確EphB3是否是通過Kalirin參與癲癇的發(fā)作與形成。方法:用Westernblot檢測難治性顳葉癲癇患者和癲癇大鼠腦組織中Kalirin蛋白表達。健康雄性SD大鼠隨機分成Control、LV-GFP和LV-Kalirin-shRNA三組,分別通過側(cè)腦室置管注射等量生理鹽水、LV-GFP和LV-Kalirin-shRNA。一部分LV-Kalirin-shRNA干預(yù)組大鼠分別于注射后第3、7、14、30和60天(7天組大鼠10只,其余每組5只)麻醉后斷頭取腦,熒光顯微鏡下觀察大鼠海馬以及皮層綠色熒光表達,Westernblot檢測海馬Kalirin蛋白表達。另一部分大鼠(30只)于側(cè)腦室置管注射生理鹽水和慢病毒后一周,用氯化鋰-匹羅卡品和戊四氮制備癲癇模型(每組大鼠5只)。觀察各組大鼠行為學改變,Westernblot檢測癲癇大鼠海馬組織中EphB3和p-EphB3蛋白表達情況。剩余部分大鼠(30只)于側(cè)腦室置管注射生理鹽水和慢病毒一周后,繼續(xù)通過側(cè)腦室置管注射ephrinB3-Fc連續(xù)干預(yù)一周,然后用氯化鋰-匹羅卡品和戊四氮制備癲癇模型(每組大鼠5只)。觀察各組大鼠行為學改變、海馬齒狀回苔蘚纖維出芽和內(nèi)分子層區(qū)樹突棘密度情況。結(jié)果:1.在難治性顳葉癲癇患者和癲癇大鼠腦組織中Kalirin蛋白表達升高(p0.05)。2.熒光顯微鏡下觀察到慢病毒綠色熒光廣泛分布于大鼠大腦皮層和海馬區(qū)。LV-Kalirin-shRNA從注射后第7天開始有效抑制Kalirin蛋白表達,并持續(xù)至注射后的第60天(p0.05)。3.LV-Kalirin-shRNA降低氯化鋰-匹羅卡品癲癇大鼠的急性發(fā)作級別、延長急性潛伏期和減少慢性期自發(fā)性發(fā)作次數(shù)(p0.05),同時也降低戊四氮癲癇大鼠的發(fā)作級別(9-35d)和延長發(fā)作潛伏期(p0.05)。LV-Kalirin-shRNA對癲癇大鼠海馬組織EphB3和p-EphB3蛋白表達沒有影響(p0.05)。4.沉默內(nèi)源性的Kalirin能降低ephrinB3-Fc所引起的氯化鋰-匹羅卡品癲癇大鼠的急性發(fā)作級別、急性潛伏期、慢性期自發(fā)性發(fā)作次數(shù)以及戊四氮癲癇大鼠的發(fā)作級別(11-29d)和發(fā)作潛伏期(p0.05)。5.沉默內(nèi)源性的Kalirin能減輕和降低ephrinB3-Fc所引起的癲癇大鼠海馬齒狀回區(qū)苔蘚纖維出芽程度和內(nèi)分子層樹突棘密度(p0.05)。結(jié)論:1.Kalirin在難治性TLE患者和癲癇大鼠腦組織中表達升高。2.沉默內(nèi)源性Kalirin可減輕癲癇大鼠發(fā)作程度、延長潛伏期和減少慢性期大鼠自發(fā)性發(fā)作頻率。3.沉默內(nèi)源性Kalirin可逆轉(zhuǎn)EphB3激活所引起的癲癇大鼠行為學和海馬神經(jīng)環(huán)路形態(tài)學改變。4.EphB3通過對RhoGEFKalirin的調(diào)節(jié)影響海馬神經(jīng)環(huán)路重建參與癲癇的發(fā)作和形成。
[Abstract]:Epilepsy is one of the most common chronic recurrent neurological diseases. Although various antiepileptic drugs are used in clinic, there are still 20-30% epileptic patients whose symptoms can not be effectively controlled because of the unclear pathogenesis. Therefore, in-depth study on the pathogenesis of epilepsy is helpful to guide epilepsy. Prevention and treatment. More and more studies have shown that abnormal neural networks play an important role in epilepsy. EphB/ephrinB system, a member of tyrosine family, is a class of neuronal axon-directing factors that interact with neuronal axon-directing, specific synaptic connections, and neuronal dendritic structures in the central nervous system. Masks play an important role in the development of intractable temporal lobe epilepsy and epilepsy in rats. EphB/ephrinB subtypes were used to identify potential epilepsy-related subtypes, and in magnesium-free hippocampal slice epilepsy model and two classical chronic epilepsy animal models. Sexual recombinant fusion protein and lentivirus-mediated gene transfection were used to intervene in the process of epileptic seizure and its possible mechanism by electrophysiology, molecular biology, animal behavior and morphology detection, which provided new ideas and new ways for epileptic pathogenesis and clinical anti-epileptic treatment. Objective: To detect the expression of EphB/ephrinB subtypes in the brain of patients with refractory temporal lobe epilepsy and epileptic rats induced by lithium-pilocarpine. Methods: 1. Twenty patients with refractory temporal lobe epilepsy and 20 control patients with severe craniocerebral trauma requiring decompressive craniotomy were randomly collected from the neurosurgical brain specimen bank of the Second Affiliated Hospital of the Third Military Medical University (Xinqiao Hospital) and compared with the control group. 2. SD Male rats were randomly divided into normal control group and epilepsy model group (10 rats in each group). The model group was given lithium chloride-pilocarpine intraperitoneally to induce epilepsy model. 3. The expression of EphB/ephrinB subtypes in clinical specimens and animal models was detected by RT-qPCR, and the mRNA subtypes with the same expression were screened out. The expression of ephB3 and ephrinB3 mRNA in the brain tissues of epileptic patients and epileptic rats was consistent with that of epileptic rats (p0.05), but the expression of EphBl/2/4/6 and ephrinl/2 mRNA tended to increase. Western blot and immunohistochemical staining confirmed that EphB3 and ephrinB3 protein expression in epileptic specimens increased consistently (p0.05). 3. Functional p-EphB3 protein expression in epileptic specimens increased (p0.05), while p-ephrinB protein expression in epileptic specimens did not change (p0.05). 4. Immunofluorescence double labeling suggested that EphB3 was present in epileptic specimens. EphB3 and p-EphB3 were mainly expressed in the cytoplasm and membrane of neurons. Conclusion: EphB3 and p-EphB3 were elevated in the brain tissues of refractory temporal lobe epilepsy patients and lithium-pilocarpine-induced epilepsy rats, suggesting that EphB3 may be related to the formation of epilepsy. To investigate the effect of EphB3 on seizures, we used whole-cell patch clamp technique to detect the effect of EphB3 on the excitability of hippocampal neurons in a magnesium-free hippocampal slice epilepsy model. The epilepsy model was induced by artificial cerebrospinal fluid (ACF), and then the functional recombinant fusion protein (inhibitor EphB3-Fc, activator ephrinB3-Fc) and the control Fc were added into the perfusion fluid for intervention. The action potential frequencies of neurons before and after intervention were detected by whole-cell patch clamp. Neuronal action potential frequency (p0.05); EphB3-Fc (10 nm and 100 nm) decreased Mg-free neuronal action potential frequency (p0.05); ephrinB3-Fc (10 nm and 100 nm) increased Mg-free neuronal action potential frequency (p0.05); Fc (10 nm and 100 nm) had no effect on Mg-free neuronal action potential frequency (p0.05). Fusion protein (Fc) EphB3-Fc and ephrinB3-Fc10nm had no effect on neuronal action potential frequencies in normal artificial cerebrospinal fluid environment (p0.05). Conclusion: Magnesium-free artificial cerebrospinal fluid can induce epilepsy model in isolated hippocampal slices. It is suggested that the changes of EphB3 function may influence the onset of epilepsy. Part III The effect of EphB3 function on the behavior and downstream rhoGEF gene expression in epileptic rats Objective: To further clarify the effect of EphB3 on seizures and formation, we observed the behavior of epileptic rats in two classical chronic epileptic animal models. Methods: Healthy male SD rats were randomly divided into three groups: single functional recombinant fusion protein intervention, lithium chloride-pilocarpine induced epilepsy model and pentylenetetrazol kindled epilepsy model. Rats in the simple functional recombinant fusion protein intervention group were randomly divided into ephrinB3-Fc group, EphB3-Fc group, control Fc group and normal control (PBS) group (5 rats in each group). EphrinB3-Fc, EphB3-Fc, Fc and PBS were injected into the lateral ventricle for one week. Brains were cut off at the 7th, 14th, 30th and 60th days after the intervention, and brain was taken out by Western blot. EphB3 and p-EphB3 protein expression were detected in the hippocampus of each group. Epileptic model rats were randomly divided into four subgroups: model control group (PBS), model + control group (Fc), model + ephrinB3-Fc and model + EphB3-Fc (10 rats in each group). The same amount of aseptic PBS and fusion protein (Fc, ephrinB3-Fc and EphB3-Fc) were injected into the lateral ventricle of rats respectively. Lithium chloride-pilocarpine model was used to observe the degree of acute attack, the number of spontaneous attacks in acute latency and chronic phase, and pentylenetetrazol model group was used to observe the degree of attack and the time of complete kindling. The dendritic spine density in the molecular layer of the dentate gyrus of the hippocampus of epileptic rats was measured. The expression of RhoGEF gene (Kalirin, Intersectin 1 and Intersectin2) downstream of EphB3 in the hippocampus of epileptic rats was detected by RT-qPCR and Western blot. Results: 1. EphB3-Fc could decrease the expression of p-EphB3 protein (p0.05) for a long time (7-60 days) in the hippocampus of epileptic rats. Time (7-60 days) increased the expression of p-EphB3 protein in rat hippocampus (p0.05), but Fc had no effect on the expression of p-EphB3 protein in rat hippocampus (p0.05). 2. EphB3-Fc could decrease the acute seizure grade, prolong the acute latency and reduce the number of spontaneous seizures in chronic phase (p0.05). EphrinB3-Fc could aggravate chlorine content in rat hippocampus. EphB3-Fc could decrease the onset grade (11-33 days) and prolong the onset latency (p0.05). EphrinB3-Fc could aggravate the onset grade (9-29 days) and shorten the onset of pentylenetetrazol epilepsy in rats. During the incubation period (p0.05), Fc had no effect on the above behavior of lithium chloride-pilocarpine and pentylenetetrazol epileptic rats (p0.05). 3. EphB3-Fc could reduce the degree of mossy fiber sprouting in hippocampal dentate gyrus and the density of dendritic spines in inner molecular layer of epileptic rats, while ephrinB3-Fc aggravated the degree and increase of mossy fiber sprouting in hippocampal dentate gyrus of epileptic rats. Fc had no effect on the degree of mossy fiber sprouting and dendritic dendritic dendrite density in hippocampal dentate gyrus of epileptic rats (p0.05). 4. Functional recombinant fusion protein had no effect on the expression of Intersectin1 and Intersectin2 mRNA in hippocampus of epileptic rats (p0.05). EphB3-Fc could decrease the expression of K in hippocampus of epileptic rats. Alarin mRNA and protein expression, ephrin B3-Fc increased Kalirin mRNA and protein expression in hippocampus of epileptic rats (p0.05), but Fc had no effect on Kalirin mRNA and protein expression in hippocampus of epileptic rats (p0.05). Conclusion: 1. Functional recombinant fusion proteins (ephrin B3-Fc and EphB3-Fc) can affect the function of EphB3 for a long time (7-60 days). EphB3 can regulate mossy fiber budding and dendritic spine density in hippocampus of epileptic rats. 4. Kalirin may be a potential downstream target of EphB3. Part IV EphB3 participates in epileptic attack and shape through Kalirin. AIM: To explore the possible mechanism of EphB3 in epileptic seizures and epilepsy formation, we observed the effects of activating EphB3 on rat behavior, mossy fiber budding and dendritic spine density in hippocampal dentate gyrus after Kalirin-shRNA silencing in two classical chronic epileptic animal models. Methods: The expression of Kalirin protein in brain tissues of refractory temporal lobe epilepsy patients and epileptic rats was detected by Western blot. Healthy male SD rats were randomly divided into three groups: control, LV-GFP and LV-Kalirin-shRNA, which were injected with normal saline, LV-GFP and LV-Kalir through lateral ventricle catheterization respectively. In-shRNA. Some LV-Kalirin-shRNA-treated rats were anesthetized at 3,7,14,30 and 60 days after injection (10 rats in 7-day group and 5 rats in each group). The expression of green fluorescence in hippocampus and cortex was observed under fluorescence microscope. The expression of Kalirin protein in hippocampus was detected by Western blot. Another part of rats (30 rats) were placed cannula in lateral ventricle. Epilepsy models were established with lithium chloride-pilocarpine and pentylenetetrazol (5 rats in each group) one week after injection of normal saline and lentiviruses. Behavioral changes were observed in each group. Expression of EphB3 and p-EphB3 protein in hippocampus of epileptic rats was detected by Western blot. The remaining 30 rats (30 rats) were injected with normal saline and lentiviruses in lateral ventricles. After one week, the epileptic model was established by injecting ephrinB3-Fc into the lateral ventricle for one week, then lithium chloride-pilocarpine and pentylenetetrazol were used to prepare the epileptic model (5 rats in each group). Behavioral changes, mossy fiber sprouting in the dentate gyrus and dendritic spine density in the inner molecular layer of the hippocampus were observed. LV-Kalirin-shRNA effectively inhibited the expression of Kalirin protein from the 7th day after injection and lasted until the 60th day after injection (p0.05). 3. LV-Kalirin-shRNA reduced chlorination. LV-Kalirin-shRNA had no effect on the expression of EphB3 and p-EphB3 protein in hippocampus of epileptic rats (p0.05). The silencing of endogenous Kalirin can reduce the acute epileptic grade, acute latency, the number of spontaneous seizures in chronic phase, the onset grade (11-29 days) and the onset latency (p0.05) of epileptic rats induced by ephrinB3-Fc. The silencing of endogenous Kalirin can alleviate and reduce the induction of ephrinB3-Fc. The degree of mossy fiber sprouting and dendritic spine density of inner molecular layer in dentate gyrus of hippocampus in epileptic rats (p0.05). Conclusion: 1. Kalirin in refractory TLE
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：R742.1

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