【摘要】：背景：既往研究證實(shí)腦部炎癥反應(yīng)參與了癲癇的發(fā)生、發(fā)展,與癲癇所致腦組織損傷密切相關(guān)。藥物轉(zhuǎn)運(yùn)體P糖蛋白(P-glycoprotein, P-gp)被認(rèn)為是導(dǎo)致耐藥性癲癇(drug-resistant epilepsy, DRE)的重要機(jī)制之一,亦與炎性轉(zhuǎn)錄因子核因子κB (nuclear factor kappa B, NF-1κB)活化有關(guān)。新近研究發(fā)現(xiàn)癲癇腦內(nèi)可產(chǎn)生大量高遷移率族蛋白B1(high mobility group box1, HMGB1), HMGB1作為損傷相關(guān)分子模式成員之一,作用于模式識(shí)別受體介導(dǎo)免疫炎癥反應(yīng),調(diào)控NF-κB活性,我們前期研究發(fā)現(xiàn)癲癇腦內(nèi)P-gp過表達(dá)與NF-κB活化有關(guān),抑制NF-κB活性可下調(diào)P-gp表達(dá),那么,HMGB1能否調(diào)控P-gp蛋白的表達(dá)? 目的：觀察拮抗HMGB1對癲癇大鼠癲癇樣發(fā)作、神經(jīng)損傷、星型膠質(zhì)細(xì)胞活化的影響,探討HMGB1是否能通過活化NF-κB調(diào)控癲癇大鼠腦P-gp表達(dá)。 方法：將雄性SD大鼠分為假手術(shù)組(sham組,n=5),癲癇模型組(EP組,n=8),1400ng BoxA(HIMGB1拮抗劑)干預(yù)組(B1400組,n=8),140ng BoxA干預(yù)組(B140組,n=8),14ng BoxA干預(yù)組(B14組,n=8)。側(cè)腦室注射BoxA進(jìn)行干預(yù),sham組和EP組僅注射NS; BoxA干預(yù)后15min向海馬注射海人酸(kainic acid, KA)制作癲癇模型,sham組僅注射NS。記錄各組大鼠達(dá)到Ⅲ級癲癇發(fā)作的時(shí)間(seizure onset time, SOT)評價(jià)癲癇易感性。造模后24h取大鼠腦組織,H.E.染色大體觀察腦組織損傷,神經(jīng)元核抗原(neuronal nuclei, NeuN)染色評價(jià)神經(jīng)損傷程度,膠質(zhì)纖維酸性蛋白(glial fibrillary acidic protein, GFAP)染色評價(jià)星形膠質(zhì)細(xì)胞活化,HMGB1、p-NF-KB-p65、P-gp染色觀察BoxA干預(yù)對他們表達(dá)水平的影響。 主要結(jié)果： (1)各BoxA干預(yù)組SOT均較EP組延長(P0.05)； (2)各BoxA干預(yù)組神經(jīng)元損傷、丟失程度較EP組輕(P0.05)； (3)EP組星形膠質(zhì)細(xì)胞腫脹,各BoxA干預(yù)組GFAP表達(dá)少于EP組(P0.05)： (4)5組之間總體比較,HMGB1表達(dá)水平差異無統(tǒng)計(jì)學(xué)意義。EP組海馬神經(jīng)元未見HMGB1表達(dá),而膠質(zhì)細(xì)胞、內(nèi)皮細(xì)胞等HMMGB1表達(dá)增加；BoxA干預(yù)組神經(jīng)元有不同程度HMGB1表達(dá),而膠質(zhì)細(xì)胞、內(nèi)皮細(xì)胞HMGB1表達(dá)少于EP組； (5)各BoxA干預(yù)組P-gp表達(dá)少于EP組(P0.05)。 (6)B1400組、B140組p-NF-κB-p65表達(dá)少于EP組(P0.05),B14組表達(dá)有少于EP組的趨勢,但差異未顯示統(tǒng)計(jì)學(xué)意義； 結(jié)論：HMGB1拮抗劑BoxA可：降低大鼠的癲癇易感性；減輕神經(jīng)元損傷,減輕星形膠質(zhì)細(xì)胞腫脹、活化,具有神經(jīng)保護(hù)作用;抑制NF-KB活化,降低P-gP表達(dá)。結(jié)合我們既往的研究,BoxA可能通過抑制HMGB1,減少其誘導(dǎo)的NF-κB活化及下游P-gp表達(dá)。
[Abstract]:Background: previous studies have confirmed that brain inflammation is involved in the occurrence and development of epilepsy, and is closely related to brain tissue damage caused by epilepsy. Drug transporter P-glycoprotein (P-gp) is considered to be one of the important mechanisms leading to drug-resistant epilepsy (drug-resistant epilepsy, DRE), and also related to the activation of inflammatory transcription factor 魏 B (nuclear factor kappa B, NF-1 魏 B. Recently, it has been found that a large number of high mobility group protein B1 (high mobility group box1, HMGB1 can be produced in the epileptic brain. As one of the damage related molecular model members, HMGB1 acts as a pattern recognition receptor mediating immune inflammation and regulating the activity of NF- 魏 B. Our previous study found that the overexpression of P-gp in the epileptic brain is related to the activation of NF- 魏 B, and the inhibition of the activity of NF- 魏 B can down-regulate the expression of P-gp. Can HMGB1 regulate the expression of P-gp protein? Aim: to observe the effects of antagonistic HMGB1 on epileptoid seizure, nerve injury and astrocyte activation in epileptic rats, and to investigate whether HMGB1 can regulate the expression of P-gp in the brain of epileptic rats by activating NF- 魏 B. Methods: male SD rats were divided into three groups: sham operation group (sham group, nong5), epileptic model group (EP group, nong8), 1400ng BoxA (HIMGB1 antagonist) intervention group (B1400, nong8), 140ng BoxA intervention group (B140, nong8), 14ng BoxA intervention group (B14, N8). Intracerebroventricular injection of BoxA was performed in sham group and EP group. After NS; BoxA intervention, 15min injected kainic acid (kainic acid, KA) into hippocampus to make epileptic model, sham group only injected NS.. The time of reaching grade 鈪，

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