本文選題：七氟醚　切入點(diǎn)：前額葉皮層　出處：《中國(guó)病理生理雜志》2015年10期 　論文類型：期刊論文

【摘要】：目的:探究七氟醚對(duì)新生大鼠前額葉皮層(prefrontal cortex,PFC)樹(shù)突發(fā)育的影響及與細(xì)胞周期依賴性蛋白激酶5(cyclin dependent kinase 5,Cdk5)-坍塌反應(yīng)調(diào)節(jié)蛋白(collapsin response mediator protein,CRMP)通路的關(guān)系。方法:88只出生后7 d的SD大鼠隨機(jī)分為4組,每組22只:空白對(duì)照組(Air+NS組),Cdk5抑制劑roscovitine(Ros)對(duì)照組(Air+Ros組),Sevo+NS組,Sevo+Ros組。前2組吸入空氣,后2組吸入2.8%七氟醚4 h。Air+Ros組和Sevo+Ros組在麻醉前15 min經(jīng)腹腔注射10 mg/kg的Cdk5抑制劑roscovitine。其余2組則在麻醉前15min經(jīng)腹腔注射150μL生理鹽水。麻醉結(jié)束后,每組各取5只幼鼠,取出新鮮皮質(zhì)組織,Western blot檢測(cè)P35、P25、Cdk5,CRMP1、2、4的蛋白表達(dá)以及CRMP2 Ser 522的磷酸化水平。剩余幼鼠繼續(xù)飼養(yǎng),出生后30 d每組各取6只幼鼠,取腦進(jìn)行高爾基染色,制備冰凍切片,鏡下觀察神經(jīng)元形態(tài)。其余幼鼠在出生后25~27 d進(jìn)行曠場(chǎng)試驗(yàn),31~32 d進(jìn)行情景依賴性恐懼記憶檢測(cè)。結(jié)果:(1)與Air+NS組比較,Sevo+NS組的P35減少了47.32%,同時(shí)P25增加了173.77%(P0.05);Sevo+Ros組的P35比Sevo+NS組增加了78.81%,其P25則減少了60.22%(P0.05)。Cdk5表達(dá)組間差異尚無(wú)統(tǒng)計(jì)學(xué)意義。(2)與Air+NS組比較,Sevo+NS組的總CRMP1、2和4表達(dá)分別下降了36.22%、53.74%和51.28%(P0.05),同時(shí)p-CRMP2 Ser522/CRMP2比值增加了96.20%(P0.05);Sevo+Ros組的總CRMP1和4與Sevo+NS組比較分別增加了56.95%和88.91%(P0.05),而pCRMP2 Ser522/CRMP2比值下降了43.58%(P0.05)。(3)與Air+NS組比較,Sevo+NS組的樹(shù)突總長(zhǎng)度、第二級(jí)樹(shù)突長(zhǎng)度以及60和80μm直徑圓環(huán)上的交點(diǎn)數(shù)均顯著下降(P0.01)。Sevo+Ros組的樹(shù)突總長(zhǎng)度、第二級(jí)樹(shù)突長(zhǎng)度以及60和80μm直徑圓環(huán)上的交點(diǎn)數(shù)均比Sevo+NS組顯著增加(P0.05)。(4)在恐懼記憶實(shí)驗(yàn)中,與Air+NS組比較,Sevo+NS組大鼠的"木僵"時(shí)間比Air+NS組減少約21%(P0.05)。Sevo+Ros組的"木僵"時(shí)間比Sevo+NS組顯著增加(P0.05)。各組幼鼠在曠場(chǎng)實(shí)驗(yàn)的結(jié)果均無(wú)統(tǒng)計(jì)學(xué)差異。結(jié)論:七氟醚抑制了新生大鼠前額葉皮層神經(jīng)元樹(shù)突的正常生長(zhǎng)發(fā)育和幼鼠學(xué)習(xí)記憶能力,而Cdk5-CRMP通路激活可能是七氟醚引起神經(jīng)發(fā)育毒性的機(jī)制之一。
[Abstract]:Objective: to investigate the effect of sevoflurane on dendritic development of prefrontal cortex in neonatal rats and its relationship with the cell cycle dependent protein kinase (5(cyclin dependent kinase 5) Cdk5- collapse regulatory protein (CRMP) pathway. D SD rats were randomly divided into 4 groups. There were 22 rats in each group: control group: air NS group: roscovitine 5 inhibitor; control group: air Ros group; Sevo Ros group; the first two groups inhaled air, the first two groups inhaled air, the first two groups inhaled air, and the first two groups inhaled air. The latter two groups inhaled 2.8% sevoflurane 4 Ros group and Sevo Ros group injected roscovitine, a Cdk5 inhibitor of 10 mg/kg, 15 min before anesthesia. The other two groups were injected with 150 渭 L saline intraperitoneally before anesthesia. After anesthesia, 5 young rats were taken from each group. Fresh cortical tissues were taken out to detect the protein expression of P35, P25, Cdk5 and CRMP1, and the phosphorylation level of CRMP2 Ser 522. The remaining young rats were reared and 6 young rats were collected from each group 30 days after birth, and their brains were stained with Golgi to prepare frozen sections. The morphology of neurons was observed under microscope. The other young rats were tested for situational dependent fear memory on day 31 and 32 after birth. Results: compared with Air NS group, P35 of Sevo NS group decreased by 47.32 and P25 increased by 173.77g and 173.77g / kg respectively compared with that of Sevo Ros group. Compared with Sevo NS group, P35 increased 78.81%, while P25 decreased 60.22%(P0.05).Cdk5 expression. There was no significant difference between them.) compared with Air NS group, total CRMP1m-2 and 4 expression in Sevo NS group decreased by 36.22% and 51.28%, respectively, and p-CRMP2 Ser522/CRMP2 ratio increased by 96.20% in Sevo Ros group. 4Compared with Sevo NS group by 56.95% and 88.91%, respectively, the total dendritic length of Air NS group was decreased by 43.58% and 43.58%, respectively, compared with that of Air NS group. The length of secondary dendrites and the number of intersection points on 60 and 80 渭 m diameter rings were significantly decreased. The total length of dendrites, the length of secondary dendrites and the number of intersection points on rings of 60 and 80 渭 m diameter in the P0.01Sevo Ros group were significantly increased than those in the Sevo NS group. Compared with the Air NS group, the "stiffness" time of the Sevo NS group was significantly lower than that of the Air NS group. The "stiff" time of the 21%(P0.05).Sevo Ros group was significantly higher than that of the Sevo NS group. There was no statistical difference in the results of the open field experiment among the young rats in each group. Conclusion: sevoflurane is not significantly different in all the young rats. It inhibited the normal growth and development of dendrites of prefrontal cortex neurons and the learning and memory ability of young rats. Activation of Cdk5-CRMP pathway may be one of the mechanisms of neurotoxicity induced by sevoflurane.
【作者單位】： 中山大學(xué)附屬孫逸仙紀(jì)念醫(yī)院麻醉科;佛山市禪城區(qū)中心醫(yī)院麻醉科;
【基金】：廣東省自然科學(xué)基金資助項(xiàng)目(No.S2013010016207) 廣東省科技社會(huì)發(fā)展項(xiàng)目(No.2014A020212147) 廣東省醫(yī)學(xué)科學(xué)技術(shù)研究基金(No.A2013207)
【分類號(hào)】：R614

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