海馬膠質(zhì)細(xì)胞活化在七氟醚誘發(fā)老年大鼠認(rèn)知功能損害中的作用
發(fā)布時(shí)間：2018-08-03 07:37
【摘要】：認(rèn)知功能障礙是麻醉手術(shù)后常見(jiàn)的神經(jīng)系統(tǒng)并發(fā)癥,已成為影響老年人術(shù)后恢復(fù)進(jìn)程和生活質(zhì)量的重大問(wèn)題[1]。七氟醚是臨床常用的吸入性麻醉藥,對(duì)術(shù)后認(rèn)知功能有影響[2],但發(fā)生機(jī)制尚不明確。因此深入了解七氟醚誘發(fā)術(shù)后認(rèn)知功能障礙的機(jī)理以及找到適宜七氟醚麻醉深度減少甚至消除認(rèn)知功能障礙的發(fā)生一直是麻醉學(xué)科研的熱點(diǎn)和難點(diǎn)。有研究表明,手術(shù)、麻醉引起的中樞神經(jīng)炎癥反應(yīng)是術(shù)后認(rèn)知功能障礙發(fā)生的主要機(jī)制之一[3]。而干擾中樞神經(jīng)系統(tǒng)炎癥反應(yīng)可以改善術(shù)后認(rèn)知功能,降低POCD的發(fā)生率[4-7]。膠質(zhì)細(xì)胞是中樞炎癥反應(yīng)的重要組成部分,其活化后產(chǎn)生的炎癥因子及形態(tài)的改變可誘發(fā)炎癥反應(yīng),直接或間接損傷神經(jīng)元產(chǎn)生神經(jīng)毒性進(jìn)而影響認(rèn)知功能[3,8],如過(guò)度增生的星形膠質(zhì)細(xì)胞可以抑制軸突再生、損傷神經(jīng)組織,過(guò)度活化的小膠質(zhì)細(xì)胞具有神經(jīng)毒性,可以產(chǎn)生并釋放潛在的神經(jīng)毒性物質(zhì)破壞鄰近的神經(jīng)元。有研究表明吸入一定濃度七氟醚可改善認(rèn)知功能[9],也有結(jié)果表明七氟醚對(duì)老年大鼠學(xué)習(xí)記憶相關(guān)的認(rèn)知功能的影響具有劑量依賴性[10-12]。因此,我們預(yù)測(cè)七氟醚可能通過(guò)激活神經(jīng)系統(tǒng)膠質(zhì)細(xì)胞,釋放相關(guān)炎癥因子參與中樞神經(jīng)系統(tǒng)炎癥反應(yīng)而對(duì)認(rèn)知功能產(chǎn)生影響。 本研究擬在復(fù)合脛骨骨折內(nèi)固定術(shù)的基礎(chǔ)上探討誘發(fā)老年大鼠術(shù)后認(rèn)知功能障礙的七氟醚麻醉濃度,并進(jìn)一步探討膠質(zhì)細(xì)胞活化在七氟醚誘發(fā)POCD中的作用機(jī)制,以探究適宜的麻醉深度,為降低臨床中術(shù)后認(rèn)知功能障礙的發(fā)生率提供更多的理論依據(jù)。 第一部分：不同濃度七氟醚麻醉對(duì)老年大鼠術(shù)后認(rèn)知功能的影響 目的：篩選誘發(fā)老年大鼠術(shù)后認(rèn)知功能障礙的七氟醚麻醉濃度。 方法：健康雄性Wistar大鼠96只,體重600～650g,19～20月齡,采用隨機(jī)數(shù)字表法分為6組(n=16)：對(duì)照組(C組)、丙泊酚組(P組)、丙泊酚+手術(shù)組(PS組)、2.4%七氟醚+手術(shù)組(S1組)、3.1%七氟醚+手術(shù)組(S2組)、3.6%七氟醚+手術(shù)組(S3組)。C組、P組、PS組給予吸入30%O22h,S1組、S2組、S3組分別給予吸入2.4%、3.1%、3.6%七氟醚2h,同時(shí)P組、PS組給予丙泊酚(0.5～0.7mg-kg-1·min-1)尾靜脈持續(xù)輸注2h,其余各組以相同速度給予0.9%NaCl注射液尾靜脈持續(xù)輸注2h,PS組、S1組、S2組、S3組在麻醉期間行脛骨骨折切開(kāi)復(fù)位內(nèi)固定手術(shù)。各組分別取8只大鼠于術(shù)后1d、3d、7d分別進(jìn)行Y迷宮空間探索實(shí)驗(yàn)和恐懼條件化實(shí)驗(yàn)測(cè)定大鼠空間工作記憶和恐懼記憶能力,分別于實(shí)驗(yàn)周期開(kāi)始、結(jié)束時(shí)測(cè)定記錄大鼠基礎(chǔ)體重、7日后體重,在整個(gè)實(shí)驗(yàn)周期中,定量觀察大鼠攝食情況,綜合評(píng)估老年大鼠術(shù)后認(rèn)知功能,篩選出誘發(fā)老年大鼠認(rèn)知功能障礙的七氟醚濃度組即SS組。 結(jié)果：與C組比較,其余各組在訓(xùn)練期各個(gè)訓(xùn)練階段的僵直時(shí)間百分比組間差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),PS組、S1組、S2組、S3組表現(xiàn)為飲食量下降,體重下降,(P0.05),其中S3組體重下降明顯,(P0.01),于術(shù)后1d、3d、7d在各臂中的穿梭次數(shù)減少、在新異臂的停留時(shí)間縮短、場(chǎng)景相關(guān)僵直百分比降低、條件誘導(dǎo)僵直百分比降低(P0.05)；與PS組比較,S3組于術(shù)后1d、3d時(shí)穿梭次數(shù)減少及在新異臂停留時(shí)間縮短,于術(shù)后1d時(shí)場(chǎng)景相關(guān)僵直百分比降低,于1d、3d、7d時(shí)條件誘導(dǎo)僵直百分比降低(P0.05)； 結(jié)論：給予高濃度(3.6%,1.5MAC)七氟醚吸入麻醉可誘發(fā)老年大鼠術(shù)后認(rèn)知功能障礙的發(fā)生。 第二部分：探討膠質(zhì)細(xì)胞活化在七氟醚誘發(fā)老年大鼠術(shù)后認(rèn)知功能障礙中相關(guān)調(diào)控機(jī)制 目的：探討膠質(zhì)細(xì)胞活化在七氟醚誘發(fā)老年大鼠術(shù)后認(rèn)知功能中相關(guān)調(diào)控機(jī)制。 方法：健康雄性Wistar大鼠144只,體重600～650g,19～20月齡,隨機(jī)分為4組(n=36)：對(duì)照組(C組)、丙泊酚組(P組)、丙泊酚+手術(shù)組(PS組)、3.6%七氟醚+手術(shù)組(SS組)。于術(shù)后1d、3d、7d分別取6只大鼠,采用免疫熒光法檢測(cè)海馬各亞區(qū)小膠質(zhì)細(xì)胞和星形膠質(zhì)細(xì)胞的形態(tài)及CD68和GFAP的表達(dá)水平,采用Western blot法檢測(cè)海馬IL-1β、IL-6和TNF-α的蛋白表達(dá)水平。 結(jié)果：與C組比較,PS、SS組于術(shù)后1d、3d、7d時(shí)小膠質(zhì)細(xì)胞CD68和星形膠質(zhì)細(xì)胞GFAP、海馬IL-1β、IL-6和TNF-α蛋白表達(dá)增加(P0.05),與PS組比較,SS組在1d、3d時(shí)海馬CD68、GFAP、IL-11β、IL-6和TNF-α蛋白表達(dá)增加(P0.05)。PS、SS組在不同時(shí)點(diǎn)小膠質(zhì)細(xì)胞和星形膠質(zhì)細(xì)胞的形態(tài)發(fā)生變化。 結(jié)論：七氟醚誘發(fā)老年大鼠術(shù)后認(rèn)知功能障礙這一過(guò)程可能與可增強(qiáng)術(shù)后海馬膠質(zhì)細(xì)胞的活化有關(guān)。
[Abstract]:Cognitive dysfunction is a common neurological complication after anesthesia, which has become a major problem affecting the process of recovery and quality of life in the elderly. [1]. sevoflurane is a commonly used inhalation anesthetic, which affects the postoperative cognitive function of [2], but the mechanism is unclear. Therefore, a thorough understanding of the cognitive work of sevoflurane induced operation has been made. It is a hot and difficult point in the research of Anesthesiology that the mechanism of ability to be able to be impaired and to find a suitable depth of sevoflurane anesthesia and to reduce the occurrence of cognitive dysfunction is a hot and difficult point. Response can improve postoperative cognitive function and reduce the incidence of POCD. [4-7]. glial cells are important components of central inflammatory reaction. The inflammatory factors and morphological changes produced after activation can induce inflammatory reactions, and directly or indirectly damage neurons to produce neurotoxicity and then affect cognitive function [3,8], such as hyperproliferative astrocytes. The stromal cells can inhibit axon regeneration, damage the nerve tissue, and the over activated microglia have neurotoxicity, which can produce and release potential neurotoxic substances to destroy adjacent neurons. Some studies have shown that inhaling a certain concentration of sevoflurane can improve cognitive function [9], and the results show that sevoflurane has a learning and memory phase in old rats. The effects of cognitive function are dose dependent [10-12]., so we predict that sevoflurane may affect the cognitive function by activating the nervous system glial cells and releasing related inflammatory factors to participate in the central nervous system inflammatory response.
On the basis of internal fixation of composite tibial fracture, the aim of this study is to explore the anesthetic concentration of sevoflurane induced by postoperative cognitive impairment in old rats, and to further explore the mechanism of the action of glial activation in sevoflurane induced POCD in order to explore the appropriate depth of anesthesia and to reduce the incidence of postoperative cognitive dysfunction. More theoretical basis.
Part I: effects of different concentrations of sevoflurane on postoperative cognitive function in aged rats
Objective: to screen sevoflurane anesthesia for inducing postoperative cognitive dysfunction in aged rats.
Methods: 96 healthy male Wistar rats, weighing 600 to 650g, 19~20 months old, were divided into 6 groups (n=16): control group (group C), propofol group (group P), propofol + operation group (group PS), 2.4% sevoflurane + operation group (group S1), 3.1% sevoflurane + operation group (S2 group), 3.6% sevoflurane + operation group (S3 group).C group, P group, PS group was given inhalation 22h, group S1, group S2, and group S3 were given 2.4%, 3.1%, 3.6% sevoflurane 2h, and group P, PS group was given propofol (0.5 ~ 0.7mg-kg-1 min-1) tail vein for continuous infusion of 2H. In each group, 8 rats were taken at 1D, 3D, and 7d after the operation. The spatial working memory and fear memory ability of the rats were measured by the Y maze experiment and the fear conditioned experiment respectively. At the beginning of the experiment, the basal body weight of the rats was recorded at the end of the experiment. The weight of the rats was recorded after 7 days, and the feeding situation of the rats was observed in the whole experimental period. Combined assessment of postoperative cognitive function in elderly rats, sevoflurane concentration group induced by cognitive impairment in elderly rats was screened out, namely SS group.
Results: compared with the C group, there was no significant difference between the other groups at each training stage in the training phase (P0.05). Group PS, group S1, S2 group, and S3 group showed decreased diet and weight loss, (P0.05), and S3 group weight decreased obviously, (P0.01), 1D, 3D, and 7d in the different arms in the new arm The retention time was shortened, the percentage of the scene related stiffness decreased and the percentage of conditional stiffness decreased (P0.05). Compared with the PS group, the number of shuttles in the S3 group decreased at 1D, 3D and the length of the new arm was shortened, and the relative stiffness percentage decreased at 1D after operation, and the percentage of stiffness induced stiffness decreased (P0.05) at 1D, 3D and 7d.
Conclusion: high concentration (3.6%, 1.5MAC) sevoflurane inhalation anesthesia can induce postoperative cognitive dysfunction in aged rats.
The second part: To explore the mechanism of glial cell activation in sevoflurane induced postoperative cognitive dysfunction in aged rats.
Objective: To explore the mechanism of glial cell activation in sevoflurane induced cognitive function in aged rats.
Methods: 144 healthy male Wistar rats, weighing 600 to 650g and 19~20 months old, were randomly divided into 4 groups (n=36): control group (group C), propofol group (group P), propofol + operation group (group PS), 3.6% sevoflurane + operation group (group SS). 6 rats were taken 1D, 3D and 7d respectively after operation. The microglia and astrocytes in hippocampus subregions were detected by immunofluorescence. The expression of CD68 and GFAP in hippocampus was detected by Western blot.
Results: compared with the C group, PS and SS groups were at 1D, 3D, and 7d in the microglia CD68 and astrocytes GFAP, and the expression of IL-1 beta, IL-6 and TNF- alpha protein in the hippocampus increased (P0.05). The morphology of the cell changes.
CONCLUSION: Sevoflurane-induced cognitive impairment in aged rats may be related to the enhancement of hippocampal glial cell activation.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R614
本文編號(hào)：2161059
[Abstract]:Cognitive dysfunction is a common neurological complication after anesthesia, which has become a major problem affecting the process of recovery and quality of life in the elderly. [1]. sevoflurane is a commonly used inhalation anesthetic, which affects the postoperative cognitive function of [2], but the mechanism is unclear. Therefore, a thorough understanding of the cognitive work of sevoflurane induced operation has been made. It is a hot and difficult point in the research of Anesthesiology that the mechanism of ability to be able to be impaired and to find a suitable depth of sevoflurane anesthesia and to reduce the occurrence of cognitive dysfunction is a hot and difficult point. Response can improve postoperative cognitive function and reduce the incidence of POCD. [4-7]. glial cells are important components of central inflammatory reaction. The inflammatory factors and morphological changes produced after activation can induce inflammatory reactions, and directly or indirectly damage neurons to produce neurotoxicity and then affect cognitive function [3,8], such as hyperproliferative astrocytes. The stromal cells can inhibit axon regeneration, damage the nerve tissue, and the over activated microglia have neurotoxicity, which can produce and release potential neurotoxic substances to destroy adjacent neurons. Some studies have shown that inhaling a certain concentration of sevoflurane can improve cognitive function [9], and the results show that sevoflurane has a learning and memory phase in old rats. The effects of cognitive function are dose dependent [10-12]., so we predict that sevoflurane may affect the cognitive function by activating the nervous system glial cells and releasing related inflammatory factors to participate in the central nervous system inflammatory response.
On the basis of internal fixation of composite tibial fracture, the aim of this study is to explore the anesthetic concentration of sevoflurane induced by postoperative cognitive impairment in old rats, and to further explore the mechanism of the action of glial activation in sevoflurane induced POCD in order to explore the appropriate depth of anesthesia and to reduce the incidence of postoperative cognitive dysfunction. More theoretical basis.
Part I: effects of different concentrations of sevoflurane on postoperative cognitive function in aged rats
Objective: to screen sevoflurane anesthesia for inducing postoperative cognitive dysfunction in aged rats.
Methods: 96 healthy male Wistar rats, weighing 600 to 650g, 19~20 months old, were divided into 6 groups (n=16): control group (group C), propofol group (group P), propofol + operation group (group PS), 2.4% sevoflurane + operation group (group S1), 3.1% sevoflurane + operation group (S2 group), 3.6% sevoflurane + operation group (S3 group).C group, P group, PS group was given inhalation 22h, group S1, group S2, and group S3 were given 2.4%, 3.1%, 3.6% sevoflurane 2h, and group P, PS group was given propofol (0.5 ~ 0.7mg-kg-1 min-1) tail vein for continuous infusion of 2H. In each group, 8 rats were taken at 1D, 3D, and 7d after the operation. The spatial working memory and fear memory ability of the rats were measured by the Y maze experiment and the fear conditioned experiment respectively. At the beginning of the experiment, the basal body weight of the rats was recorded at the end of the experiment. The weight of the rats was recorded after 7 days, and the feeding situation of the rats was observed in the whole experimental period. Combined assessment of postoperative cognitive function in elderly rats, sevoflurane concentration group induced by cognitive impairment in elderly rats was screened out, namely SS group.
Results: compared with the C group, there was no significant difference between the other groups at each training stage in the training phase (P0.05). Group PS, group S1, S2 group, and S3 group showed decreased diet and weight loss, (P0.05), and S3 group weight decreased obviously, (P0.01), 1D, 3D, and 7d in the different arms in the new arm The retention time was shortened, the percentage of the scene related stiffness decreased and the percentage of conditional stiffness decreased (P0.05). Compared with the PS group, the number of shuttles in the S3 group decreased at 1D, 3D and the length of the new arm was shortened, and the relative stiffness percentage decreased at 1D after operation, and the percentage of stiffness induced stiffness decreased (P0.05) at 1D, 3D and 7d.
Conclusion: high concentration (3.6%, 1.5MAC) sevoflurane inhalation anesthesia can induce postoperative cognitive dysfunction in aged rats.
The second part: To explore the mechanism of glial cell activation in sevoflurane induced postoperative cognitive dysfunction in aged rats.
Objective: To explore the mechanism of glial cell activation in sevoflurane induced cognitive function in aged rats.
Methods: 144 healthy male Wistar rats, weighing 600 to 650g and 19~20 months old, were randomly divided into 4 groups (n=36): control group (group C), propofol group (group P), propofol + operation group (group PS), 3.6% sevoflurane + operation group (group SS). 6 rats were taken 1D, 3D and 7d respectively after operation. The microglia and astrocytes in hippocampus subregions were detected by immunofluorescence. The expression of CD68 and GFAP in hippocampus was detected by Western blot.
Results: compared with the C group, PS and SS groups were at 1D, 3D, and 7d in the microglia CD68 and astrocytes GFAP, and the expression of IL-1 beta, IL-6 and TNF- alpha protein in the hippocampus increased (P0.05). The morphology of the cell changes.
CONCLUSION: Sevoflurane-induced cognitive impairment in aged rats may be related to the enhancement of hippocampal glial cell activation.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R614
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