發(fā)布時(shí)間：2018-01-08 22:16

  本文關(guān)鍵詞：七氟醚后處理對(duì)犬體外循環(huán)肺損傷及AktNF-κB表達(dá)的影響　出處：《遵義醫(yī)學(xué)院》2015年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 七氟醚后處理 體外循環(huán) NF-κB Akt TNF-α

【摘要】：目的:研究七氟醚后處理對(duì)體外循環(huán)(CPB)犬肺組織蛋白質(zhì)絲氨酸蘇氨酸激酶(Akt)/核轉(zhuǎn)錄因子-κB(NF-κB)表達(dá)的影響,探討七氟醚后處理減輕CPB肺損傷的作用機(jī)制。方法:健康成年雜種犬24只隨機(jī)分為四組,每組6只(n=6):單純手術(shù)組(C組)、體外循環(huán)缺血再灌注組(R組)、七氟醚后處理組(S組)、渥曼青霉素(wortmannin)+七氟醚后處理組(W組)。2.5%戊巴比妥鈉腹腔內(nèi)注射麻醉固定后,行雙腔氣管插管機(jī)械通氣,右側(cè)股動(dòng)、靜脈置管監(jiān)測(cè)有創(chuàng)動(dòng)脈血壓及中心靜脈壓。正中開(kāi)胸,全身肝素化后分別行左鎖骨下動(dòng)脈和右心耳插管,連接人工心肺機(jī)及CPB管道,分離左肺動(dòng)脈,建立犬體外循環(huán)單肺缺血再灌注損傷模型。S組在開(kāi)放左肺動(dòng)脈即刻通過(guò)人工膜肺給予呼氣末2%七氟醚后處理30min,W組在開(kāi)放左肺動(dòng)脈前10min給予wortmannin,開(kāi)放左肺動(dòng)脈的同時(shí),給予2%七氟醚后處理30min,C組只開(kāi)胸,不做其他處理。各組犬于CPB前(T1)、開(kāi)放左肺動(dòng)脈后(T2)及停機(jī)后2h(T3)三個(gè)時(shí)間點(diǎn)抽取股動(dòng)脈血行血?dú)夥治鲇?jì)算氧合指數(shù)(OI)和呼吸指數(shù)(RI);取左肺組織保存,HE染色光鏡觀察形態(tài)學(xué)變化;石蠟固定后TUNEL法檢測(cè)細(xì)胞凋亡情況;采用ELISA法檢測(cè)肺組織TNF-α的含量;應(yīng)用Western blot技術(shù)檢測(cè)肺組織Akt、p-Akt(Ser473)及NF-κB(P65)蛋白表達(dá)變化。結(jié)果:(1)肺功能指標(biāo)的變化:四組內(nèi)不同時(shí)點(diǎn)比較:C組各時(shí)點(diǎn)OI、RI未見(jiàn)明顯改變(P0.05);與T1比較,T2、T3時(shí)刻R、S、W三組OI顯著降低,而RI則明顯升高(P0.05);與T2時(shí)刻比較,T3時(shí)刻R、S、W三組OI明顯降低,RI明顯升高(P0.05)。四組間同時(shí)點(diǎn)比較:T1時(shí)刻:四組犬之間OI、RI無(wú)明顯差異(P0.05);T2時(shí)刻:與C組比較,R、S、W三組OI顯著減低,RI則明顯升高(P0.05);T3時(shí)刻,R、S、W三組OI低于C組,相反RI明顯升高(P0.05),S組OI明顯高于R和W組,而RI則相反(P0.05),R和W組之間OI、RI無(wú)明顯差異(P0.05)。(2)肺組織光鏡形態(tài)學(xué)觀察:四組犬肺組織損傷程度隨時(shí)間延長(zhǎng)逐漸加重。各組T1時(shí)刻肺組織結(jié)構(gòu)完整清晰;T2時(shí)刻四組犬肺組織結(jié)構(gòu)稍紊亂,可見(jiàn)部分肺泡壁斷裂,出現(xiàn)炎性細(xì)胞浸潤(rùn);T3時(shí)刻C組肺組織結(jié)構(gòu)較清晰,見(jiàn)少量肺泡壁斷裂及炎性滲出;R和W組肺組織結(jié)構(gòu)紊亂,肺泡壁斷裂嚴(yán)重,肺泡腔塌陷充滿(mǎn)水腫液,可見(jiàn)大量炎性細(xì)胞和紅細(xì)胞浸潤(rùn);S組肺組織結(jié)構(gòu)尚清晰,部分肺泡腔塌陷,肺泡壁斷裂,可見(jiàn)炎性細(xì)胞及紅細(xì)胞滲出,肺損傷程度明顯較R和W組輕。(3)測(cè)肺組織細(xì)胞凋亡及IHS評(píng)分:四組犬肺組織凋亡程度隨時(shí)間延長(zhǎng)逐漸增加。T1時(shí)刻各組犬肺組織可見(jiàn)少量棕褐色的凋亡細(xì)胞;T2時(shí)刻凋亡細(xì)胞數(shù)目逐漸增多,染色也逐漸加深;T3時(shí)刻,C組出現(xiàn)多量染色較深的陽(yáng)性細(xì)胞,R、S、W三組均出現(xiàn)大量的深染陽(yáng)性細(xì)胞,R、W兩組尤甚并形成凋亡細(xì)胞群,凋亡程度明顯高于S組。免疫組化評(píng)分(IHS):四組內(nèi)各時(shí)點(diǎn)比較:C組各時(shí)點(diǎn)肺組織IHS評(píng)分無(wú)明顯變化;與T1比較,T3時(shí)刻R、S、W三組IHS評(píng)分均升高(P0.05),;與T2比較,T3時(shí)刻R、S、W三組IHS評(píng)分明顯升高(P0.05)。四組間同時(shí)點(diǎn)比較:T1、T2時(shí)刻各組犬之間IHS評(píng)分無(wú)明顯差異(P0.05);T3時(shí)刻,R、S、W三組IHS評(píng)分明顯高于C組(P0.05),與S組比較,R組和W組IHS評(píng)分明顯升高(P0.05),R組和W組IHS評(píng)分無(wú)明顯差異(P0.05)。(4)肺組織Akt/p-Akt(Ser473)表達(dá)變化:肺組織總Akt表達(dá)變化:四組內(nèi)各時(shí)點(diǎn)及四組之間同時(shí)點(diǎn)比較,犬肺組織總Akt表達(dá)無(wú)明顯差異(P0.05)。肺組織p-Akt(Ser473)表達(dá)變化:四組內(nèi)各時(shí)點(diǎn)比較:C組各時(shí)點(diǎn)肺p-Akt(Ser473)表達(dá)未見(jiàn)明顯改變(P0.05);與T1比較,T2時(shí)刻R、S、W三組肺組織p-Akt表達(dá)均升高(P0.05),R組和S組T3時(shí)刻p-Akt表達(dá)明顯高于T1時(shí)刻(P0.05),W組表達(dá)變化不顯著(P0.05);與T2比較,T3時(shí)刻R、S兩組肺組織p-Akt表達(dá)水平增高(P0.05),W組肺組織p-Akt表達(dá)水平降低(P0.05)。四組間同時(shí)點(diǎn)比較:T1時(shí)刻四組犬肺組織p-Akt表達(dá)無(wú)明顯差異(P0.05);T2時(shí)刻:與C組比較,R、S、W三組肺組織p-Akt表達(dá)增加(P0.05);T3時(shí)刻:S組和R組肺組織p-Akt表達(dá)水平明顯高于C組(P0.05),W組與C組比較無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),S組p-Akt表達(dá)量顯著高于R組(P0.05)。(5)肺組織NF-κB(P65)表達(dá)的變化:四組內(nèi)各時(shí)點(diǎn)比較:C組各時(shí)間點(diǎn)未見(jiàn)明顯改變;與T1比較,T2、T3時(shí)刻R、S、W三組肺組織NF-κB表達(dá)水平明顯升高(P0.05);與T2時(shí)刻比較,T3時(shí)刻R、S、W三組犬肺組織NF-κB表達(dá)水平均升高(P0.05)。四組間同時(shí)點(diǎn)比較:T1時(shí)刻,四組犬肺組織NF-κB表達(dá)無(wú)明顯差異(P0.05);T2時(shí)刻:與C組比較,R、S、W三組犬肺組織NF-κB水平升高(P0.05);T3時(shí)刻:C組犬肺組織NF-κB表達(dá)水平明顯低于R、S、W三組(P0.05),S組肺組織NF-κB表達(dá)水平均明顯低于R和W組(P0.05),R組和W組NF-κB表達(dá)無(wú)明顯差異(P0.05)。(6)肺組織TNF-α含量的變化:四組內(nèi)各時(shí)點(diǎn)比較:C組各時(shí)點(diǎn)肺組織TNF-α含量未見(jiàn)明顯改變(P0.05);與T1比較,T2、T3時(shí)刻R、S、W三組肺組織TNF-α含量明顯增加(P0.05);與T2時(shí)刻比較,T3時(shí)刻R、S、W三組犬肺組織TNF-α含量明顯升高(P0.05)。四組間同時(shí)點(diǎn)比較:T1時(shí)刻四組犬肺組織TNF-α含量無(wú)明顯差異(P0.05);T2時(shí)刻:與C組比較,R、S、W三組肺組織TNF-α含量明顯增加(P0.05);T3時(shí)刻:R、S、W三組肺組織TNF-α含量明顯高于C組(P0.05),S組TNF-α含量明顯低于R和W組(P0.05),R組和W組TNF-α含量無(wú)明顯差異(P0.05)。結(jié)論:(1)CPB后隨著犬肺組織TNF-α含量的增加,細(xì)胞凋亡增加,肺組織結(jié)構(gòu)破壞嚴(yán)重,肺功能降低,肺組織TNF-α含量的增加可能是導(dǎo)致細(xì)胞凋亡,加重肺損傷的重要原因。(2)七氟醚后處理后肺功能明顯改善,細(xì)胞凋亡減少、肺組織破壞程度減輕,對(duì)CPB犬肺損傷有一定的保護(hù)作用。(3)七氟醚后處理減輕CPB肺損傷的作用可能與激活PI3K/Akt信號(hào)通路抑制肺組織TNF-α和NF-κB表達(dá)有關(guān)。
[Abstract]:Objective : To study the effect of post - treatment of heptafluoroether on the expression of protein serine - threonine kinase ( PTK ) / nuclear transcription factor - 魏B ( NF - 魏B ) in canine lung tissue during cardiopulmonary bypass ( CPB ) . There was no significant difference in OI and RI between four groups ( P0.05 ) . The levels of OI and RI in groups R , S and W were significantly lower than those in group C ( P0.05 ) . At the same time , the number of apoptotic cells in group C was significantly higher than that in group C ( P0.05 ) . At the same time , the levels of R , S and W in group C were significantly higher than those in group C ( P0.05 ) . There was no significant difference in the expression of p - protein in lung tissue ( P0.05 ) . At the same time , there was no significant difference in the expression level of p - protein in lung tissue ( P0.05 ) in group C and group C ( P0.05 ) . ( 5 ) The expression of NF - 魏B ( P65 ) in lung tissue was significantly higher than that in group C ( P0.05 ) . The levels of NF - 魏B in lung tissues of three groups were significantly lower than those in group C ( P0.05 ) . Compared with group C , the levels of TNF - 偽 in lung tissues of three groups were significantly higher than those in group C ( P0.05 ) , and the levels of TNF - 偽 in lung tissues were significantly higher than those in group C ( P0.05 ) .

【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：R614

【共引文獻(xiàn)】

相關(guān)博士學(xué)位論文 前10條

1 王麗s，

本文編號(hào)：1398889