七氟醚后处理对犬体外循环肺损伤及AktNF-κB表达的影响

发布时间：2018-01-08 22:16

本文关键词：七氟醚后处理对犬体外循环肺损伤及AktNF-κB表达的影响　出处：《遵义医学院》2015年硕士论文　论文类型：学位论文

【摘要】：目的:研究七氟醚后处理对体外循环(CPB)犬肺组织蛋白质丝氨酸苏氨酸激酶(Akt)/核转录因子-κB(NF-κB)表达的影响,探讨七氟醚后处理减轻CPB肺损伤的作用机制。方法:健康成年杂种犬24只随机分为四组,每组6只(n=6):单纯手术组(C组)、体外循环缺血再灌注组(R组)、七氟醚后处理组(S组)、渥曼青霉素(wortmannin)+七氟醚后处理组(W组)。2.5%戊巴比妥钠腹腔内注射麻醉固定后,行双腔气管插管机械通气,右侧股动、静脉置管监测有创动脉血压及中心静脉压。正中开胸,全身肝素化后分别行左锁骨下动脉和右心耳插管,连接人工心肺机及CPB管道,分离左肺动脉,建立犬体外循环单肺缺血再灌注损伤模型。S组在开放左肺动脉即刻通过人工膜肺给予呼气末2%七氟醚后处理30min,W组在开放左肺动脉前10min给予wortmannin,开放左肺动脉的同时,给予2%七氟醚后处理30min,C组只开胸,不做其他处理。各组犬于CPB前(T1)、开放左肺动脉后(T2)及停机后2h(T3)三个时间点抽取股动脉血行血气分析计算氧合指数(OI)和呼吸指数(RI);取左肺组织保存,HE染色光镜观察形态学变化;石蜡固定后TUNEL法检测细胞凋亡情况;采用ELISA法检测肺组织TNF-α的含量;应用Western blot技术检测肺组织Akt、p-Akt(Ser473)及NF-κB(P65)蛋白表达变化。结果:(1)肺功能指标的变化:四组内不同时点比较:C组各时点OI、RI未见明显改变(P0.05);与T1比较,T2、T3时刻R、S、W三组OI显著降低,而RI则明显升高(P0.05);与T2时刻比较,T3时刻R、S、W三组OI明显降低,RI明显升高(P0.05)。四组间同时点比较:T1时刻:四组犬之间OI、RI无明显差异(P0.05);T2时刻:与C组比较,R、S、W三组OI显著减低,RI则明显升高(P0.05);T3时刻,R、S、W三组OI低于C组,相反RI明显升高(P0.05),S组OI明显高于R和W组,而RI则相反(P0.05),R和W组之间OI、RI无明显差异(P0.05)。(2)肺组织光镜形态学观察:四组犬肺组织损伤程度随时间延长逐渐加重。各组T1时刻肺组织结构完整清晰;T2时刻四组犬肺组织结构稍紊乱,可见部分肺泡壁断裂,出现炎性细胞浸润;T3时刻C组肺组织结构较清晰,见少量肺泡壁断裂及炎性渗出;R和W组肺组织结构紊乱,肺泡壁断裂严重,肺泡腔塌陷充满水肿液,可见大量炎性细胞和红细胞浸润;S组肺组织结构尚清晰,部分肺泡腔塌陷,肺泡壁断裂,可见炎性细胞及红细胞渗出,肺损伤程度明显较R和W组轻。(3)测肺组织细胞凋亡及IHS评分:四组犬肺组织凋亡程度随时间延长逐渐增加。T1时刻各组犬肺组织可见少量棕褐色的凋亡细胞;T2时刻凋亡细胞数目逐渐增多,染色也逐渐加深;T3时刻,C组出现多量染色较深的阳性细胞,R、S、W三组均出现大量的深染阳性细胞,R、W两组尤甚并形成凋亡细胞群,凋亡程度明显高于S组。免疫组化评分(IHS):四组内各时点比较:C组各时点肺组织IHS评分无明显变化;与T1比较,T3时刻R、S、W三组IHS评分均升高(P0.05),;与T2比较,T3时刻R、S、W三组IHS评分明显升高(P0.05)。四组间同时点比较:T1、T2时刻各组犬之间IHS评分无明显差异(P0.05);T3时刻,R、S、W三组IHS评分明显高于C组(P0.05),与S组比较,R组和W组IHS评分明显升高(P0.05),R组和W组IHS评分无明显差异(P0.05)。(4)肺组织Akt/p-Akt(Ser473)表达变化:肺组织总Akt表达变化:四组内各时点及四组之间同时点比较,犬肺组织总Akt表达无明显差异(P0.05)。肺组织p-Akt(Ser473)表达变化:四组内各时点比较:C组各时点肺p-Akt(Ser473)表达未见明显改变(P0.05);与T1比较,T2时刻R、S、W三组肺组织p-Akt表达均升高(P0.05),R组和S组T3时刻p-Akt表达明显高于T1时刻(P0.05),W组表达变化不显著(P0.05);与T2比较,T3时刻R、S两组肺组织p-Akt表达水平增高(P0.05),W组肺组织p-Akt表达水平降低(P0.05)。四组间同时点比较:T1时刻四组犬肺组织p-Akt表达无明显差异(P0.05);T2时刻:与C组比较,R、S、W三组肺组织p-Akt表达增加(P0.05);T3时刻:S组和R组肺组织p-Akt表达水平明显高于C组(P0.05),W组与C组比较无统计学意义(P0.05),S组p-Akt表达量显著高于R组(P0.05)。(5)肺组织NF-κB(P65)表达的变化:四组内各时点比较:C组各时间点未见明显改变;与T1比较,T2、T3时刻R、S、W三组肺组织NF-κB表达水平明显升高(P0.05);与T2时刻比较,T3时刻R、S、W三组犬肺组织NF-κB表达水平均升高(P0.05)。四组间同时点比较:T1时刻,四组犬肺组织NF-κB表达无明显差异(P0.05);T2时刻:与C组比较,R、S、W三组犬肺组织NF-κB水平升高(P0.05);T3时刻:C组犬肺组织NF-κB表达水平明显低于R、S、W三组(P0.05),S组肺组织NF-κB表达水平均明显低于R和W组(P0.05),R组和W组NF-κB表达无明显差异(P0.05)。(6)肺组织TNF-α含量的变化:四组内各时点比较:C组各时点肺组织TNF-α含量未见明显改变(P0.05);与T1比较,T2、T3时刻R、S、W三组肺组织TNF-α含量明显增加(P0.05);与T2时刻比较,T3时刻R、S、W三组犬肺组织TNF-α含量明显升高(P0.05)。四组间同时点比较:T1时刻四组犬肺组织TNF-α含量无明显差异(P0.05);T2时刻:与C组比较,R、S、W三组肺组织TNF-α含量明显增加(P0.05);T3时刻:R、S、W三组肺组织TNF-α含量明显高于C组(P0.05),S组TNF-α含量明显低于R和W组(P0.05),R组和W组TNF-α含量无明显差异(P0.05)。结论:(1)CPB后随着犬肺组织TNF-α含量的增加,细胞凋亡增加,肺组织结构破坏严重,肺功能降低,肺组织TNF-α含量的增加可能是导致细胞凋亡,加重肺损伤的重要原因。(2)七氟醚后处理后肺功能明显改善,细胞凋亡减少、肺组织破坏程度减轻,对CPB犬肺损伤有一定的保护作用。(3)七氟醚后处理减轻CPB肺损伤的作用可能与激活PI3K/Akt信号通路抑制肺组织TNF-α和NF-κB表达有关。
[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 - 伪 in lung tissues of three groups were significantly higher than those in group C ( P0.05 ) , and the levels of TNF - 伪 in lung tissues were significantly higher than those in group C ( P0.05 ) .

【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R614

【共引文献】