摘要
目的评价氢对大鼠肝脏冷缺血再灌注时内质网应激的影响。方法SPF级健康成年雄性SD大鼠24只，体重200～250 g，采用随机数字表法，将其分为3组（n＝8）：假手术组（S组）、肝脏冷缺血再灌注组（I/R组）和含饱和氢乳酸钠林格液组（HL组）。阻断肝血流后HL组经门静脉灌注4 ℃饱和氢乳酸钠林格液6～8 ml/min，灌注时间为30 min。灌注结束后恢复肝血流。于再灌注6 h（S组于关腹后6 h）时，取血标本，检测血清ALT和AST浓度；取肝组织，行肝组织病理学评分，采用TUNEL法确定细胞凋亡率；采用硫代巴比妥酸法测定MDA含量，采用黄嘌呤氧化酶法测定SOD活性；免疫组化法检测内质网分子伴侣蛋白46（ERP46）、免疫球蛋白重链结合蛋白（BiP）和caspase-12的表达；实时定量PCR法检测肝组织ERP46、BiP和caspase-12的mRNA表达。结果与S组比较，I/R组和HL组血清ALT和AST浓度、肝组织病理学评分、细胞凋亡率和MDA含量升高，SOD活性降低，ERP46、BiP、caspase-12及其mRNA表达上调（P〈0.05）；与I/R组比较，HL组血清ALT和AST浓度、肝组织病理学评分、细胞凋亡率和MDA含量降低，SOD活性升高，ERP46、BiP、caspase-12及其mRNA表达下调（P〈0.05）。结论氢减轻大鼠肝脏冷缺血再灌注时细胞凋亡的机制可能与抑制内质网应激有关。
Objective To evaluate the effect of hydrogen on the endoplasmic reticulum stress during liver cold ischemia-reperfusion（I/R）in rats.Methods Twenty-four SPF healthy adult male Sprague-Dawley rats, weighing 200-250 g, were divided into 3 groups（n=8 each）using a random number table： sham operation group（group S）, liver cold I/R group（group I/R）and hydrogen-saturated lactated Ringer′s solution group（group HL）. After occlusion of the liver blood flow, livers were perfused through the portal vein with 4 ℃ hydrogen-saturated lactated Ringer′s solution 6-8 ml/min for 30 min.The liver blood flow was restored after the end of perfusion.At 6 h of reperfusion in I/R and HL groups or at 6 h after peritoneum closure in group S, blood samples from the inferior vena cava were collected for determination of serum alanine transaminase（ALT）and aspartate transaminase（AST）concentrations.After blood sampling, liver tissues were obtained for examination of pathological changes and for determination of cell apoptosis（using TUNEL）, malondialdehyde（MDA）content（by thiobarbituric acid method）, superoxide dismutase（SOD）activity（using xanthine oxidase method）, expression of endoplasmic reticulum protein 46（ERP46）, immunoglobulin heavy chain binding protein（BiP）and caspase-12（by immunohistochemistry）, and expression of ERP46, BiP and caspase-12 mRNA（using quantitative real-time polymerase chain reaction）. The pathological changes were scored.Apoptosis rate was calculated.Results Compared with group S, the serum ALT and AST concentrations, pathological scores, apoptosis rate and MDA content were significantly increased, the SOD activity was decreased, and the expression of ERP46, BiP and caspase-12 protein and mRNA was up-regulated in I/R and HL groups（P〈0.05）. Compared with group I/R, the serum ALT and AST concentrations, pathological scores, apoptosis rate and MDA content were significantly decreased, the SOD activity was increased, and the expression of ERP46, BiP
出处
《中华麻醉学杂志》
CSCD
北大核心
2017年第4期450-453,共4页
Chinese Journal of Anesthesiology
关键词
氢
内质网
应激
肝移植
再灌注损伤
Hydrogen
Endoplasmic recticulum
Stress
Liver transplantation
Reperfusion injury
