小檗碱对大鼠脂肪肝缺血再灌注损伤的影响:与肝组织内质网应激的关系  

Effect of berberine on fatty liver ischemia-reperfusion injury in rats:the relationship with endoplasmic reticulum stress in liver tissues

作　　者：张楠[1] 盛明薇[2] 张馨月 乌曼 丁艺洁 喻文立[2] 杜洪印[2] Zhang Nan;Sheng Mingwei;Zhang Xinyue;Wu Man;Ding Yijie;Yu Wenli;Du Hongyin(First Central Clinical College of Tianjin Medical University,Tianjin 300192,China;Department of Anesthesiology,Tianjin First Central Hospital,Tianjin 300192,China)

机构地区：[1]天津医科大学一中心临床学院,300192 [2]天津市第一中心医院麻醉科,300192

出　　处：《中华麻醉学杂志》2019年第2期162-166,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81700659);天津市自然科学基金(17JCYBJC28000);天津市卫生行业重点攻关项目(16KG101,13KG105);天津市卫生和计划生育委员会中医中西医结合科研课题(2017056).

摘　　要：目的评价小檗碱对大鼠脂肪肝缺血再灌注损伤的影响及其与肝组织内质网应激的关系。方法雄性Wistar大鼠32只,4周龄,体重为100~150g,采用随机数字表法分为4组(n=8):对照组(C组)正常饮食12周,第9周开始生理盐水3.5 ml灌胃4周,仅行开关腹处理;脂肪肝组(FL组)高脂饮食(45%脂肪热能)12周,余同C组处理;缺血再灌注组(I/R组)高脂饮食(45%脂肪热能)12周,第9周开始生理盐水3.5ml灌胃4周,随后建立大鼠肝缺血再灌注损伤模型;小檗碱组(BBR组)高脂饮食(45%脂肪热能)12周,第9周开始小檗碱溶液(300 mg/kg)3.5 ml灌胃4周,随后建立大鼠肝缺血再灌注损伤模型。采用阻断门静脉、肝固有动脉、胃右静脉、肝上下腔静脉和肝下下腔静脉进行4℃乳酸钠林格液冷灌注30 min后再灌注的方法制备肝缺血再灌注损伤模型。于喂养4、8、12周时取血,测定血清甘油三酯(TG)浓度。再灌注6 h时取血,测定血清AST和ALT浓度。再灌注6 h取血结束后,取肝组织,行油红O染色和HE染色。采用Western blot法测定肝组织内质网应激蛋白免疫球蛋白重链结合蛋白质(Bip)、CCAAT/增强子结合蛋白(CHOP)、蛋白激酶RNA样内质网激酶(PERK)和磷酸化PERK(p-PERK)的表达水平。计算p-PERK/PERK比值。结果与C组相比,FL组血清TG、ALT和AST浓度升高,肝组织Bip和CHOP表达上调,p-PERK/PERK比值升高(P<0.05),脂质沉积增多,呈现脂肪变性。与FL组相比,I/R组血清ALT和AST浓度升高,肝组织Bip和CHOP表达上调,p-PERK/PERK比值升高(P<0.05),肝组织病理学损伤加重。与I/R组相比,BBR组血清TG、ALT和AST浓度降低,肝组织Bip和CHOP表达下调,p-PERK/PERK比值降低(P<0.05),脂质沉积减低,肝组织病理学损伤减轻。结论小檗碱可减轻大鼠脂肪肝缺血再灌注损伤,机制可能与抑制内质网应激有关。Objective To evaluate the effect of berberine on fatty liver ischemia-reperfusion(I/R)injury and the relationship with endoplasmic reticulum stress in liver tissues.Methods Thirty-two male Wistar rats,aged 4 weeks,weighing 100-150 g,were divided into 4 groups(n=8 each)using a random number table method:control group(group C),fatty liver group(group FL),I/R group and berberine group(group BBR).Rats were fed a normal fat diet for 12 weeks,normal saline 3.5 ml was given intragastrically for 4 weeks starting from 9th week,and rats only underwent simple laparotomy in group C.Rats were fed a high-fat diet(45%energy originating from fat)for 12 weeks,and the other treatments were similar to those previously described in group C.Rats were fed a high-fat diet(45%energy originating from fat)for 12 weeks,normal saline 3.5 ml was given intragastrically for 4 weeks starting from 9th week,and then the model of liver I/R injury was established in group I/R.Rats were fed a high-fat diet(45%energy originating from fat)for 12 weeks,berberine solution(300 mg/kg)3.5 ml was given intragastrically for 4 weeks starting from 9th week,and then the model of liver I/R injury was established in group BBR.Hepatic ischemia was induced by clamping the portal vein,hepatic artery,right gastric vein,and supra-and inferior-hepatic vena cava to perform cold perfusion with 4℃lactated Ringer′s solution lasting for 30 min,followed by reperfusion.The serum triglyceride(TG)concentrations were determined after 4,8 and 12 weeks of diet.Blood samples were collected at 6 h of reperfusion for measurement of serum aspartate transminase(AST)and alanine transaminase(ALT)concentrations.Livers were removed after blood sampling at 6 h of reperfusion and liver tissues were obtained and stained with oil red O and haematoxylin and eosin for examination of pathological changes and for determination of the expression of Bip,CCAAT/enhancer-binding protein homologous protein(CHOP),protein kinase RNA-like endoplasmic reticulum kinase(PERK)and phosphorylated PERK(p-PERK

关 键 词：小檗碱 脂肪肝 再灌注损伤 内质网 应激 

分 类 号：R657.3[医药卫生—外科学]