沈艳艳,任婷婷
SHEN Yanyan,REN Tingting

• 1:贵州医科大学医学检验学院临床生物化学教研室

摘要(Abstract)：

目的:分析蓝莓及蓝莓联合益生菌对酒精性脂肪性肝病(AFLD)肝损伤的保护作用及机制。方法:58只C57BL/6J小鼠分为正常控制组(CG组)、模型组(MG组)、蓝莓干预组(BJ组)、蓝莓联合益生菌干预组(BJB组)、SIRT1 siRNA阻断组(SI组)、蓝莓+SIRT1阻断组(BJSI组)及蓝莓联合益生菌+SIRT1阻断组(BJBSI组),MG组为10只、其余每组8只;除CG组外,其余6组喂饲利伯De Carli'82饲料复制AFLD模型(所有小鼠均于造模成功后改喂普通饲料),喂饲造模饲料前7 d BJ组、BJB组小鼠分别进行蓝莓、蓝莓联合益生菌预防灌胃;SI组造模成功后,给予慢病毒质粒包装SIRT1 siRNA腹部肝区注射; BJSI组、BJBSI组小鼠造模成功后,分别进行蓝莓、蓝莓联合益生菌干预,同时慢病毒质粒包装SIRT1 siRNA腹部肝区注射;造模喂饲12周时,采用病理学方法确认造模情况,造模成功后10 d时各组小鼠眼框取血、颈椎脱臼处死后取肝组织,采用病理学方法观察各组小鼠肝组织学改变,生化方法检测血清甘油三酯(TG)、总胆固醇(TC)、丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、高密度脂蛋白胆固醇(HDL-c)及低密度脂蛋白胆固醇(LDL-c)水平,ELISA法检测小鼠肝组织丙二醛(MDA)、还原型谷胱甘肽(GSH)、超氧化物歧化酶(SOD)水平及沉默信息调节因子1(SIRT1)蛋白表达水平。结果:病理检测结果显示,HE染色见MG组有重度脂肪变性、大小不等的脂滴弥漫沉积在胞浆,油红染色呈现染成红色的脂滴大量沉积,呈重度脂肪肝变,提示造模成功; CG组、BJ组、BJB组肝组织脂质沉积较MG组、SI组、BJSI组及BJBSI组肝组织明显减少,肝损害程度减轻; BJ组、BJB组血清TG、TC、LDL-c、MDA、ALT及AST较MG组、SI组、BJSI组和BJBSI组降低,HDL-c、SOD、GSH和SIRT1蛋白水平较MG组、SI组、BJSI组和BJBSI组升高(P <0. 05),且BJB组变化大于BJ组、BJBSI组变化小于BJSI组,差异有统计学意义(P <0. 05)。结论:蓝莓及蓝莓联合益生菌具有抗氧化活性,能拮抗酒精诱导的脂代谢紊乱和脂质过氧化,减轻氧化应激,从而减轻AFLD肝损伤,其机制可能与上调小鼠肝脏SIRT1的表达有关。
Objective: To analyze the protective effects and possible pathogenesis of dietary blueberry and blueberry combined with probiotics on alcoholic fatty liver disease(AFLD). Methods: 58 C57 BL/6 J rats were divided into normal control group(CG group),model group(MG group),blueberry juice group(BJ group),blueberry juice combined probiotics group(BJB group),SIRT1 siRNA blocking group(SI group). Blueberry juice + SIRT1 blocking group(BJSI group) and blueberry juice combined probiotics + SIRT1 blocking group(BJBSI group). There were 10 rats in MG group and 8 rats in each other groups. In addition to the CG group,the other 6 groups were fed DeCarli '82 diet to make AFLD model(all rats were fed normal diet after the model was successfully made). the BJ group and BJB group were fed with blueberry and blueberry plus probiotics respectively 7 days before the model was made. After successful modeling,rats in the BJSI group and BJBSI group were treated with blueberry and blueberry combined with probiotics respectively,and LV-mediated SIRT1 siRNA was injected into abdominal liver of rats in the two groups. After 12 weeks of feeding,the success of modeling was confirmed by pathological methods. The blood was taken from the eye frame of each group 10 days after the model was made,and the liver tissue was taken after death caused by cervical vertebrae dislocation. The histological changes of the liver in each group were observed by pathological method. Triglyceride(TG),total cholesterol(TC),alanine aminotransferase(ALT),aspartate aminotrans-ferase(AST),low-density lipoprotein-cholesterol(LDL-c) and high-density lipoprotein-cholesterol(HDLc) were determined by biochemical method. Malondialdehyde(MDA),reduced glutathione(GSH)levels,superoxide dismutase(SOD) levels and expressions of silent information regulator 1(SIRT1)were tested by ELISA. Results: Pathological examination indicated that HE staining showed severe steatosis and diffusion and deposit of lipid droplets of varying sizes in the cytoplasm in MG group; oil red staining showed a large amount of deposition of fat droplets dyed red and severe fatty liver degeneration,which demonstrated that the model was successfully established. The lipid accumulation were significantly lower and the degree of liver damage were reduced in CG,BJ and BJB groups than that in MG,SI,BJSI and BJBSI groups. Serum TG,TC,LDL-c,MDA,ALT and AST in BJ group and BJB group were lower than those in MG group,SI group,BJSI group and BJBSI group,but HDL-c,SOD,GSH and SIRT1 protein were higher than those in MG group,SI group,BJSI group and BJBSI group(P < 0. 05). The change of BJB group was greater than that of BJ group,and the change of BJBSI group was smaller than that of BJSI group; the differences were statically significant(P < 0. 05).Conclusions: Blueberry and blueberry combined probiotics have antioxidant activity,can antagonize lipid metabolism disorder and lipid peroxidation induced by alcohol,alleviate oxidative stress,and reduce AFLD liver injury. The pathogenesis may be related to up-regulation of SIRT1 expression in rats' livers.

关键词(KeyWords)： 蓝莓;益生菌;酒精性脂肪性肝病;氧化应激;脂质过氧化;沉默信息因子1
blueberry;probiotics;alcoholic fatty liver disease;oxidativestress;lipid peroxidation;silent information regulator 1

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金地区基金资助项目(81560100)

作者(Author): 沈艳艳,任婷婷
SHEN Yanyan,REN Tingting

DOI: 10.19367/j.cnki.1000-2707.2018.11.001

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