甲状旁腺激素相关蛋白加重蛋氨酸胆碱缺乏饲料诱导的小鼠非酒精性脂肪性肝病进展  

Parathyroid hormone-related protein aggravates nonalcoholic fatty liver disease induced by methionine choline-deficient diet in mice

作　　者：覃碧艳 刁娜 白岚[1] QIN Biyan;DIAO Na;BAI Lan(Guangdong Provincial Key Laboratory of Gastroenterology,Department of Gastroenterology,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China;Guangdong Provincial Key Laboratory of Colorectal Diseases,Department of Gastroenterology,Sixth Affiliated Hospital of Sun Yat-sen University,Guangzhou 510655,China)

机构地区：[1]南方医科大学南方医院消化内科广东省胃肠疾病重点实验室,广东广州510515 [2]中山大学附属第六医院消化内科//广东省结直肠盆底疾病重点实验室,广东广州510655

出　　处：《南方医科大学学报》2021年第7期1037-1043,共7页Journal of Southern Medical University

基　　金：国家自然科学基金(81470790);广东省科技计划项目(2017B020209003);国家卫生计生委公益事业专项(201502026);中国博士后科学基金(2019M663286)。

摘　　要：目的研究甲状旁腺激素相关蛋白(PTHr P)对蛋氨酸胆碱缺乏饲料(MCD)诱导的小鼠非酒精性脂肪性肝病(NAFLD)的影响。方法构建小鼠NAFLD模型并分为空白对照组(普通饲料喂养4周)、AAV-Vehicle组(注射空载腺相关病毒AAVVehicle)和AAV-PTHr P组(注射PTHr P过表达腺相关病毒)。实验期间监测小鼠体质量。收集小鼠肝脏组织进行HE染色、油红染色和天狼星红染色评估肝脏组织病理学改变;检测肝脏及血清中谷草转氨酶、谷丙转氨酶、甘油三酯和游离脂肪酸浓度评估小鼠肝脏损伤及脂肪代谢水平。250μmol/LFFAs诱导小鼠正常肝细胞和人正常肝细胞24 h构建NAFLD细胞。根据有无PTHr P处理分为PTHr P组、对照组。油红及尼罗红染色检测细胞内脂质沉积程度;CCK8试剂盒检测PTHr P对脂肪变性肝细胞的毒性作用。结果动物实验:对比AAV-Vehicle组,AAV-PTHr P组小鼠体重下降更为快速;AAV-PTHr P组小鼠肝脏谷草转氨酶(P<0.05)、谷丙转氨酶(P<0.05)、甘油三酯(P<0.01)、游离脂肪酸(P<0.05)水平,NAS评分以及SAF评分均显著升高(P<0.01,P<0.05),肝脏脂滴堆积更为明显。细胞实验:同时加入PTHr P,小鼠及人NAFLD细胞模型的脂质沉积更为明显(P<0.01),且细胞活性下降(P<0.05)。结论PTHr P可能通过促进肝细胞脂滴沉积,加重MCD诱导的小鼠NAFLD。Objective To study the effect of parathyroid hormone-related protein(PTHr P)on nonalcoholic fatty liver disease(NAFLD)induced by methionine choline-deficient diet(MCD)in mice.Methods Twelve male C57 BL/6 J mice were randomized into blank control group,vehicle group and PTHr P group(n=4).The mice in vehicle group and PTHr P group received injections of a control adeno-associated virus(AAV)vector and an AVV vector carrying PTHr P(AAV-PTHr P)gene,respectively,followed one week later by MCD feeding for 3 weeks;the mice in the blank control were fed a normal diet for 4 weeks.Body weight changes of the mice were monitored during the experiment.At the end of the experiment,liver tissues were harvested from the mice for histological analysis using HE staining,oil red O staining,and Sirius red staining.The levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),triglyceride,and free fatty acids(FFAs)in the liver and serum were detected to assess hepatic impairment and lipid metabolism of the mice.Cell models of NAFLD were established in mouse and human normal liver cells by treatment with 250μmol/L FFAs for 24 h,and the effect of AAV-PTHr P on lipid deposition and viability of the cells were tested using Oil Red O and Nile red staining and CCK8 assay.Results Treatment with AAV-PTHr P,as compared with the control AVV vector,caused more rapid reduction of body weight in mice with MCD feeding and significantly increased the levels of AST(P<0.05),ALT(P<0.05),triglyceride(P<0.01)and FFA(P<0.05)in the liver and the scores of NAS(P<0.01)and SAF(P<0.05).HE and Oil red O staining of the liver tissue revealed obvious lipid deposition after MCD feeding,which was more serious in PTHr P group.In the cell experiment,FFAs induced steatosis in both mouse and human hepatocytes,and treatment with PTHr P increased the accumulation of lipid droplets and lowered the viability of the cell model of NAFLD(P<0.01 or 0.05).Conclusion PTHr P may aggravate MCD-induced NAFLD in mice by promoting the deposition of lipid droplets in th

关 键 词：非酒精性脂肪性肝病 甲状旁腺激素相关蛋白 蛋氨酸胆碱缺乏饲料 

分 类 号：R575.5[医药卫生—消化系统]