腺相关病毒介导的肝脏脂蛋白酯酶表达对小鼠高三酰甘油血症性急性胰腺炎的治疗作用  

Therapeutic effects of adeno-associated virus-mediated hepatic lipoprotein lipase expression on hypertriglyceridemic acute pancreatitis mice

作　　者：许尧 袁晨晨 路国涛 董小岩 吴小兵 刘国庆 李百强 李维勤 Xu Yao;Yuan Chenchen;Lu Guotao;Dong Xiaoyan;Wu Xiaobing;Liu Guoqing;Li Baiqiang;Li Weiqin(Department of Critical Care Medicine,Eastern Theater General Hospital of the Chinese People's Liberation Army(Jinling Hospital,Affiliated Hospital of Medical School,Nanjing University),Nanjing 210000,China;Genecradle Therapeutics Inc,Beijing 100176,China)

机构地区：[1]中国人民解放军东部战区总医院(南京大学医学院附属金陵医院)重症医学科,南京210000 [2]北京锦篮基因科技有限公司,北京100176

出　　处：《中华胰腺病杂志》2025年第1期50-56,共7页Chinese Journal of Pancreatology

基　　金：国家自然科学基金(81870441、82070669、82200722)。

摘　　要：目的探讨腺相关病毒载体5(AAV5)介导的肝脏脂蛋白酯酶(LPL)表达对小鼠血清三酰甘油(TG)代谢和高三酰甘油血症性急性胰腺炎(HTG-AP)的治疗作用。方法10只C57BL/6雄性Lpl^(+/-)小鼠按数字表法随机分为Lpl^(+/-)小鼠对照组和Lpl^(+/-)小鼠基因治疗组,每组5只。Lpl^(+/-)对照组小鼠尾静脉注射AAV5为载体、搭载增强型绿色荧光蛋白(EGFP)基因的重组病毒载体(AAV5-EGFP);Lpl^(+/-)基因治疗组小鼠尾静脉注射AAV5为载体、搭载人源LPLS447X基因的重组病毒载体(AAV5-LPLS447X),注射后14、28、56 d行口服脂耐量实验。20只野生型ICR小鼠按数字表法随机分为对照组和基因治疗组,每组10只。ICR对照组小鼠尾静脉注射AAV5-EGFP,ICR基因治疗组小鼠尾静脉注射AAV5-LPLS447X,注射后14 d腹腔注射P407溶液(0.5 g/kg)和雨蛙素(200μg/kg)进行HTG-AP造模。采用酶标仪检测血清TG、TC、淀粉酶、脂肪酶水平及注射肝素后血浆LPL活性,ELISA法检测血浆LPL浓度;荧光定量聚合酶链式反应(qRT-PCR)检测Lpl^(+/-)小鼠基因肝脏、心脏、脂肪组织LPL mRNA表达量;免疫组织化学法检测ICR小鼠基因治疗后28 d肝脏组织LPL蛋白表达;苏木精-伊红染色观察胰腺病理学变化。结果相较于Lpl^(+/-)小鼠对照组,Lpl^(+/-)基因治疗组血清TG基线水平从21 d开始显著下降,灌胃橄榄油后血清TG水平上升幅度和峰值均显著低于对照组,且肝脏LPL mRNA表达水平显著升高(1.96±0.11比1.02±0.12),差异均有统计学意义(P值均<0.05)。相较于ICR小鼠对照组,ICR小鼠基因治疗组血清TG和TC水平显著降低,28 d肝素后血浆LPL活性显著升高(0.17±0.05 mEq/L·h^(-1)比0.06±0.02 mEqL·h^(-1)),差异均有统计学意义(P值均<0.05)。免疫组织化学检测结果显示,ICR基因治疗组小鼠肝脏肝细胞膜表面高表达LPL蛋白,且胰腺组织水肿、炎症浸润和腺泡细胞坏死均较对照组显著减轻。结论LPLS447X治疗可促进小鼠肝脏表达LPLObjectiveTo investigate the therapeutic effects of adeno-associated virus vector 5(AAV5)-mediated hepatic lipoprotein lipase(LPL)expression on serum triglyceride(TG)metabolism and hypertriglyceridemic acute pancreatitis(HTG-AP)in mice.MethodsTen male C57BL/6 Lpl^(+/-)mice were randomly divided into two groups by a random number table:the Lpl^(+/-)control group and the Lpl^(+/-)gene therapy group,with five mice in each group.The Lpl^(+/-)control group received a tail vein injection of AAV5 vector carrying the enhanced green fluorescent protein(EGFP)gene(AAV5-EGFP),while the Lpl^(+/-)gene therapy group received a tail vein injection of AAV5 vector carrying the human LPLS447X gene(AAV5-LPLS447X).Oral fat tolerance tests were performed at 14,28,and 56 days post-injection.Twenty wild-type ICR mice were randomly divided into a control group and a gene therapy group,with ten mice in each group.The ICR control group was injected with AAV5-EGFP,and the ICR gene therapy group was injected with AAV5-LPLS447X.Fourteen days after injection,the mice underwent intraperitoneal injection of P407 solution(0.5 g/kg)and caerulein(200μg/kg)to induce HTG-AP.Serum TG,total cholesterol(TC),amylase,lipase levels,and plasma LPL activity after heparin injection were measured by microplate reader.Plasma LPL concentration was measured using an enzyme-linked immunosorbent assay(ELISA).LPL mRNA expression levels in the liver,heart,and adipose tissue of Lpl^(+/-)mice were determined by quantitative reverse-transcription polymerase chain reaction(qRT-PCR).LPL protein expression in the liver tissue of ICR mice was detected by immunohistochemistry at 28 days after gene therapy.Histopathological changes in the pancreas were observed using hematoxylin-eosin staining.ResultsCompared to the Lpl^(+/-)control group,the Lpl^(+/-)gene therapy group showed a significant decrease in serum TG levels starting from day 21.After oral administration of olive oil,the increase and peak of serum TG levels were significantly lower than those in the control group.Fu

关 键 词：脂蛋白脂肪酶 高三酰甘油血症 胰腺炎 腺病毒 病毒载体 基因治疗 

分 类 号：R73[医药卫生—肿瘤]