大黄素甲醚改善小鼠非酒精性脂肪性肝病的研究

The improvement provided by physcion on non-alcoholic fatty liver disease in mice

大黄素甲醚(physcion,PHY)是一种来源于大黄等中药的蒽醌类化合物。本实验旨在探究PHY对非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)的改善作用及其机制。采用蛋氨酸胆碱缺乏饲料(methionine-and choline-deficient diet,MCD)喂养6周诱导小鼠NAFLD疾病模型(本实验获得上海中医药大学实验动物伦理委员会批准,批准号:PZSHUTCM190705019)。结果显示,PHY(5和20 mg·kg^(-1))能改善MCD诱导NAFLD小鼠的肝损伤,减轻肝脏脂肪累积,降低升高的NAFLD活动性评分(NAFLD activity score,NAS)。Western blot及酶活力实验结果提示,PHY能提升肝脏及L-02细胞中肉毒碱棕榈酰基转移酶1A(carnitine palmitoyltransferase 1A,CPT1A)蛋白表达及酶活力,但real-time PCR结果显示PHY不影响Cpt1a mRNA表达。免疫荧光结果提示,PHY(10和25μmol·L^(-1))能缓解游离脂肪酸(non-esterified fatty acids,NEFA)诱导的人正常肝L-02细胞中的线粒体损伤。Seahorse实验结果提示,PHY给药后能提升L-02细胞内线粒体基础呼吸能力、最大呼吸能力、ATP合成和储备呼吸能力,但不影响线粒体质子漏。以上研究表明,PHY可能通过改善线粒体功能,促进脂肪酸β氧化,减少肝脏中脂肪的堆积,发挥改善NAFLD的药效。

Physcion(PHY)is an anthraquinone compound derived from traditional Chinese medicine such as Rhei Radix et Rhizoma.The aim of this study is to investigate the improvement of PHY on non-alcoholic fatty liver disease(NAFLD)and its underlying mechanism.NAFLD was induced in mice by feeding with the methionine-and choline-deficient diet(MCD)for 6 weeks.This experiment was approved by the Experimental Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine(approval number:PZSHUTCM190705019).The results displayed that PHY(5 and 20 mg·kg^(-1))reversed liver damage,reduced hepatic lipid accumulation and decreased the elevated NAFLD activity score(NAS)in MCD-fed NAFLD mice.Results from Western blot and enzyme activity demonstrated that PHY could enhance the protein expression and enzyme activity of carnitine palmitoyltransferase 1A(CPT1A)in the liver and L-02 cells,but it did not affect Cpt1a mRNA expression.Immunofluorescence results indicated that PHY(10 and 25μmol·L^(-1))could reduce the mitochondrial injury induced by non-esterified fatty acids(NEFA)in L-02 cells.Results from seahorse assay showed that PHY could enhance mitochondrial basic respiration,maximal respiration,ATP synthesis and reserve respiration in L-02 cells treated with NEFA,but had no effect on mitochondrial proton leakage.In summary,PHY reversed mitochondrial damage and enhanced fatty acidβ-oxidation,thereby reducing hepatic steatosis and improving NAFLD.