白藜芦醇通过糖代谢重编程抑制DEN诱导大鼠肝细胞癌前阶段的恶性增殖  

Resveratrol Inhibits DEN-induced Malignant Proliferation of Rat Hepatocellular Precancerous Stage Through Reprogramming of Glucose Metabolism

作　　者：齐晓茹 孙思语 程金妞 赵安妮 刘晓梅 刘真真 那莎[1] 李璐[1,2] QI Xiao-Ru;SUN Si-Yu;CHENG Jin-Niu;ZHAO An-Ni;LIU Xiao-Mei;LIU Zhen-Zhen;NA Sha;LI Lu(Department of Biochemistry,Department of Biological Sciences,College of Integrated Traditional Chinese and Western Medicine,Anhui University of Traditional Chinese Medicine,Hefei 230012,China;Institute of Integrated Traditional Chinese and Western Medicine,Anhui Academy of Traditional Chinese Medicine,Hefei 230012,China)

机构地区：[1]安徽中医药大学中西医结合学院生物科学系生物化学教研室,合肥230012 [2]安徽省中医药科学院中西医结合研究所,合肥230012

出　　处：《中国生物化学与分子生物学报》2022年第9期1213-1225,共13页Chinese Journal of Biochemistry and Molecular Biology

基　　金：安徽省自然科学基金项目(No.1808085MH301);国家自然科学基金项目(No.81703826)资助。

摘　　要：白藜芦醇(resveratrol,RES)可抑制肝癌细胞的生长与增殖。但其在癌前阶段的作用尚不十分清楚。本文研究白藜芦醇对二乙基亚硝胺(diethylinitrosamine,DEN)诱导大鼠肝癌前阶段的作用及机制。SD大鼠分为正常对照组、RES处理组、DEN处理组和RES-DEN处理组。研究结果表明,DEN处理大鼠8周时,肝细胞的总增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)升高至2倍(P<0.05),核内PCNA蛋白表达水平升高至3倍(P<0.001),而RES-DEN处理组大鼠肝细胞总PCNA(P<0.05)和核内PCNA蛋白表达水平(P<0.001)降低。结果提示,RES可显著抑制肝细胞恶性增生。通过非靶向代谢物组学及代谢通路富集分析,结果表明,RES-DEN处理大鼠的肝细胞中,虽然磷酸戊糖途径向糖酵解途径的转变增强,但相较于DEN组大鼠,糖酵解水平并未出现显著提高,提示磷酸烯醇式丙酮酸-丙酮酸-乳酸这条代谢途径被抑制。进一步验证发现,这条代谢途径上的关键酶M2型丙酮酸激酶(M2-type pyruvate kinase,PKM2)和乳酸脱氢酶(lactate dehydrogenase,LDHA)蛋白质表达水平被抑制(P<0.05)。RES可通过调节糖代谢重编程,在肝癌的癌前阶段抑制DEN诱导的大鼠肝细胞的过度增殖,为RES预防肝癌提供了实验依据。Resveratrol(RES)can inhibit the growth and proliferation of liver cancer cells.However,its role in the precancerous stage is still unclear.This paper aims to study the effect and mechanism of RES on the precancerous stage of liver cancer in rats induced by diethylinitrosamine(DEN).SD rats were divided into normal control group,RES treatment group,DEN treatment group and RES-DEN treatment group.The results showed that after the rats were treated with DEN for 8 weeks,the total expression level of proliferating cell nuclear antigen(PCNA)of hepatocytes increased to 2-fold(P<0.05),and the expression level of PCNA protein in the nucleus increased to 3-fold(P<0.001).However,the expression levels of total PCNA(P<0.05)and nuclear PCNA protein(P<0.001)in hepatocytes of rats treated with RES-DEN decreased,suggesting that RES could significantly inhibit the liver malignant proliferation of cells.Through non-targeted metabolomics and KEGG metabolic pathway enrichment analysis,the results showed that the level of glycolysis did not increase significantly in the hepatocytes of RES-DEN-treated rats,although the transition from the pentose phosphate pathway to the glycolysis pathway was enhanced when compared with the DEN group rats.This finding suggested that the metabolic pathway of phosphoenolpyruvate-pyruvate-lactate was inhibited.Further verification found that the protein expression levels of key enzymes M2-type pyruvate kinase(PKM2)and lactate dehydrogenase(LDHA)in this metabolic pathway were inhibited(P<0.05).RES can reprogram glucose metabolism and inhibit DEN-induced excessive proliferation of rat hepatocytes in the precancerous stage of liver cancer,providing an experimental basis for RES to prevent liver cancer.

关 键 词：白藜芦醇 二乙基亚硝胺 糖代谢重编程 癌前期 代谢物组学 

分 类 号：Q591.4[生物学—生物化学]