大黄对二乙基亚硝胺致大鼠肝癌癌前损伤作用及机制  

Effect and Mechanism of Dahuang(Rhei Radix Et Rhizoma)on Promoting Diethylnitrosamine to Hepatic Precancerous Injury in the Rats

作　　者：程金妞 刘真真 齐晓茹 孙思语 刘晓梅 赵安妮 那莎[1] 刘岩岩[1] 李璐[1,2] CHENG Jinniu;LIU Zhenzhen;QI Xiaoru;SUN Siyu;LIU Xiaomei;ZHAO Anni;NA Sa;LI Yanyan;LI Lu(Anhui University of Traditional Chinese Medicine,Hefei 230012,Anhui,China;Institute of Integrated Chinese and Western Medicine,Anhui Academy of Chinese Medicine,Hefei 230012,Anhui,China)

机构地区：[1]安徽中医药大学,安徽合肥230012 [2]安徽省中医药科学院中西医结合研究所,安徽合肥230012

出　　处：《辽宁中医药大学学报》2023年第1期14-19,F0003,共7页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81703826);安徽省自然科学基金项目(1808085MH301)。

摘　　要：目的 探讨大黄(Rhubarb)对二乙基亚硝胺致大鼠肝癌癌前损伤的作用及机制。方法 将SD大鼠分为正常对照组、大黄处理组、DEN处理组和大黄+DEN处理组4组。采用酶活性检测谷丙转氨酶(SGPT)活性、HE染色观察各组肝组织病理变化、Western blot法检测细胞外调节蛋白激酶(extracellular regulated protein kinases,Erk1/2)、细胞外调节蛋白激酶磷酸化(p-Erk1/2)、缺氧诱导因子1α(hypoxia inducible factor-1α,HIF-1α)、丙酮酸激酶同工酶M2型(pyruvate kinase M2,PKM2)等蛋白的表达。结果 与正常对照组、大黄处理组、DEN处理组相比,大黄+DEN组血清SGPT活性显著升高。病理切片HE染色显示,正常对照组大鼠和大黄处理组大鼠的肝细胞形态正常。DEN处理组的大鼠中,肝细胞排列紊乱并呈玻璃体样病变。与DEN处理大鼠相比,大黄+DEN处理的大鼠肝组织病理学变化更严重。Western blot结果表明与其他组相比,大黄+DEN组大鼠肝组织中Erk1/2、p-Erk1/2、HIF-1α、PKM2等蛋白水平显著升高。使用白藜芦醇作为PKM2的抑制剂,Westrn blot结果显示,与大黄+DEN处理2组相比,白藜芦醇+大黄+DEN处理组大鼠肝组织中PKM2蛋白水平显著降低;HE染色结果表明白藜芦醇减弱了大黄联合DEN对肝损伤的促进作用。结论 单独使用大黄对肝脏无损害,但大黄与DEN联合使用可促进大鼠的肝损伤,这与PKM2介导的有氧糖酵解有关。Objective To investigate the effect and mechanism of Dahuang(Rhei Radix Et Rhizoma)on prehepatic carcinoma injury of diethylnitrosamine in rats. Methods The SD rats were divided into four groups:normal group,Dahuang(Rhei Radix Et Rhizoma)treatment group,DEN treatment group,and Dahuang(Rhei Radix Et Rhizoma)+DEN treatment group. Glutamic-pyruvic transaminase(SGPT)activity was measured by enzyme activity,liver histopathological changes in each group were observed by HE staining,and the expression of extracellular regulated protein kinases(Erk1/2),extracellular regulatory protein kinase phosphorylation level(p-Erk1/2),hypoxia inducible factor-1α(HIF-1α)and pyruvate kinase M2(PKM2)was detected by Western blot. Results Compared with the control groups,the Dahuang(Rhei Radix Et Rhizoma)treatment groups,and the model groups,the serum SGPT was significantly increased in the Dahuang(Rhei Radix Et Rhizoma)-DEN group. When HE staining of the pathological sections revealed a normal morphology of hepatocytes in the control and Dahuang(Rhei Radix Et Rhizoma)-treated groups. In the DEN-treated group,hepatocytes were disordered and showed vitreous lesions. Compared with DEN treatment group,the pathological changes of liver tissue were more serious in the Dahuang(Rhei Radix Et Rhizoma)-DEN-treated rats. The Western blot results showed that the protein levels of Erk1/2,p-Erk1/2,HIF-1α and PKM2 were significantly increased in the liver tissues of the Dahuang(Rhei Radix Et Rhizoma)-DEN group compared with other groups. Using resveratrol as an inhibitor of PKM2,the Westrn blot results showed that the protein levels of PKM2 were significantly debased in the liver tissues of Resveratrol-Dahuang(Rhei Radix Et Rhizoma)-DEN group compared with Dahuang(Rhei Radix Et Rhizoma)-DEN group 2;HE staining showed that resveratrol attenuated the promoting effect of Dahuang(Rhei Radix Et Rhizoma)combined with DEN on liver damage. Conclusion Dahuang(Rhei Radix Et Rhizoma)alone was not damaging to the liver,but Dahuang(Rhei Radix Et Rhizoma)in comb

关 键 词：大黄 二乙基亚硝胺 肝毒性 丙酮酸激酶同工酶M2型 有氧糖酵解 

分 类 号：R735.7[医药卫生—肿瘤]