海藻甘草反药组合在海藻玉壶汤中加减应用对甲状腺肿大模型大鼠肝脏氧化应激的影响  

Effect of Sargassum and Glycyrrhizae Radix et Rhizoma Incompatible Pair with Haizao Yuhutang on Oxidative Stress in Liver of Goiter Rats

作　　者：董肖 田颐 刘晓庆 曹灿[1] 廖文勇 徐向楠 吴美晶 柳海艳[1] 陈绍红[1] 于雪[1] 范盎然[1] 修琳琳[1] 钟赣生[1] DONG Xiao;TIAN Yi;LIU Xiaoqing;CAO Can;LIAO Wenyong;XU Xiangnan;WU Meijing;LIU Haiyan;CHEN Shaohong;YU Xue;FAN Angran;XIU Linlin;ZHONG Gansheng(Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学,北京100029

出　　处：《中国实验方剂学杂志》2024年第14期37-45,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82274118)。

摘　　要：目的:观察海藻和甘草反药组合在2020年版《中华人民共和国药典》高限2倍剂量条件下于海藻玉壶汤中加减配伍应用对甲状腺肿大模型大鼠肝脏氧化应激的影响。方法:将128只雄性Wistar大鼠随机分为空白组、模型组、优甲乐组(20μg·kg^(-1))、海藻玉壶汤(HYT)组(12.06 g·kg^(-1))、HYT去海藻(HYT-H)组(9.90 g·kg^(-1))、HYT去甘草(HYT-G)组(10.26 g·kg^(-1))、HYT去海藻甘草(HYT-HG)组(8.10 g·kg^(-1))、海藻甘草(HG)组(3.96 g·kg^(-1)),空白组给予去离子水灌胃,其余各组灌胃丙硫氧嘧啶(PTU)复制甲状腺肿大病理模型,以优甲乐作为阳性药,其余各中药组给予相应药液灌胃,在末次给药后12 h进行取材。检测各组大鼠血清天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、碱性磷酸酶(ALP)水平;肝脏组织中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)、活性氧(ROS)的水平;苏木素-伊红(HE)染色观察肝脏病理学变化;实时荧光定量聚合酶链式反应(Real-time PCR)检测肝组织Kelch样ECH相关蛋白1(Keap1)、核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)、p53、胱天蛋白酶-3(Caspase-3)mRNA表达情况;蛋白免疫印迹法(Western blot)检测肝组织氧化应激相关信号通路Nrf2、HO-1蛋白表达情况。结果:与正常组比较,模型组大鼠血清ALT水平、肝脏组织中MDA和ROS含量明显升高(P<0.05,P<0.01),SOD、GSH-Px活性显著降低(P<0.01),Keap1 mRNA表达显著升高(P<0.01),Nrf2、HO-1 mRNA及蛋白表达明显降低(P<0.05,P<0.01);与模型组比较,HYT组大鼠血清AST、ALT、ALP水平明显降低(P<0.05,P<0.01),肝脏组织中MDA和ROS含量显著降低(P<0.01),SOD、GSH-Px活性显著升高(P<0.01),Keap1、p53、Caspase-3 mRNA表达显著降低(P<0.01),Nrf2、HO-1 mRNA及蛋白表达明显升高(P<0.05,P<0.01)。结论:在2020年版《中华人民共和国药典》高限2倍剂量条件下,海藻甘草反药组合于HYT中加减配伍应用对甲状�Objective:To observe the effects of Sargassum and Glycyrrhizae Radix et Rhizoma incompatible pair with the Haizao Yuhutang(HYT)on oxidative stress in the liver of goiter rats under the condition of 2 times the dose limit of the Pharmacopoeia of the People's Republic of China 2020.Method:A total of 128 male Wistar rats were randomly divided into a blank group,a model group,a euthyrox group(20μg·kg^(-1)),a HYT group(12.06 g·kg^(-1)),a HYT without Sargassum(HYT-H)group(9.90 g·kg^(-1)),a HYT without Glycyrrhizae Radix et Rhizoma(HYT-G)group(10.26 g·kg^(-1)),a HYT without Sargassum and Glycyrrhizae Radix et Rhizoma(HYT-HG)group(8.10 g·kg^(-1)),and a Sargassum and Glycyrrhizae Radix et Rhizoma(HG)group(3.96 g·kg^(-1)).The blank group was given deionized water by gavage,and the others were given propylthiouracil(PTU)to replicate the goiter pathological model.Euthyrox was taken as a positive control drug,and the rest of the Chinese medicine groups were given the corresponding decoction by gavage,the material was collected 12 hours after the last dose.The serum levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),alkaline phosphatase(ALP),and the contents of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),malondialdehyde(MDA),reactive oxygen species(ROS)in liver tissue were detected in each group.The pathological changes in the liver were observed via hematoxylin-eosin(HE)staining.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was utilized to detect the mRNA expressions of Kelch-like Ech-associated protein 1(Keap1),nuclear factor E2-related factor 2(Nrf2),heme oxygenase-1(HO-1),p53 and Caspase-3 in liver tissues.Western blot was adopted to detect the protein expressions of Nrf2 and HO-1 in liver tissues in oxidative stress-related signaling pathways.Result:Compared with control group,the model group showed significantly increased serum ALT level and contents of MDA and ROS in liver tissues(P<0.05,P<0.01),significantly reduced activities of SOD and GSH-Px in t

关 键 词：海藻玉壶汤 甲状腺肿大 肝脏氧化应激 核因子E2相关因子2(Nrf2) p53 

分 类 号：R2-0[医药卫生—中医学] R33R289R581