摘要
目的:基于甲状腺肿大模型,探究海藻甘草反药组合在不同剂量条件下是否会对模型大鼠产生肝脏毒性,表现出“相反”作用,并探讨其对模型大鼠肝脏蛋白的合成功能及相关机制的影响。方法:将90只Wistar雄性大鼠随机分为空白组、模型组、阳性药(优甲乐)组(20μg·kg^(-1))、海藻甘草低剂量组(HG-D组,1.98 g·kg^(-1))、海藻甘草中剂量组(HG-Z组,3.96 g·kg^(-1))和海藻甘草高剂量组(HG-G组,7.92 g·kg^(-1)),共6组,空白组用去离子水灌胃,其余各组灌服丙硫氧嘧啶(PTU)复制甲状腺肿大病理模型,模型复制成功后,阳性药组灌服优甲乐14 d,各中药组灌服相应剂量海藻甘草药液14 d,末次给药12 h后进行取材,检测各组大鼠血清天门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、碱性磷酸酶(ALP)、总蛋白(TP)、白蛋白(ALB)的含量;苏木素-伊红(HE)、马松(Masson)染色观察肝脏病理学变化;实时荧光定量聚合酶链式反应(qPCR)检测肝脏组织蛋白激酶B(AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)、真核翻译起始因子4E结合蛋白1(4EBP1)、真核翻译起始因子4E(eIF4E)mRNA的表达情况,蛋白免疫印迹法(WB)检测肝脏组织蛋白合成信号通路AKT、mTOR、pmTOR、4EBP1蛋白表达情况。结果:与空白组比较,模型组ALT水平显著提高(P<0.05),ALB、AST/ALT水平显著下降(P<0.05,P<0.01),AKT、mTOR、4EBP1、eIF4E mRNA表达显著降低(P<0.05,P<0.01),AKT、mTOR、pmTOR、4EBP1蛋白表达显著降低(P<0.01);与模型组比较,HG-D组TP、ALB水平显著上升(P<0.05,P<0.01),AKT、mTOR、4EBP1、eIF4E mRNA表达显著上升(P<0.05,P<0.01),AKT、mTOR、pmTOR、4EBP1蛋白表达显著上升(P<0.01);与HG-D组相比,HG-Z组、HG-G组pmTOR水平显著降低(P<0.05)。结论:海藻甘草反药组合在不同剂量条件下对甲状腺肿大模型大鼠肝脏未产生明显毒性表现“相反”作用,而是对蛋白合成功能有不同程度的保护效果,其中HG-D组的保护效果最好,优于HG-Z组和HG-G组,其作用机制可能与激活AKT/mTOR/4EBP1信号通路有关。
Objective:To explore whether the seaweed-glycyrrhiza reverse drug combination causes hepatic toxicity and exhibits“opposite”effects on thyroid goiter model rats under different dosage conditions,and to investigate its impact on the hepatic protein synthesis function and related mechanisms.Methods:Ninety male Wistar rats were randomly divided into six groups:control group,model group,positive drug(Euthyrox)group(20μg·kg^(-1)),low-dose(1.98 g·kg^(-1)),medium-dose(3.96 g·kg^(-1)),and high-dose seaweed-glycyrrhiza(HG-G)group(7.92 g·kg^(-1)),with 15 rats per group.The control group was treated with deionized water by gavage.The other groups were treated with propylthiouracil(PTU)to replicate the thyroid goiter model.After successful model induction,the positive drug group was treated with Euthyrox for 14 days,while the seaweed-glycyrrhiza groups were treated with the respective doses of seaweed-glycyrrhiza solution for 14 days.After the last administration,rats were sacrificed 12 hours later.Serum levels of AST,ALT,ALP,total protein(TP),and albumin(ALB)were measured.Histopathological changes in the liver were observed using hematoxylin-eosin(HE)and Masson staining.mRNA expression of protein kinase B(Akt),mammalian target of rapamycin(mTOR),eukaryotic translation initiation factor 4E binding protein 1(4EBP1),and eukaryotic translation initiation factor 4E(eIF4E)in liver tissue was measured by quantitative real-time PCR(qPCR).The protein expression levels of AKT,mTOR,p-mTOR,and 4EBP1 were detected by Western blotting.Results:Compared with the control group,the model group showed significantly increased ALT levels(P<0.05),and decreased ALB,AST/ALT ratios(P<0.05,P<0.01).The mRNA expressions of AKT,mTOR,4EBP1,and eIF4E were significantly reduced(P<0.05,P<0.01),and the protein expressions of AKT,mTOR,p-mTOR,and 4EBP1 were significantly decreased(P<0.01).Compared with the model group,the HG-D group showed significantly increased levels of TP and ALB(P<0.05,P<0.01),with significantly increased mRNA expression of AKT,mTOR,4EBP1,and eIF4E(P<0.05,P<0.01),and significantly increased protein expression of AKT,mTOR,p-mTOR,and 4EBP1(P<0.01).Compared with the HG-D group,the HG-Z and HG-G groups showed significantly lower p-mTOR levels(P<0.05).Conclusion:The seaweed-glycyrrhiza reverse drug combination did not exhibit obvious toxicity or“opposite”effects on the liver in the thyroid goiter model rats.Instead,it provided varying degrees of protection to hepatic protein synthesis function.The HG-D group showed the best protective effect,superior to the HG-Z and HG-G groups.The mechanism may be related to the activation of the AKT/mTOR/4EBP1 signaling pathway.
作者
徐向楠
吴美晶
董肖
田颐
廖文勇
刘晓庆
曹灿
于雪
范盎然
修琳琳
陈绍红
柳海艳
钟赣生
XU Xiangnan;WU Meijing;DONG Xiao;TIAN Yi;LIAO Wenyong;LIU Xiaoqing;CAO Can;YU Xue;FAN Angran;XIU Linlin;CHEN Shaohong;LIU Haiyan;ZHONG Gansheng(Beijing University of Chinese Medicine,Beijing 100029,China)
出处
《中医药学报》
2025年第5期9-17,共9页
Acta Chinese Medicine and Pharmacology
基金
国家自然科学基金项目(81973496)
北京中医药大学纵向科研发展基金项目(2024-ZXFZJJ-JW-056)。
