摘要
目的探讨黄芪-葛根配伍对2型糖尿病(T2DM)胰岛素抵抗(IR)大鼠肝细胞铁死亡的影响及潜在机制。方法将雄性SD大鼠60只分为对照组(12只)与造模组(48只),造模组大鼠以高脂饲料连续喂养4周后按25 mg/kg的剂量一次性尾静脉注射1%链脲佐菌素溶液以复制T2DM IR大鼠模型。将造模成功的大鼠随机分为模型组、黄芪-葛根配伍组[QG组,4.05 g/(kg·d)灌胃]、铁死亡抑制剂ferrostatin-1组(Fer-1组,5 mg/kg隔日1次腹腔注射)、黄芪-葛根配伍+铁死亡诱导剂erastin组[QG+erastin组,4.05 g/(kg·d)灌胃+erastin 10 mg/(kg·d)腹腔注射]。干预4周后,检测各组大鼠血清空腹血糖(FBG)、空腹胰岛素(FINS)水平,并计算稳态模型胰岛素抵抗指数(HOMA-IR)和胰岛素敏感指数的自然对数(IAI);检测血清总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)水平,血清Fe^(2+)及Fe水平,肝脏组织中谷胱甘肽(GSH)、丙二醛(MDA)、超氧化物歧化酶(SOD)水平以及烟酰胺腺嘌呤二核苷酸磷酸(NADP+)/还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)和活性氧(ROS)水平;观察其肝脏组织病理形态,检测肝脏组织谷胱甘肽过氧化物酶4(GPX4)、铁蛋白重链1(FTH1)、长链脂酰辅酶A合成酶3(ACSL3)、ACSL4、线粒体铁蛋白(FTMT)、胱氨酸/谷氨酸反向转运蛋白(xCT)的蛋白表达情况。结果与对照组比较,模型组大鼠肝脏组织内肝细胞排列紊乱、肿胀,细胞核染色加深,可见较多炎症细胞浸润,大量肝细胞空泡及脂肪变性;FBG(给药后)、TC、TG、LDL-C、AST、ALT、FINS、MDA、ROS、Fe^(2+)、Fe水平及HOMA-IR,NADP+/NADPH和ACSL4蛋白的表达均显著升高或上调(P<0.01);HDL-C、GSH、SOD水平,IAI及GPX4、FTH1、ACSL3、FTMT、xCT蛋白的表达均显著降低或下调(P<0.01)。与模型组比较,QG组和Fer-1组大鼠肝脏组织病理损伤及上述指标水平均有不同程度改善,且大部分指标的变化差异有统计学意义(P<0.01或P<0.05)。与QG组比较,QG+erastin组大鼠上述指标水平的改善均被显著逆转(P<0.01)。结论黄芪-葛根配伍水煎液可降低T2DM IR大鼠FBG水平,减轻其IR程度和肝脏组织铁负荷,缓解其肝脏组织病理损伤,上述作用与其抑制铁死亡有关。
OBJECTIVE To explore the effect and potential mechanism of the compatibility of Astragali Radix-Puerariae Lobatae Radix on ferroptosis of liver cells in type 2 diabetes mellitus(T2DM)insulin resistance(IR)rats.METHODS Sixty male SD rats were randomly divided into control group(12 rats)and modeling group(48 rats).The modeling group was fed with a highfat diet for 4 consecutive weeks and then given a one-time tail vein injection of 1%streptozotocin to establish T2DM IR model.The model rats were randomly divided into model group,the compatibility of Astragali Radix-Puerariae Lobatae Radix group[QG group,4.05 g/(kg·d),intragastric administration],ferroptosis inhibitor ferrostatin-1 group[Fer-1 group,5 mg/kg by intraperitoneal injection,once every other day],the compatibility of Astragali Radix-Puerariae Lobatae Radix+ferroptosis inducer erastin group[QG+erastin group,4.05 g/(kg·d)by intragastric administration+erastin 10 mg/(kg·d),intraperitoneal injection].After 4 weeks of intervention,serum fasting blood glucose(FBG)and fasting insulin(FINS)were measured in each group of rats,and homeostasis model assessment of insulin resistance(HOMA-IR)and the natural logarithm of insulin action index(IAI)were calculated;the serum levels of total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),aspartate transaminase(AST)and alanine transaminase(ALT),Fe^(2+) and Fe content,glutathione(GSH),malondialdehyde(MDA)and superoxide dismutase(SOD)levels,NADP+/NADPH ratio and reactive oxygen species(ROS)were determined.The pathological morphology of its liver tissue was observed;the protein expressions of glutathione peroxidase 4(GPX4),ferritin heavy chain 1(FTH1),long-chain acyl-CoA synthetase 3(ACSL3),ACSL4,ferritin mitochondrial(FTMT),and cystine/glutamate anti-porter(xCT)in the liver tissue of rats were detected.RESULTS Compared with control group,the liver cells in the model group of rats showed disordered arrangement,swelling,deepened nuclear staining,and more infiltration of inflammatory cells,as well as a large number of hepatocyte vacuoles and steatosis;FBG(after medication),the levels of TC,TG,LDL-C,AST,ALT,FINS,MDA and ROS,HOMA-IR,Fe^(2+) and Fe content,NADP+/NADPH ratio and protein expression of ACSL4 were significantly increased or up-regulated,while the levels of HDL-C,GSH and SOD,IAI,protein expressions of GPX4,FTH1,ACSL3,FTMT and xCT were significantly reduced or down-regulated(P<0.01).Compared with the model group,both QG group and Fer-1 group showed varying degrees of improvement in pathological damage of liver tissue and the levels of the above indicators,the differences in the changes of most indicators were statistically significant(P<0.01 or P<0.05).Compared with QG group,the improvement of the above indexes of QG+erastin group had been reversed significantly(P<0.01).CONCLUSIONS The compatibility decoction of Astragali Radix-Puerariae Lobatae Radix can reduce the level of FBG in T2DM IR rats,and alleviate IR degree,ion overload and pathological damage of liver tissue.The above effects are related to the inhibition of ferroptosis.
作者
魏爽
郝峰
章文春
赵张旸
李冀
韩东卫
邢欢
WEI Shuang;HAO Feng;ZHANG Wenchun;ZHAO Zhangyang;LI Ji;HAN Dongwei;XING Huan(College of Traditional Chinese Medicine,Jiangxi University of Chinese Medicine,Nanchang 330004,China;School of Clinical Medicine,Jiangxi University of Chinese Medicine,Nanchang 330004,China;School of Basic Medicine,Heilongjiang University of Chinese Medicine,Harbin 150040,China)
出处
《中国药房》
北大核心
2025年第1期57-63,共7页
China Pharmacy
基金
国家自然科学基金项目(No.81904090,No.81874426)
江西省自然科学基金-基础研究计划项目(No.20242BAB-20443)
江西中医药大学博士科研启动基金项目(No.2024BSZR027)。
关键词
黄芪
葛根
配伍
铁死亡
2型糖尿病
胰岛素抵抗
Astragali Radix
Puerariae Lobatae Radix
compatibility
ferroptosis
type 2 diabetes mellitus
insulin resistance
作者简介
第一作者:魏爽,副教授,博士。研究方向:方剂配伍。E-mail:864031535@qq.com;通信作者:邢欢,副教授,博士。研究方向:中医病因病机和基础。E-mail:xinghuan@jxutcm.edu.cn。
