摘要
基于Nrf2/HO-1铁死亡信号通路探讨补阳还五汤对血管紧张素Ⅱ(Angiotensin Ⅱ,AngⅡ)诱导的H9c2大鼠心肌细胞铁死亡的影响。体外培养H9c2心肌细胞,分别设正常组(ZC)、模型组(MX)、补阳还五汤低剂量组(BD)、补阳还五汤中剂量组(BZ)、补阳还五汤高剂量组(BG)和阳性对照组(YX)。除正常组外,其余各组采用1μmol/L AngⅡ建立心肌细胞损伤模型,同时补阳还五汤低、中、高剂量组分别用含0.5、1、2 mg/mL补阳还五汤的培养基培养,阳性对照组使用含10μmol/L铁死亡抑制剂Ferrostatin-1(Fer-1)的培养基培养,24 h后收集细胞。采用CCK-8法检测各组心肌细胞活性;细胞铁含量检测试剂盒测定细胞内铁含量;免疫荧光染色法检测各组Nrf2蛋白表达;q-PCR法检测各组细胞核转录相关因子2(Nrf2)、血红加氧酶-1(HO-1)、谷胱甘肽过氧化物酶4(GPX4)的mRNA表达水平;ELISA试剂盒检测细胞谷胱甘肽(GSH)和GPX4的蛋白表达水平;Western blot法检测Nrf2和HO-1的蛋白表达。结果表明,补阳还五汤高剂量组可明显提高AngⅡ诱导的H9c2细胞活力下降(P<0.01),减少AngⅡ条件下铁含量(P<0.05);与模型组相比,补阳还五汤高剂量组能显著逆转AngⅡ诱导的H9c2细胞GSH、GPX4、Nrf2和HO-1的基因和蛋白表达水平的下降,使AngⅡ条件下受到抑制的Nrf2表达增加,与阳性对照组结果一致(P<0.01,P<0.05)。补阳还五汤可能通过Nrf2/HO-1信号通路调控心肌细胞铁死亡从而保护高血压继发的心肌损伤。
To investigate the regulatory effect of Buyang Huanwu Decoction on ferroptosis of H9c2 rat cardiomyocytes in the environment of angiotensin Ⅱ(AngⅡ)which based on Nrf2/HO-1 signaling pathway.Rat myocardial cells(H9c2)were cultured in vitro,and the normal group(ZC),model group(MX),low dosage group of Buyang Huanwu Decoction(BD),medium dosage group of Buyang Huanwu Decoction(BZ),high dosage group of Buyang Huanwu Decoction(BG)and positive control group(YX)were set up.All groups were treated with 1μmol/L AngII established a myocardial cell injury model except for the normal group,the intervention group of traditional Chinese medicine was respectively cultured in culture media containing 0.5,1 and 2 mg/mL Buyang Huanwu decoction,the YX was cultured with medium containing 10μmol/L ferrostatin-1(Fer-1),and the cells were collected after 24 h.The activity of H9c2 cells in each group was assessed by CCK-8 assay.Cytometric iron content detection kit to determine the iron content of H9c2 cells.Immunofluorescence staining to detect the expression of Nrf2 protein in each group.The mRNA expression levels of nuclear transcription-related factor 2(Nrf2),hemored oxygenase-1(HO-1),and glutathione peroxidase 4(GPX4)were detected by q-PCR method in each group.ELISA kits was used to detect the protein expression levels of glutathione(GSH)and GPX4 in H9c2 cells.The Western blot method was used to detect the protein expression of Nrf2 and HO-1 in H9c2 cells.The results show that the high-dose groups of Buyang Huanwu Decoction significantly increases AngⅡ-induced decrease in H9c2 cells viability(P<0.01),and reduced iron content under AngⅡ condition(P<0.05).Compared with the model group,the BG significantly reversed the decline of gene and protein expression levels of GSH,GPX4,Nrf2 and HO-1 in H9c2 cells induced by AngⅡ,and increases the expression of Nrf2,which was inhibited under AngII conditions,consistent with the positive control results(P<0.01,P<0.05).Buyang Huanwu Decoction may regulate H9c2 cells ferroptosis through N
作者
薛亚楠
王建波
曲怡
高佳馨
张云雨
张立德
XUE Ya-nan;WANG Jian-bo;QU Yi;GAO Jia-xin;ZHANG Yun-yu;ZHANG Li-de(Innovative Engineering Technology Center of Traditional Chinese Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications,Shenyang 110847,China;Graduate School of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)
出处
《科学技术与工程》
北大核心
2024年第1期155-162,共8页
Science Technology and Engineering
基金
辽宁中医药大学自然科学课题(2100222080)
国家级大学生创新创业训练项目(202210162006)
辽宁省教育厅育苗项目(L202025)。
关键词
补阳还五汤
心肌
铁死亡
高血压
Buyang Huanwu Decoction
cardiomyocytes
ferroptosis
hypertension
