Zhuangtongyin Modulates Ferroptosis via the Nrf2-SCL7A11/xCT-Gpx4 Pathway to Improve Cerebral Ischemia-reperfusion Injury

WANG Chengyi; CAI Yuefang; NING Zhenqiu; DENG Minzhen; SUN Jingbo; CHUNG Sookja Kim; LI Yan; CHENG Xiao

doi:10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.20240701.001

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Zhuangtongyin Modulates Ferroptosis via the Nrf2-SCL7A11/xCT-Gpx4 Pathway to Improve Cerebral Ischemia-reperfusion Injury

Preclinic Research | 更新时间：2024-07-30

- Zhuangtongyin Modulates Ferroptosis via the Nrf2-SCL7A11/xCT-Gpx4 Pathway to Improve Cerebral Ischemia-reperfusion Injury
- Journal of Sun Yat-sen University(Medical Sciences) Vol. 45, Issue 4, Pages: 539-548(2024)
- 作者机构：
  
  1.广州中医药大学第二临床医学院，广东广州 510006
  2.深圳市人民医院//南方科技大学第一附属医院//暨南大学第二临床医学院，广东深圳 518020
  3.省部共建中医湿证国家重点实验室，广东广州 510120
  4.广东省中医急症研究重点实验室，广东广州 510120
  5.澳门科技大学医学院，澳门 999078
  6.广西国际壮医医院，广西南宁 530201
- 作者简介：
  
  LI Yan， E-mail： 091932@yzu.edu.cn
  CHENG Xiao， E-mail： chengxiaolucky@126.com
- 基金信息：
- DOI：10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.20240701.001
  CLC： R285.5
- Published：20 July 2024，
  
  Received：28 May 2024，
  
  Accepted：19 June 2024
- 稿件说明：
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王成毅,蔡粤芳,宁振求等.壮通饮通过Nrf2-SCL7A11/xCT-Gpx4通路调控铁死亡改善脑缺血再灌注损伤[J].中山大学学报（医学科学版）,2024,45(04):539-548.

WANG Chengyi,CAI Yuefang,NING Zhenqiu,et al.Zhuangtongyin Modulates Ferroptosis via the Nrf2-SCL7A11/xCT-Gpx4 Pathway to Improve Cerebral Ischemia-reperfusion Injury[J].Journal of Sun Yat-sen University(Medical Sciences),2024,45(04):539-548.
王成毅,蔡粤芳,宁振求等.壮通饮通过Nrf2-SCL7A11/xCT-Gpx4通路调控铁死亡改善脑缺血再灌注损伤[J].中山大学学报（医学科学版）,2024,45(04):539-548. DOI： 10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.20240701.001.

WANG Chengyi,CAI Yuefang,NING Zhenqiu,et al.Zhuangtongyin Modulates Ferroptosis via the Nrf2-SCL7A11/xCT-Gpx4 Pathway to Improve Cerebral Ischemia-reperfusion Injury[J].Journal of Sun Yat-sen University(Medical Sciences),2024,45(04):539-548. DOI： 10.13471/j.cnki.j.sun.yat-sen.univ（med.sci）.20240701.001.

摘要

目的

探究壮通饮通过Nrf2-SCL7A11/xCT-Gpx4通路调控铁死亡对大脑中动脉栓塞模型小鼠脑缺血再灌注损伤的保护作用及其机制。

方法

将C57BL/6J小鼠随机分为假手术组（Sham）、模型组（MCAO）、低剂量壮通饮组（ZTY-L+MCAO），高剂量壮通饮组（ZTY-H+MCAO），每组5只。其中MCAO组采用硅胶线栓法进行造模，小鼠大脑中动脉栓塞1h，拔出线栓再灌注72h后进行心脏灌注取脑，Sham组除了不插入线栓其余操作同MCAO组。采用Zea-Longa法对小鼠神经功能进行评分；采用TTC染色评估小鼠脑梗死体积；采用HE染色、尼氏染色评估脑损伤程度；采用普鲁士蓝染色评价三价铁离子蓄积水平；qPCR检测铁离子转运相关载体受体TfR1和DMT1、脂质过氧化相关基因ACSL4以及铁死亡标志物PTGS2 mRNA表达；ELISA试剂盒检测4-HNE的含量评估小鼠大脑脂质过氧化水平；Western-blot、免疫荧光检测小鼠脑组织中Nrf2、SCL7A11/xCT、Gpx4蛋白水平。

结果

①壮通饮改善了MCAO/R后小鼠神经功能（

＜0.05）和大脑梗死体积（

＜0.05）及缓解MCAO/R后大脑皮层细胞病理损伤。②壮通饮减弱了MCAO/R后小鼠大脑组织的三价铁离子蓄积状态；减少MCAO/R后小鼠大脑中与铁离子转运入细胞内相关载体TfR1和DMT1 mRNA的表达（

＜0.001）；下调MCAO/R后小鼠大脑脂质过氧化物4-HNE含量和组织脂质过氧化物酶ACSL4 mRNA表达（

＜0.001）；降低MCAO/R后小鼠大脑铁死亡标志物PTGS2 mRNA表达（

＜0.001）。③壮通饮增加MCAO/R后Nrf2入核表达（

＜0.001），以及增加xCT和Gpx4在神经元中的表达（

＜0.001）。

结论

壮通饮能够改善MCAO/R小鼠神经功能和脑梗死体积，缓解大脑皮层细胞病理损伤，影响Nrf2-SCL7A11/xCT-Gpx4通路关键信号分子表达，所以壮通饮改善小鼠MCAO/R损伤的作用机制可能是通过Nrf2-SCL7A11/xCT-Gpx4通路调节铁死亡水平达到的。

Abstract

Objective

To investigate the protective effect of Zhuangtongyin on the Middle Cerebral Artery Occlusion （MCAO） model by modulating ferroptosis through the Nrf2-SCL7A11/xCT-Gpx4 pathway and its underlying mechanism.

Methods

C57BL/6J mice were randomly divided into Sham operation group （Sham）， model group （MCAO）， low-dose Zhuangtongyin group （ZTY-L）， high-dose Zhuangtongyin group（ZTY-H）， with 5 mice in each group. The MCAO group was modelled by silica gel embolization， the middle cerebral artery of mice was embolized for 1h， then the silica gel was pulled out and reperfusion was performed after 72 h； and the other operations in the Sham group were the same as those in the MCAO group except that the thread plug was not inserted. The neural function of mice was evaluated by Zea-Longa method. TTC staining was used to evaluate the volume of cerebral infarction. The level of brain injury was evaluated by HE staining and Nissl staining. Prussian blue staining and the expression of iron transport-related carrier receptors TfR1 and DMT1 on mRNA level was detected by qPCR to evaluate the iron ion deposition level in mice brain. The expression of lipid peroxidation-related gene ACSL4 on mRNA level was detected by qPCR， and the content of 4-HNE was detected by ELISA kit to evaluate the lipid peroxidation level of mice brain. The expressions of ferroptosis marker PTGS2 mRNA level was detected by qPCR. The expressions of Nrf2， SCL7A11/xCT， Gpx4 in mice brain tissue were detected by Western-blot and immunofluorescence.

Results

Zhuangtongyin improved the nerve function of mice after MCAO （

＜0.05） and the cerebral infarction volume of mice （

＜0.05） and alleviate the pathological injury of cerebral cortex cells after MCAO operation. Zhuangtongyin attenuated the accumulation of trivalent iron ions in the brain tissue of mice following MCAO. Additionally， Zhuangtongyin downregulated the expression of TfR1 and DMT1 mRNA （

＜0.001）， a transporter associated wit

h cellular iron ion uptake， in the brains of post-MCAO mice. Furthermore， Zhuangtongyin reduced levels of lipid peroxidation product 4-HNE （

＜0.001） and suppressed ACSL4 mRNA expression in brain tissue post-MCAO （

＜0.001）. Besides， Zhuangtongyin downregulated the expression of PTGS2 mRNA （

＜0.001）， in the brains of post-MCAO mice. Zhuangtongyin increased the expression of nrf2 into the nucleus （

＜0.001）， and increased the expression of xCT and Gpx4 in neurons after MCAO （

＜0.001）.

Conclusion

Zhuangtongyin can enhance the nerve function and reduce cerebral infarction volume in MCAO/R mice， alleviate the pathological damage of cerebral cortex cells， and modulate the expression of key signaling molecules in the Nrf2-SCL7A11/xCT-Gpx4 pathway. Therefore， it is suggested that the mechanism by which Zhuangtongyin improves MCAO/R injury in mice may involve regulating ferroptosis through the Nrf2-SCL7A11/xCT-GPX4 pathway.

关键词

缺血性脑卒中铁死亡壮通饮Nrf2-SCL7A11/xCT-Gpx4通路机制研究

Keywords

ischemic strokeferroptosisZhuangtongyinNrf2-SCL7A11/xCT-Gpx4 pathwaymechanisms of action

references

思宇涛，尹琳，马春野，等．前后循环大血管闭塞型缺血性脑卒中血管成功再通后临床预后的比较［J］. 中华神经医学杂志，2023，2（10）：1016-1022.

Si YT， Yin L， Ma CY， et al. Comparison of clinical outcomes after successful revasc-ularization in anterior and posterior circulatory large vessel occlusive ischemic stroke［J］. Chin J Neuromed， 2023， 2（10）： 1016-1022.

陈孝男，杨爱琳，赵亚楠，等．缺血性脑中风的发病机制及其常用治疗中药研究进展［J］. 中国中药杂志，2019，44（3）：422-432.

Chen XN， Yang AL， Zhao YN， et al. Research progress on pathogenesis of ischemic cerebral apoplexy and its treatment with traditional Chinese medicine［J］. Chin J Chin Mater Med， 2019， 44（3）： 422-432.

Yan HF， Tuo QZ， Yin QZ， et al. The pathological role of ferroptosis in ischemia/repe-rfusion-related injury［J］. Zool Res， 2020， 41（3）： 220-230.

Dodson M， Castro-Portuguez R， Zhang DD. NRF2 plays a critical role in mitigating lipid peroxidation and ferroptosis［J］. Redox Biol， 2019， 23： 101107.

Xie Y， Kang R， Klionsky DJ， et al. GPX4 in cell death， autophagy， and disease［J］. Autophagy， 2023， 19（10）： 2621-2638.

安可，周学东. 铁死亡与骨关节病的研究新进展［J］. 四川大学学报（医学版）， 2023， 54（6）： 1294-1299.

An K， Zhou XD. Latest findings on ferroptosis and osteoarthropathy［J］. J Sichuan Univ （ Med Sci ）， 2023， 54（6）： 1294-1299.

Yang W， Wang Y， Zhang C， et al. Maresin1 protect against ferroptosis-induced liver injury through ROS inhibition and Nrf2/HO-1/GPX4 activation［J］. Front Pharmacol， 2022， 13： 865689.

Tong G， Wang X， Chen S， et al. Astragalus polysaccharide inhibits the development of urothelial carcinoma by activating AMPK signaling to induce BENC1-xCT complex formation［J］. Aging （Albany NY）， 2023， 15（18）： 9438-9452.

Bi R， Hu R， Jiang L， et al. Butyrate enhances erastin-induced ferroptosis of lung cancer cells via modulating the ATF3/SLC7A11 pathway［J］. Environ Toxicol， 2024， 39（2）： 529-538.

Li P， Chen JM， Ge SH， et al. Pentoxifylline protects against cerebral ischaemia-reperfusion injury through ferroptosis regulation via the Nrf2/SLC7A11/GPX4 signalling pathway［J］. Eur J Pharmacol， 2024， 967： 176402.

揭洁，陈莹，朱永娟，等．壮医水蛭疗法治疗麻邦的技术规范研究［J］. 中医临床研究，2023，15（19）：64-67.

Jie J， Chen Y， Zhu YJ， et al. Study on technical specification of Zhuangyi medical l-eech therapy for MaBang［J］. Clin J Chin Med， 2023， 15（19）： 64-67.

翟阳. 通龙路火路法治疗麻邦（中风）的理论探源［J］. 中国民间疗法，2023，31（3）：34-35； +63.

Zhai Y. The theoretical source of unclog Long Road and Fire Road treatment of Mabang （apoplexy）［J］. China’s Naturopathy， 2023， 31（3）： 34-35； +63.

李岩，覃启京，梁慧荟，等．壮通饮抑制NADPH氧化酶2对糖氧剥夺诱导星形胶质细胞损伤的保护作用［J］. 亚太传统医药，2020，16（9）：17-21.

Li Y， Tan QJ， Liang HH， et al. Zhuangtongyin inhibited the protective effect of NADPH oxidase-2 on astrocyte injury induced by glucose and oxygen deprivation［J］. Asia-Pacific Trad Med， 2020， 16（09）： 17-21.

赖思嘉，王凯华，杨鑫勇，等．壮通饮治疗缺血性卒中研究进展［J］. 亚太传统医药，2023，19（7）：195-199.

Lai SJ， Wang KH， Yang XY， et al. Research progress of Zhuangtongyin in the treatment of ischemic stroke［J］. Asia-Pacific Trad Med， 2023， 19（7）： 195-199.

李岩，王凯华，朱亮，等．壮通饮对缺氧/复氧诱导的HUVEC/U251细胞屏障损伤的影响［J］. 中国病理生理杂志，2022，38（1）：80-86.

Li Y， Wang KH， Zhu L， et al. Effect of Zhuangtonyin on HUVEC/U251 cell barrier damage induced by hypoxia/reoxygenation［J］. Chin J Pathophysiol， 2022， 38（1）： 80-86.

李岩，朱亮，赖思嘉，等．壮通饮调控昼夜节律对大鼠脑缺血损伤保护作用实验研究［J］.亚太传统医药，2022，18（10）：23-27.

Li Y， Zhu L， Lai SJ， et al. Experimental study on protective effect of Zhuangtonyin regulating circadian rhythm on cerebral ischemic injury in rats［J］. Asia-Pacific Trad Med， 2022， 18（10）： 23-27.

刘震乾，陈默，何磊，等．小鼠脑缺血再灌注模型稳定性研究［J］. 现代生物医学进展，2018，18（8）：1435-1440.

Liu ZQ， Chen M， He L， et al. Stability of cerebral ischemia-reperfusion model in mice［J］. Prog Modern Biomed， 2018， 18（8）： 1435-1440.

胡凯超，高岩，何佳琦，等．人参皂苷Rg1通过激活Nrf2/xCT/GPX4通路抑制神经元铁死亡改善缺血性脑卒中损伤［J］. 中国药理学通报，2023，39（10）：1905-1913.

Hu KC， Gao Y， He JQ， et al. Ginsenoside Rg1 improves ischemic stroke injury by inhibiting neuronal ferroptosis through activation of Nrf2/xCT/GPX4 pathway［J］. Chin Pharmacol Bull， 2023， 39（10）： 1905-1913.

Liu H， Zhang TA， Zhang WY， et al. Rhein attenuates cerebral ischemia-reperfusion injury via inhibition of ferroptosis through NRF2/SLC7A11/GPX4 pathway［J］. Exp Neurol， 2023， 369： 114541.

Yuan Y， Zhai Y， Chen J， et al. Kaempferol ameliorates oxygen-glucose deprivation/ r-eoxygenation-induced neuronal ferroptosis by activating Nrf2/SLC7A11/GPX4 axis［J］. Biomolecules， 2021 ， 11（7）： 923.

Liu X， Ren M， Zhang A， et al. Nrf2 attenuates oxidative stress to mediate the protective effect of ciprofol against cerebral ischemia-reperfusion injury［J］. Funct Integr Genomics， 2023， 23（4）： 345.

何俊荣，刘仔，陈锡培．布洛芬对缺血性脑中风大鼠的神经保护作用及对Nrf2/SLC7A11/GPX4信号通路的影响［J］. 实用医学杂志，2023，39（15）：1888-1892.

He JR， Liu Z， Chen XP， et al. Neuroprotective effect of ibuprofen on ischemic cerebral apoplectic rats and its effect on Nrf2/SLC7A11/GPX4 signaling pathway［J］. J Practical Med， 2023， 39（15）： 1888-1892.

杨鑫勇，王凯华，刘丹宁，等．壮通饮对脑缺血再灌注小鼠神经细胞损伤的影响及其作用机制［J］. 湖南中医药大学学报，2023，43（7）：1155-1164.

Yang XY， Wang KH， Liu DN， et al. Effect of Zhuangtongyin on nerve cell injury in cerebral ischemia-reperfusion mice and its mechanism［J］. J Tradi Chin Med， 2023， 43（7）： 1155-1164.

Sun X， Ou Z， Chen R， et al. Activation of the p62-Keap1-NRF2 pathway protects against ferroptosis in hepatocellular carcinoma cells［J］. Hepatology， 2016， 63（1）： 173-184.

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