中国组织工程研究

中国组织工程研究 ›› 2024, Vol. 28 ›› Issue (7): 1063-1069.doi: 10.12307/2024.119

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

羟基红花黄色素A干预脑缺血再灌注损伤后星形胶质细胞脂质运载蛋白2的表达

刘可心1，宋丽娟1，2，3，吴艺舸1，韩光远1，苗珠月1，魏汝恒1，肖保国4，马存根1，黄建军1，3   

  1. 1山西中医药大学国家中医药管理局多发性硬化益气活血重点研究室/神经生物学研究中心，山西省晋中市   030619；2山西医科大学细胞生理学省部共建教育部重点实验室，山西省太原市   030001；3国药同煤集团总医院神经外科，山西省大同市   037003；4复旦大学神经病学研究所、脑科学研究院和医学神经生物学国家重点实验室，上海市   200025
  • 收稿日期:2022-12-30 接受日期:2023-03-14 出版日期:2024-03-08 发布日期:2023-07-17
  • 通讯作者: 黄建军，硕士，主任医师，山西中医药大学国家中医药管理局多发性硬化益气活血重点研究室/神经生物学研究中心，山西省晋中市 030619；国药同煤集团总医院神经外科，山西省大同市 037003 宋丽娟，博士，副教授，山西中医药大学国家中医药管理局多发性硬化益气活血重点研究室/神经生物学研究中心，山西省晋中市 030619；山西医科大学细胞生理学省部共建教育部重点实验室，山西省太原市 030001；国药同煤集团总医院神经外科，山西省大同市 037003
  • 作者简介:刘可心，女，1994年生，河北省保定市人，汉族，2023年山西中医药大学毕业，硕士，主要从事中医药防治缺血性脑血管疾病的研究。 宋丽娟，女，1985年生，山西省太原市人，汉族，博士，副教授，主要从事中医药防治缺血性脑血管疾病的研究。
  • 基金资助:
    国家自然科学基金(82004028)，项目负责人：宋丽娟；国药同煤总医院横向课题(202209SY01)，项目负责人：宋丽娟；中国博士后科学基金面上资助项目(2020M680912)，项目负责人：宋丽娟；2021年山西中医药大学附属医院国家区域中医医疗中心心血管专项基金项目(XGZX202115)，项目负责人：宋丽娟；山西中医药大学学科建设经费，项目负责人：马存根；山西省卫健委医学科技领军团队(2020TD05)，项目负责人：马存根；山西中医药大学2021年科技创新能力培育计划“青年科学家培育专题”项目(2021PY-QN-09)，项目负责人：宋丽娟；山西中医药大学2022年度科技创新团队(2022TD2010)，项目负责人：宋丽娟；2022年度山西省科技创新人才团队(202204051001028)，项目负责人：宋丽娟

Hydroxysafflor yellow A intervenes astrocyte lipocalin 2 expression after cerebral ischemia/reperfusion injury

Liu Kexin1, Song Lijuan1, 2 , 3, Wu Yige1, Han Guangyuan1, Miao Zhuyue1, Wei Ruheng1, Xiao Baoguo4, Ma Cungen1, Huang Jianjun1, 3   

  1. 1Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Chinese Medicine/Research Center of Neurobiology, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China; 2Key Laboratory of Cell Physiology Jointly Established by Province and Ministry of Education, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China; 3Department of Neurosurgery, Sinopharm Tongmei General Hospital, Datong 037003, Shanxi Province, China; 4Institute of Neurology, Institutes of Brain Science and State Key Laboratory of Medical Neurobiology, Fudan University, Shanghai 200025, China
  • Received:2022-12-30 Accepted:2023-03-14 Online:2024-03-08 Published:2023-07-17
  • Contact: Huang Jianjun, Master, Chief physician, Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Chinese Medicine/Research Center of Neurobiology, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China; Department of Neurosurgery, Sinopharm Tongmei General Hospital, Datong 037003, Shanxi Province, China Song Lijuan, MD, Associate professor, Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Chinese Medicine/Research Center of Neurobiology, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China; Key Laboratory of Cell Physiology Jointly Established by Province and Ministry of Education, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China; Department of Neurosurgery, Sinopharm Tongmei General Hospital, Datong 037003, Shanxi Province, China
  • About author:Liu Kexin, Master, Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Chinese Medicine/Research Center of Neurobiology, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China Song Lijuan, MD, Associate professor, Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Chinese Medicine/Research Center of Neurobiology, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China; Key Laboratory of Cell Physiology Jointly Established by Province and Ministry of Education, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China; Department of Neurosurgery, Sinopharm Tongmei General Hospital, Datong 037003, Shanxi Province, China
  • Supported by:
    National Natural Science Foundation of China, No. 82004028 (to SLJ); Horizontal Project of Sinopharm Tongmei General Hospital, No. 202209SY01 (to SLJ); China Postdoctoral Science Foundation, No. 2020M680912 (to SLJ); Cardiovascular Special Fund Project of National Regional Traditional Chinese Medicine Medical Center of Affiliated Hospital of Shanxi University of Chinese Medicine in 2021, No. XGZX202115 (to SLJ); Discipline Construction Fund of Shanxi University of Chinese Medicine (to MCG); Leading Team of Medical Science and Technology of Shanxi Province, No. 2020TD05 (to MCG); Young Scientists Cultivation Project of Shanxi University of Chinese Medicine in 2021, No. 2021PY-QN-09 (to SLJ); The 2022 Scientific and Technological Innovation Team of Shanxi University of Chinese Medicine, No. 2022TD2010 (to SLJ); 2022 Shanxi Provincial Science and Technology Innovation Talent Team, No. 202204051001028 (to SLJ)

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摘要：


文题释义：

脑缺血：由于脑血管狭窄或闭塞引起相应动脉支配供血的脑组织发生暂时性功能障碍。
脂质运载蛋白2：又称中性粒细胞明胶酶相关脂质蛋白(NGAL)，也被称为24p3，参与多种生物反应，其作为反应性星形胶质细胞的重要标志物，参与了脑卒中后继发性损伤的病理生理过程。


背景：缺血性脑卒中严重威胁人类健康，缺血缺氧后星形胶质细胞大量表达脂质运载蛋白2加重脑损伤，但其具体机制并不清楚。羟基红花黄色素A具有抗缺血、抗氧化、抗血栓及抗炎等作用，其是否影响脑缺血缺氧后星形胶质细胞表达脂质运载蛋白2，目前尚不清楚。

目的：探究羟基红花黄色素A对脑缺血再灌注损伤后星形胶质细胞中脂质运载蛋白2表达的影响及机制。
方法：①将30只成年SD大鼠随机分成3组：假手术组、大脑中动脉闭塞再灌注组、羟基红花黄色素A组，后2组建立大脑中动脉闭塞再灌注模型，羟基红花黄色素A组在再灌注后以12 mg/kg的剂量腹腔注射羟基红花黄色素A。采用Longa评分法评估神经功能缺损程度，采用TTC染色法测定脑梗死体积，Western blot和免疫荧光检测JAK2/STAT3通路及脂质运载蛋白2的表达，ELISA法检测白细胞介素1β、白细胞介素6、肿瘤坏死因子α水平。②将星形胶质细胞分为4组：正常组、糖氧剥夺复糖氧组、羟基红花黄色素A组、AG490组，后3组建立糖氧剥夺复糖氧模型，在糖氧剥夺期间分别用75 μmol/L羟基红花黄色素A、10 μmol/L酪氨酸磷酸化抑制剂AG490处理星形胶质细胞8 h，进一步验证羟基红花黄色素A对脂质运载蛋白2的作用机制。

结果与结论：①与假手术组相比，大脑中动脉闭塞再灌注组大鼠脑梗死体积明显增加，并伴有神经功能损伤加重(P < 0.01)，羟基红花黄色素A治疗可以减小脑梗死体积，改善神经功能(P < 0.01)；②大脑中动脉闭塞再灌注组p-JAK2、p-STAT3和脂质运载蛋白2的表达高于假手术组(P < 0.01)，羟基红花黄色素A治疗后抑制了p-JAK2、p-STAT3和脂质运载蛋白2的表达(P < 0.01)；③大脑中动脉闭塞再灌注组炎性因子白细胞介素1β、白细胞介素6、肿瘤坏死因子α水平高于假手术组(P < 0.01)，羟基红花黄色素A治疗后抑制了白细胞介素1β、白细胞介素6、肿瘤坏死因子α表达(P < 0.01)；④体外实验糖氧剥夺复糖氧组p-JAK2、p-STAT3、脂质运载蛋白2的表达高于正常组(P < 0.01)，加入AG490后JAK2、STAT3磷酸化降低，脂质运载蛋白2表达被抑制(P < 0.01)。结果表明，羟基红花黄色素A可能通过调控JAK2/STAT3信号通路抑制缺血缺氧后星形胶质细胞中脂质运载蛋白2的表达，从而减轻脑损伤。

https://orcid.org/0000-0003-2945-5669 (宋丽娟) 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 脑缺血, 星形胶质细胞, 羟基红花黄色素A, 脂质运载蛋白2, JAK2, STAT3

Abstract: BACKGROUND: Ischemic stroke is a serious threat to human health. After ischemia and hypoxia, astrocyte expresses lipocalin-2 in large amounts to aggravate brain injury, but the specific mechanism is not clear. Hydroxysafflor yellow A has anti-ischemia, anti-oxidation, anti-thrombosis and anti-inflammatory effects. However, whether hydroxysafflor yellow A affects the expression of lipocalin-2 in astrocytes after cerebral ischemia and hypoxia and its mechanism are not clear. 
OBJECTIVE: To investigate the effect and mechanism of hydroxysafflor yellow A on the expression of lipocalin-2 in astrocytes after cerebral ischemia and reperfusion.
METHODS: (1) Thirty adult SD rats were randomly divided into three groups: sham operation group, middle cerebral artery occlusion and reperfusion group, and hydroxysafflor yellow A group. The middle cerebral artery occlusion and reperfusion model was established in the latter two groups, and hydroxysafflor yellow A group was intraperitoneally injected with 12 mg/kg hydroxysafflor yellow A after reperfusion. Longa score was used to evaluate the degree of neurological impairment. Infarct volume was determined by TTC staining. JAK2/STAT3 pathway and lipocalin-2 expression were detected by western blot assay and immunofluorescence. Levels of interleukin 1β, interleukin 6 and tumor necrosis factor α were detected by ELISA. (2) Astrocytes were divided into four groups: Normal group, glucose-oxygen deprivation group, hydroxysafflor yellow A group and AG490 group. In the latter three groups, glucose-oxygen deprivation and glucose-oxygen recovery models were established. Astrocytes were treated with 75 μmol/L hydroxysafflor yellow A and 10 μmol/L tyrosine phosphorylation inhibitor AG490 for 8 hours during glucose-oxygen deprivation, respectively. The mechanism of hydroxysafflor yellow A on lipocalin-2 was further verified.  
RESULTS AND CONCLUSION: (1) Compared with the sham operation group, cerebral infarction was significantly increased in the middle cerebral artery occlusion and reperfusion group, accompanied by aggravated neurological impairment (P < 0.01). Hydroxysafflor yellow A treatment could reduce cerebral infarction volume and improve neurological function (P < 0.01). (2) The expressions of p-JAK2, p-STAT3 and lipocalin-2 in the middle cerebral artery occlusion and reperfusion group were higher than those in the sham operation group (P < 0.01). Hydroxysafflor yellow A treatment reduced the expressions of JAK2, STAT3 and lipocalin-2 (P < 0.01). (3) The expression levels of interleukin 1β, interleukin-6 and tumor necrosis factor α in the middle cerebral artery occlusion and reperfusion group were higher than those in the sham operation group (P < 0.01). Hydroxysafflor yellow A inhibited the expressions of interleukin 1β, interleukin-6 and tumor necrosis factor α (P < 0.01). (4) In vitro, the expressions of p-JAK2, p-STAT3 and lipocalin-2 in the glucose-oxygen deprivation group were significantly higher than those in the normal group (P < 0.01). After adding AG490, the phosphorylation of JAK2 and STAT3 decreased, and the expression of lipocalin-2 was inhibited (P < 0.01). The results suggest that hydroxysafflor yellow A may inhibit the expression of lipocalin-2 in astrocytes after ischemia and hypoxia by regulating the JAK2/STAT3 signaling pathway, thereby reducing brain injury.

Key words: cerebral ischemia, astrocyte, hydroxysafflor yellow A, lipocalin-2, JAK2, STAT3

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