Article(id=1292135558730510974, tenantId=1146029695717560320, journalId=1246415772164075586, issueId=1292135425250976333, articleNumber=null, orderNo=null, doi=10.13699/j.cnki.1001-6821.2026.10.006, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1762704000000, receivedDateStr=2025-11-10, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1785998723918, onlineDateStr=2026-08-06, pubDate=1779897600000, pubDateStr=2026-05-28, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1785998723918, onlineIssueDateStr=2026-08-06, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1785998723918, creator=13701087609, updateTime=1785998723918, updator=13701087609, issue=Issue{id=1292135425250976333, tenantId=1146029695717560320, journalId=1246415772164075586, year='2026', volume='42', issue='10', pageStart='1351', pageEnd='1500', issueExtLink='null', onlineDate='null', pubDate='1779897600000', pubDateStr='2026-05-28', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1785998692095, creator='13701087609', updateTime=1786014373634, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1292201198489202995, tenantId=1146029695717560320, journalId=1246415772164075586, issueId=1292135425250976333, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1292201198489202996, tenantId=1146029695717560320, journalId=1246415772164075586, issueId=1292135425250976333, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=1384, endPage=1390, ext={EN=ArticleExt(id=1292135560345318015, articleId=1292135558730510974, tenantId=1146029695717560320, journalId=1246415772164075586, language=EN, title=Research of wogonin on neuronal damage induced by hypoxia/reoxygenation based on the CCL2-CCR2 pathway, columnId=1246531407326105792, journalTitle=Chinese Journal of Clinical Pharmacology, columnName=Clinical and Basic Bridging Research, runingTitle=null, highlight=null, articleAbstract=
Objective

To investigate the effect of wogonin on hypoxia/reoxygenation (H/R)-induced neuronal injury based on the C-C chemokine ligand 2-C-C chemokine receptor 2 (CCL2-CCR2) pathway.

Methods

PC12 cells were cultured and divided into five groups, control group (normal culture), model group (H/R injury), experimental group (H/R+50 μmol·L-1 wogonin pretreatment for 1 h), antagonist group (H/R+10 μmol·L-1 CCR2 antagonist RS102895 pretreatment for 1 h), and agonist group (H/R+50 μmol·L-1 wogonin+20 μmol·L-1 CCL2 recombinant protein pretreatment for 1 h). Cell viability and cytotoxicity were assessed using the cell counting kit-8 (CCK-8) and lactate dehydrogenase (LDH) assay kits. Apoptosis was detected by Annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) double staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining. Oxidative stress and inflammatory responses were evaluated by enzyme-linked immunosorbent assay (ELISA) and quantitative real-time polymerase chain reaction (RT-qPCR). Protein expression levels related to the CCL2-CCR2 signaling pathway were measured by Western blot.

Results

The viability of PC12 cells in control group, model group, experimental group, antagonist group and agonist group were 0.89±0.06, 0.32±0.04, 0.69±0.05, 0.65±0.07 and 0.46±0.04, respectively; the SOD activities were (45.11±3.02), (10.05±1.05), (33.62±3.15), (30.15±2.87) and (21.05±2.17) U·mL-1, respectively; the Annexin V FITC/PI double staining apoptosis rates were (3.21±0.68)%, (29.65±1.59)%, (8.99±0.82)%, (9.87±0.85)% and (18.65±1.06)%, respectively; the TUNEL positive rates were (5.66±0.89)%, (33.65±2.42)%, (13.25±1.05)%, (11.66±1.87)% and (24.06±1.16)%, respectively; the relative expression levels of tumor necrosis factor-alpha(TNF-α) mRNA were 1.02±0.15, 3.05±0.32, 1.59±0.16, 2.98±0.24 and 2.66±0.20, respectively; the relative expression levels of interleukin-6 (IL-6) mRNA were 1.05±0.12, 4.54±0.39, 1.87±0.15, 1.75±0.14 and 3.08±0.32, respectively; the relative expression levels of CCL2 mRNA were 1.01±0.07, 3.99±0.32, 1.64±0.15, 3.90±0.33 and 2.84±0.23, respectively; the relative expression levels of CCR2 mRNA were 1.03±0.10, 5.21±0.51, 2.03±0.26, 2.11±0.20 and 3.85±0.32, respectively; the relative expression levels of phosphorylation (p)p65/p65 were 0.12±0.02, 0.75±0.06, 0.24±0.03, 0.22±0.01 and 0.52±0.05, respectively; the leakage rates of LDH were (18.32±1.15)%, (58.64±5.16)%, (29.02±2.16)%, (26.98±3.02)% and (40.24±3.54)%, respectively; the malondialdehyde (MDA) contents were (50.16±5.24), (169.65±10.16), (91.32±7.12), (86.21±5.16) and (134.16±11.87) nmol·mL-1, respectively. The differences of above indicators between model group and control group, between experimental group and model group, and between the antagonist, agonist group and experimental group were all statistically significant (all P<0.05).

Conclusion

Wogonin exerts a protective effect against H/R-induced neuronal injury by inhibiting the CCL2-CCR2 signaling pathway.

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目的

基于C-C趋化因子配体2-C-C趋化因子受体2(CCL2-CCR2)通路探讨汉黄芩素对缺氧复氧(H/R)诱导神经元损伤的作用。

方法

体外培养PC12细胞,依次分为对照组(常规培养)、模型组(H/R处理)、实验组(H/R+50 μmol·L-1汉黄芩素预处理1 h)、拮抗剂组(H/R+10 μmol·L-1 CCR2拮抗剂RS102895预处理1 h)和激动剂组(H/R+50 μmol·L-1汉黄芩素+20 μmol·L-1 CCL2重组蛋白预处理1 h)。用细胞计数试剂盒-8(CCK-8)和乳酸脱氢酶(LDH)试剂盒法检测细胞活力和毒性;用膜联蛋白V-异硫氰酸荧光素/碘化丙啶(Annexin V-FITC/PI)双染法和脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TUNEL)染色法检测细胞凋亡;用酶联免疫吸附试验(ELISA)法和实时荧光定量聚合酶链反应(RT-qPCR)法检测氧化应激和炎症反应;用蛋白质印迹法检测CCL2-CCR2信号通路相关蛋白表达。

结果

对照组、模型组、实验组、拮抗剂组和激动剂组的PC12细胞活力分别为0.89±0.06、0.32±0.04、0.69±0.05、0.65±0.07和0.46±0.04,SOD活性分别为(45.11±3.02)、(10.05±1.05)、(33.62±3.15)、(30.15±2.87)和(21.05±2.17)U·mL-1,Annexin VFITC/PI双染凋亡率分别为(3.21±0.68)%、(29.65±1.59)%、(8.99±0.82)%、(9.87±0.85)%和(18.65±1.06),TUNEL阳性率分别为(5.66±0.89)%、(33.65±2.42)%、(13.25±1.05)%、(11.66±1.87)%和(24.06±1.16)%,肿瘤坏死因子-α(TNFα)mRNA相对表达水平分别为1.02±0.15、3.05±0.32、1.59±0.16、2.98±0.24和2.66±0.20,白细胞介素-6(IL-6)mRNA相对表达水平分别为1.05±0.12、4.54±0.39、1.87±0.15、1.75±0.14和3.08±0.32,CCL2 mRNA相对表达水平分别为1.01±0.07、3.99±0.32、1.64±0.15、3.90±0.33和2.84±0.23,CCR2 mRNA相对表达水平分别为1.03±0.10、5.21±0.51、2.03±0.26、2.11±0.20和3.85±0.32,磷酸化(pp65/p65相对表达水平分别为0.12±0.02、0.75±0.06、0.24±0.03、0.22±0.01和0.52±0.05,LDH漏出率分别为(18.32±1.15)%、(58.64±5.16)%、(29.02±2.16)%、(26.98±3.02)%和(40.24±3.54)%,丙二醛(MDA)含量分别为(50.16±5.24)、(169.65±10.16)、(91.32±7.12)、(86.21±5.16)和(134.16±11.87)nmol·mL-1。模型组与对照组比较、实验组与模型组对比、拮抗剂组和激动剂组与实验组对比,上述指标在统计学上差异均有统计学意义(均P<0.05)。

结论

汉黄芩素通过抑制CCL2-CCR2信号通路对H/R诱导的神经元损伤具有保护作用。

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李启超(1990-)男,主治医师,主要从事脑卒中及卒中后康复评估方面的临床工作和研究

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基于CCL2-CCR2通路探讨汉黄芩素对缺氧复氧诱导神经元损伤的研究
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李启超 1a , 刘媛媛 1b , 黄占 1a
中国临床药理学杂志 | 临床与基础桥接研究 2026,42(10): 1384-1390
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中国临床药理学杂志 |临床与基础桥接研究 2026 , 42 (10) : 1384 -1390
基于CCL2-CCR2通路探讨汉黄芩素对缺氧复氧诱导神经元损伤的研究
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李启超1a, 刘媛媛1b, 黄占1a
作者信息
  • a.滕州市中心人民医院,康复医学科,山东 滕州 277500
  • b.滕州市中心人民医院,神经内科,山东 滕州 277500
通讯作者:
黄占,主治医师 MP: 15564790919 E-mail:
作者简介:

李启超(1990-)男,主治医师,主要从事脑卒中及卒中后康复评估方面的临床工作和研究

Research of wogonin on neuronal damage induced by hypoxia/reoxygenation based on the CCL2-CCR2 pathway
Qi-chao LI1a, Yuan-yuan LIU1b, Zhan HUANG1a
Affiliations
  • a.Department of Rehabilitation Medicine, Tengzhou Central People’s Hospital, Tengzhou 277500, Shandong Province, China
  • b.Department of Neurology, Tengzhou Central People’s Hospital, Tengzhou 277500, Shandong Province, China
出版时间: 2026-05-28 doi: 10.13699/j.cnki.1001-6821.2026.10.006
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目的

基于C-C趋化因子配体2-C-C趋化因子受体2(CCL2-CCR2)通路探讨汉黄芩素对缺氧复氧(H/R)诱导神经元损伤的作用。

方法

体外培养PC12细胞,依次分为对照组(常规培养)、模型组(H/R处理)、实验组(H/R+50 μmol·L-1汉黄芩素预处理1 h)、拮抗剂组(H/R+10 μmol·L-1 CCR2拮抗剂RS102895预处理1 h)和激动剂组(H/R+50 μmol·L-1汉黄芩素+20 μmol·L-1 CCL2重组蛋白预处理1 h)。用细胞计数试剂盒-8(CCK-8)和乳酸脱氢酶(LDH)试剂盒法检测细胞活力和毒性;用膜联蛋白V-异硫氰酸荧光素/碘化丙啶(Annexin V-FITC/PI)双染法和脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TUNEL)染色法检测细胞凋亡;用酶联免疫吸附试验(ELISA)法和实时荧光定量聚合酶链反应(RT-qPCR)法检测氧化应激和炎症反应;用蛋白质印迹法检测CCL2-CCR2信号通路相关蛋白表达。

结果

对照组、模型组、实验组、拮抗剂组和激动剂组的PC12细胞活力分别为0.89±0.06、0.32±0.04、0.69±0.05、0.65±0.07和0.46±0.04,SOD活性分别为(45.11±3.02)、(10.05±1.05)、(33.62±3.15)、(30.15±2.87)和(21.05±2.17)U·mL-1,Annexin VFITC/PI双染凋亡率分别为(3.21±0.68)%、(29.65±1.59)%、(8.99±0.82)%、(9.87±0.85)%和(18.65±1.06),TUNEL阳性率分别为(5.66±0.89)%、(33.65±2.42)%、(13.25±1.05)%、(11.66±1.87)%和(24.06±1.16)%,肿瘤坏死因子-α(TNFα)mRNA相对表达水平分别为1.02±0.15、3.05±0.32、1.59±0.16、2.98±0.24和2.66±0.20,白细胞介素-6(IL-6)mRNA相对表达水平分别为1.05±0.12、4.54±0.39、1.87±0.15、1.75±0.14和3.08±0.32,CCL2 mRNA相对表达水平分别为1.01±0.07、3.99±0.32、1.64±0.15、3.90±0.33和2.84±0.23,CCR2 mRNA相对表达水平分别为1.03±0.10、5.21±0.51、2.03±0.26、2.11±0.20和3.85±0.32,磷酸化(pp65/p65相对表达水平分别为0.12±0.02、0.75±0.06、0.24±0.03、0.22±0.01和0.52±0.05,LDH漏出率分别为(18.32±1.15)%、(58.64±5.16)%、(29.02±2.16)%、(26.98±3.02)%和(40.24±3.54)%,丙二醛(MDA)含量分别为(50.16±5.24)、(169.65±10.16)、(91.32±7.12)、(86.21±5.16)和(134.16±11.87)nmol·mL-1。模型组与对照组比较、实验组与模型组对比、拮抗剂组和激动剂组与实验组对比,上述指标在统计学上差异均有统计学意义(均P<0.05)。

结论

汉黄芩素通过抑制CCL2-CCR2信号通路对H/R诱导的神经元损伤具有保护作用。

汉黄芩素  /  缺氧复氧  /  神经元损伤  /  C-C趋化因子配体2- C-C趋化因子受体2通路
Objective

To investigate the effect of wogonin on hypoxia/reoxygenation (H/R)-induced neuronal injury based on the C-C chemokine ligand 2-C-C chemokine receptor 2 (CCL2-CCR2) pathway.

Methods

PC12 cells were cultured and divided into five groups, control group (normal culture), model group (H/R injury), experimental group (H/R+50 μmol·L-1 wogonin pretreatment for 1 h), antagonist group (H/R+10 μmol·L-1 CCR2 antagonist RS102895 pretreatment for 1 h), and agonist group (H/R+50 μmol·L-1 wogonin+20 μmol·L-1 CCL2 recombinant protein pretreatment for 1 h). Cell viability and cytotoxicity were assessed using the cell counting kit-8 (CCK-8) and lactate dehydrogenase (LDH) assay kits. Apoptosis was detected by Annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) double staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining. Oxidative stress and inflammatory responses were evaluated by enzyme-linked immunosorbent assay (ELISA) and quantitative real-time polymerase chain reaction (RT-qPCR). Protein expression levels related to the CCL2-CCR2 signaling pathway were measured by Western blot.

Results

The viability of PC12 cells in control group, model group, experimental group, antagonist group and agonist group were 0.89±0.06, 0.32±0.04, 0.69±0.05, 0.65±0.07 and 0.46±0.04, respectively; the SOD activities were (45.11±3.02), (10.05±1.05), (33.62±3.15), (30.15±2.87) and (21.05±2.17) U·mL-1, respectively; the Annexin V FITC/PI double staining apoptosis rates were (3.21±0.68)%, (29.65±1.59)%, (8.99±0.82)%, (9.87±0.85)% and (18.65±1.06)%, respectively; the TUNEL positive rates were (5.66±0.89)%, (33.65±2.42)%, (13.25±1.05)%, (11.66±1.87)% and (24.06±1.16)%, respectively; the relative expression levels of tumor necrosis factor-alpha(TNF-α) mRNA were 1.02±0.15, 3.05±0.32, 1.59±0.16, 2.98±0.24 and 2.66±0.20, respectively; the relative expression levels of interleukin-6 (IL-6) mRNA were 1.05±0.12, 4.54±0.39, 1.87±0.15, 1.75±0.14 and 3.08±0.32, respectively; the relative expression levels of CCL2 mRNA were 1.01±0.07, 3.99±0.32, 1.64±0.15, 3.90±0.33 and 2.84±0.23, respectively; the relative expression levels of CCR2 mRNA were 1.03±0.10, 5.21±0.51, 2.03±0.26, 2.11±0.20 and 3.85±0.32, respectively; the relative expression levels of phosphorylation (p)p65/p65 were 0.12±0.02, 0.75±0.06, 0.24±0.03, 0.22±0.01 and 0.52±0.05, respectively; the leakage rates of LDH were (18.32±1.15)%, (58.64±5.16)%, (29.02±2.16)%, (26.98±3.02)% and (40.24±3.54)%, respectively; the malondialdehyde (MDA) contents were (50.16±5.24), (169.65±10.16), (91.32±7.12), (86.21±5.16) and (134.16±11.87) nmol·mL-1, respectively. The differences of above indicators between model group and control group, between experimental group and model group, and between the antagonist, agonist group and experimental group were all statistically significant (all P<0.05).

Conclusion

Wogonin exerts a protective effect against H/R-induced neuronal injury by inhibiting the CCL2-CCR2 signaling pathway.

wogonin  /  hypoxia/reoxygenation  /  neuronal damage  /  C-C chemokine ligand 2-C-C chemokine receptor 2 pathway
李启超, 刘媛媛, 黄占. 基于CCL2-CCR2通路探讨汉黄芩素对缺氧复氧诱导神经元损伤的研究. 中国临床药理学杂志, 2026 , 42 (10) : 1384 -1390 . DOI: 10.13699/j.cnki.1001-6821.2026.10.006
Qi-chao LI, Yuan-yuan LIU, Zhan HUANG. Research of wogonin on neuronal damage induced by hypoxia/reoxygenation based on the CCL2-CCR2 pathway[J]. Chinese Journal of Clinical Pharmacology, 2026 , 42 (10) : 1384 -1390 . DOI: 10.13699/j.cnki.1001-6821.2026.10.006
  • 徐州医科大学附属医院发展基金资助项目(XYFY202214)
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doi: 10.13699/j.cnki.1001-6821.2026.10.006
  • 接收时间:2025-11-10
  • 首发时间:2026-08-06
  • 出版时间:2026-05-28
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  • 收稿日期:2025-11-10
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徐州医科大学附属医院发展基金资助项目(XYFY202214)
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    a.滕州市中心人民医院,康复医学科,山东 滕州 277500
    b.滕州市中心人民医院,神经内科,山东 滕州 277500

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黄占,主治医师 MP: 15564790919 E-mail:
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