| 安林,袁宇,韩翠玉,等.虫草素通过激活 Nrf2/HO-1/NQO1通路介导巨噬细胞极化进而缓解脑缺血再灌注损伤[J].安徽医药,2026,30(9):1755-1761. |
| 虫草素通过激活 Nrf2/HO-1/NQO1通路介导巨噬细胞极化进而缓解脑缺血再灌注损伤 |
| Cordycepin alleviates cerebral ischemia-reperfusion injury by mediating macrophage polarization through activation of the Nrf2/HO-1/NQO1 pathway |
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| DOI：10.3969/j.issn.1009-6469.2026.09.011 |
| 中文关键词: 虫草素 核转录因子红系 2相关因子 2 巨噬细胞 脑缺血再灌注损伤 血红素加氧酶 -1 |
| 英文关键词: Cordycepin Nuclear factor erythroid 2-related factor 2 Macrophages Cerebral ischemia-reperfusion injury Heme oxygenase-1 |
| 基金项目:河北省医学科学研究课题计划( 20231507) |
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| 中文摘要: |
| 目的探讨虫草素( Cor)通过核转录因子红系 2相关因子 2(Nrf2)/血红素加氧酶 -1(HO-1)/NAD(P)H:醌氧化还原酶 1(NQO1)通路对巨噬细胞极化介导的脑缺血再灌注损伤的作用。方法 2023年 8月至 2024年 5月，将 PC12神经细胞分为对照组、氧糖剥夺 /再灌注( OGD/R)组、 Cor干预组、依达拉奉组。以细胞计数试剂盒( CCK-8)法检测 PC12细胞活力，流式细胞术检测细胞凋亡。使用 Transwell共培养系统将各组 PC12细胞和 M1巨噬细胞共培养并将 PC12细胞分为 PC12+M1组、 OGD/R+M1组、 Cor(5 μmol/L)+M1组、 Cor(10 μmol/L)+M1组、 Cor(20 μmol/L)+M1组和依达拉奉 +M1组。以实时荧光定量逆转录聚合酶链反应( qRT-PCR)检测 PC12细胞中 CD86和 CD206的 mRNA水平，酶联免疫吸附分析( ELISA)检测 PC12细胞中白细胞介素 -4(IL-4)、肿瘤坏死因子 α(TNF-α)、白细胞介素 -1β(IL-1β)水平，蛋白质印迹法检测 PC12细胞中 Nrf2、HO-1和 NQO1的蛋白表达。另外采用随机数字表法将 50只无特定病原体( SPF)级雄性 C57BL/6小鼠分为假手术组、模型组、 Cor低剂量组、 Cor高剂量组、依达拉奉组，每组 10只。评价各组神经行为学评分，苏木精 -伊红(HE)染色检测各组脑组织病理损伤情况。 qRT-PCR检测各组小鼠脑组织中 CD86和 CD206的 mRNA水平， ELISA检测脑组织中 IL-4、TNF-α和 IL-1β水平，免疫组织化学染色检测脑组织中 Nrf2、HO-1和 NQO1的阳性表达。结果相较于对照组细胞活力(100.00±11.35)%和凋亡率(6.05±0.87)%，OGD/R组细胞活力( 33.59±5.26)%显著降低，凋亡率(38.46±2.79)%显著升高，而 Cor或依达拉奉处理显著促进细胞活力，抑制细胞凋亡(均 P<0.05)。与 PC12+M1组相比， OGD/R+M1组细胞的 CD86、TNF-α和 IL-1β水平升高， CD206和 IL-4水平降低(均 P<0.05)。而相较于 OGD/R+M1组， Cor或依达拉奉干预则抑制 CD86、TNF-α和 IL-1β水平，促进 CD206和 IL-4水平(均 P<0.05)。相对于 PC12+M1组， OGD/R+M1组的 Nrf2、HO-1和 NQO1蛋白表达降低，而相较于 OGD/R+M1组， Cor+M1组和依达拉奉 +M1组的 Nrf2、HO-1和 NQO1蛋白表达升高(均 P<0.05)。动物实验结果显示，与模型组比较，低剂量和高剂量 Cor均能够显著改善大脑中动脉闭塞 /再灌注( MCAO/R)小鼠的神经损伤和炎症反应，促进 M2巨噬细胞相关因子的表达，激活 Nrf2、HO-1和 NQO1的阳性表达且高剂量 Cor效果更显著(均 P<0.05)。结论 Cor通过激活 Nrf2/HO-1/NQO1通路介导巨噬细胞极化进而缓解脑缺血再灌注损伤。 |
| 英文摘要: |
| Objective To explore the effect of cordycepin (Cor) on macrophage polarization-mediated cerebral ischemia-reperfusion injury through the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1)/NAD(P)H: quinone oxidoreductase 1 (NQO1) pathway. Methods From August 2023 to May 2024, PC12 cells were divided into the control group, oxygen-glucose depriva.tion/reoxygenation (OGD/R) group, Cor intervention group, and edaravone group. Cell viability of PC12 cells was detected by cell count.ing kit-8 (CCK-8), and cell apoptosis was detected by flow cytometry. PC12 cells of each group were co-cultured with M1 macrophages using a Transwell co-culture system, and PC12 cells were divided into the PC12+M1 group, OGD/R+M1 group, Cor (5 μmol/L)+M1group, Cor (10 μmol/L)+M1 group, Cor (20 μmol/L)+M1 group, and edaravone+M1 group. The mRNA levels of CD86 and CD206 inPC12 cells were detected by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). The levels of interleu. kin-4 (IL-4), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) in PC12 cells were detected by enzyme-linked immunosor. bent assay (ELISA). The protein expressions of Nrf2, HO-1, and NQO1 in PC12 cells were detected by Western blotting. Fifty SPF-grade male C57BL/6 mice were divided into the sham operation group, model group, low-dose Cor group, high-dose Cor group, and edar.avone group using the random number table method, with 10 mice in each group. The neurobehavioral scores of each group were evalu.ated, and the pathological damage of brain tissue in each group was detected by hematoxylin-eosin (HE) staining. The mRNA levels of CD86 and CD206 in the brain tissue of mice in each group were detected by qRT-PCR. The levels of IL-4, TNF-α, and IL-1β in brain tissue were detected by ELISA. The positive expressions of Nrf2, HO-1, and NQO1 in brain tissue were detected by immunohistochemi. cal staining.Results Compared with the control group with cell viability (100.00±11.35)% and apoptosis rate (6.05±0.87)%, the OGD/R group exhibited significantly reduced cell viability (33.59±5.26)% and increased apoptosis rate (38.46±2.79)% . In contrast, treat.ment with Cor or edaravone significantly enhanced cell vitality and inhibited apoptosis (all P<0.05). Compared with the PC12+M1 group, the OGD/R+M1 group exhibited increased levels of CD86, TNF-α, and IL-1β, along with decreased levels of CD206 and IL-4 (all P<0.05). In contrast to the OGD/R+M1 group, intervention with Cor or edaravone inhibited the levels of CD86, TNF-α, and IL-1β, while promoting the levels of CD206 and IL-4 (all P<0.05). Compared with the PC12+M1 group, the OGD/R+M1 group showed de. creased protein expression of Nrf2, HO-1, and NQO1, while compared with the OGD/R+M1 group, the Cor+M1 group and the edara.vone+M1 group showed increased protein expression of Nrf2, HO-1, and NQO1 (all P<0.05). Animal experiment results indicated that, compared with the model group, both low-dose and high-dose Cor significantly improved nerve injury and inflammatory response inmice with middle cerebral artery occlusion/reperfusion (MCAO/R), promoted the expression of M2 macrophage-related factors, and acti. vated the positive expression of Nrf2, HO-1, and NQO1, with high-dose Cor exhibiting more pronounced effects (all P<0.05).Conclu. sion Cor alleviates cerebral ischemia-reperfusion injury by activating the Nrf2/HO-1/NQO1 pathway to mediate macrophage polariza. tion. |
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