槲皮素促进应激颗粒G3BP1解聚改善HIV-1 gp120诱导的星形胶质细胞神经毒性

doi:10.12122/j.issn.1673-4254.2025.02.11

摘要/Abstract

摘要：

目的探究槲皮素（QC）在HIV-1 gp120诱导的星形胶质细胞神经毒性中的抑制作用及机制。方法分离原代星形胶质细胞作为细胞模型，将细胞分为对照组、QC组、HIV-1 gp120组和梯度剂量QC治疗组。免疫荧光法观察应激颗粒（SGs）形成。氧化应激标志物水平和相关蛋白表达水平分别使用特异性试剂盒和Western blotting测定。HIV-1 gp120转基因小鼠（Gp120 tgm）灌胃50 mg/kg QC治疗4周。随后进行行为学评价实验，取血清进行氧化应激指标检测，取脑组织进行免疫组化染色与Western blotting分析。结果体外实验中，QC减少星形胶质细胞中的SGs（P<0.05），提高抗氧化指标水平（P<0.05），同时降低氧化物质水平（P<0.001），并且上调与SGs解聚相关的蛋白水平（P<0.05）。在体内，QC改善了Gp120 tgm的认知功能障碍表现，缓解氧化应激（P<0.05），促进SGs解聚相关蛋白表达（P<0.05）。结论 QC通过抑制氧化应激，促进SGs解聚蛋白表达和SGs解聚，减轻HIV-1 gp120诱导的星形胶质细胞毒性和改善认知功能。提示QC具备作为HIV-1相关神经认知障碍的治疗药物的潜力。

关键词: 槲皮素, 星形胶质细胞, 应激颗粒, 氧化应激, HIV-1 gp120

Abstract:

Objective To explore the effect of quercetin for mitigating HIV-1 gp120-induced astrocyte neurotoxicity and its underlying mechanism. Methods Primary rat astrocytes were isolated and treated with quercetin, HIV-1 gp120, or gradient concentrations of quercetin combined with HIV-1 gp120. The formation of stress granules (SGs) in the treated cells was observed with immunofluorescence assay, and the levels of oxidative stress markers and protein expressions were measured using specific assay kits and Western blotting. HIV-1 gp120 transgenic mice were treated with quercetin (50 mg/kg) by gavage for 4 weeks, and the changes in cognitive functions and oxidative stress levels were examined by behavioral assessments, oxidative stress index analysis in serum, and immunohistochemical and Western blotting of the brain tissue. Results In primary rat astrocytes, treatment with quercetin significantly reduced HIV-1 gp120-induced SG formation, increased the levels of antioxidant indexes, decreased the levels of oxidative substances, and up-regulated protein level associated with SG depolymerization. In the transgenic mouse models, quercetin obviously improved the cognitive function of the rats, reduced oxidative stress levels, and promoted the expression of proteins associate with SG depolymerization in the brain tissues. Conclusion Quercetin mitigates HIV-1 gp120-induced astrocyte neurotoxicity and cognitive function impairment by inhibiting oxidative stress, enhancing expressions of SG depolymerization-related proteins, and promoting SG disassembly, suggesting the value of quercetin as a potential therapeutic agent for neuroprotection in HIV-associated neurocognitive disorders.

Key words: quercetin, astrocyte, stress granules, oxidative stress, HIV-1 gp120

黄鹏伟, 陈洁, 邹金虎, 高雪锋, 曹虹. 槲皮素促进应激颗粒G3BP1解聚改善HIV-1 gp120诱导的星形胶质细胞神经毒性[J]. 南方医科大学学报, 2025, 45(2): 304-312.

Pengwei HUANG, Jie CHEN, Jinhu ZOU, Xuefeng GAO, Hong CAO. Quercetin mitigates HIV-1 gp120-induced rat astrocyte neurotoxicity via promoting G3BP1 disassembly in stress granules[J]. Journal of Southern Medical University, 2025, 45(2): 304-312.

图/表 6

图1 QC与HIV-1 gp120对星形胶质细胞活力的影响

Fig.1 Effects of concentration gradient of quercetin (QC) alone (A) and in combination with HIV-1 gp120. (B) on viability of primary rat astrocytes. ***P<0.001 vs 0 μmol/L. **P<0.01, ***P<0.001 vs HIV-1 gp120+0 μmol/L QC.

图2 QC降低星形胶质细胞中SGs阳性细胞比例

Fig.2 QC reduces the ratio of stress granule (SG)‑positive cells in HIV-1 gp120-induced primary rat astrocytes. A: Immunofluorescence staining of primary rat astrocytes. B: Ratio of SG-positive cells. C: Immunofluorescence intensity of G3BP1. D, E: Results of Western blotting for detecting G3BP1 protein expression level (fold of control). *P<0.05, **P<0.01, ***P<0.001 vs HIV-1 gp120 group.

图3 QC减轻氧化应激并促进原代星形胶质细胞SGs解聚蛋白表达

Fig.3 QC reduces oxidative stress and promotes expression of SG-depolymerizing proteins in primary rat astrocytes in vitro. A: Specific activity of SOD and CAT and contents of GSH, H2O2 and MDA in primary rat astrocytes in each group. B: ROS level and fluorescence intensity. C: Western blotting for detecting expressions levels of SG-depolymerizing proteins in primary rat astrocytes. *P<0.05, **P<0.01, ***P<0.001 vs HIV-1 gp120 group. &P<0.05, &&P<0.01.

图4 QC对Gp120 tgm脑组织氧化应激和SGs解聚蛋白表达的影响

Fig.4 Effects of QC on oxidative stress indexes and expressions of SG-depolymerizing proteins in brain tissues of Gp120 tgm mice. A: Serum levels of SOD and CAT activities and GSH, H2O2 and MDA contents in each group. B: Results of Western blotting for detecting the expression levels of SG-depolymerizing proteins in the brain tissues of the mice in each group. *P<0.05, **P<0.01, ***P<0.001 vs Gp120 tgm group. &P<0.05.

图5 QC降低Gp120 tgm脑组织SGs水平

Fig.5 QC decreases the level of SGs in the brain tissues of Gp120 tgm mice. A, B: Immunohistochemical staining for G3BP1 in the hippocampus and cortex. C, D: Results of Western blotting for detecting the expression level of G3BP1. **P<0.01, ***P<0.001 vs Gp120 tgm group.

图6 QC改善Gp120 tgm的认知功能障碍表现

Fig.6 QC improves cognitive function of Gp120 tgm mice. A-E: Results of Morris water maze test (A-C) and open field test (D-I) in each group. *P<0.05, **P<0.01, ***P<0.001 vs Gp120 tgm group.

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