Protective effect of ginsenoside Rh2 against Toxoplasma gondii infection-induced neuronal injury through binding TgCDPK1 and NLRP3 to inhibit microglial NLRP3 inflammasome signaling pathway,International Immunopharmacology

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Protective effect of ginsenoside Rh2 against Toxoplasma gondii infection-induced neuronal injury through binding TgCDPK1 and NLRP3 to inhibit microglial NLRP3 inflammasome signaling pathway
International Immunopharmacology ( IF 4.8 ) Pub Date : 2022-09-05 , DOI: 10.1016/j.intimp.2022.109176
Guang-Nan Jin ₁ , Jing-Mei Lu ₁ , Hui-Wen Lan ₁ , Yu-Nan Lu ₁ , Xin-Yu Shen ₁ , Xiang Xu ₁ , Lian-Xun Piao ₁

Affiliation

Background

Toxoplasma gondii (T. gondii) is a neurotropic obligate intracellular parasite that can activate microglial and promote neuronal apoptosis, leading to central nervous system diseases. The NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome signaling complex plays a key role in inducing neuroinflammation. Our previous studies have found that ginsenoside Rh2 (GRh2) inhibits T. gondii infection-induced microglial activation and neuroinflammation by downregulating the Toll-like receptor 4/nuclear factor-kappa B signaling pathway. However, whether GRh2 reduces T. gondii infection-induced neuronal injury through actions on microglial NLRP3 inflammasome signaling has not yet been clarified.

Methods

In this study, we employed T. gondii RH strain to establish in vitro and in vivo infection models in BV2 microglia cell line and BALB/c mice. Molecular docking, localized surface plasmon resonance assay, quantitative competitive-PCR, ELISA, western blotting, flow cytometric analysis, and immunofluorescence were performed.

Results

Our results showed that GRh2 alleviated neuropathological damage and neuronal apoptosis in cortical tissue of T. gondii-infected mice. GRh2 and CY-09 (an inhibitor of NLRP3) exhibited potent anti-T. gondii effects through binding T. gondii calcium-dependent protein kinase 1 (TgCDPK1). GRh2 decreased Iba-1 (a specific microglial marker) and NLRP3 inflammasome signaling pathway-related protein expression by binding NLRP3. Co-culture of microglia/primary cortical neurons revealed that T. gondii-induced microglial activation caused neuronal apoptosis, but GRh2 reduced this effect, consistent with the effects of CY-09.

Conclusion

Taken together, our results show that GRh2 has a protective effect against T. gondii infection-induced neuronal injury by binding TgCDPK1 and NLRP3 to inhibit NLRP3 inflammasome signaling pathway in microglia.

中文翻译：

人参皂甙Rh2通过结合TgCDPK1和NLRP3抑制小胶质细胞NLRP3炎性体信号通路对弓形虫感染所致神经元损伤的保护作用

背景

刚地弓形虫（T. gondii）是一种嗜神经专性细胞内寄生虫，可激活小胶质细胞，促进神经元凋亡，导致中枢神经系统疾病。NOD 样受体家族含有 pyrin 结构域的 3 (NLRP3) 炎性体信号复合物在诱导神经炎症中起关键作用。我们之前的研究发现，人参皂甙Rh2 (GRh2)通过下调Toll 样受体4/核因子-κB 信号通路来抑制弓形虫感染诱导的小胶质细胞活化和神经炎症。然而，GRh2 是否通过对小胶质细胞 NLRP3 炎性体信号传导的作用来减少弓形虫感染诱导的神经元损伤尚未阐明。

方法

在本研究中，我们采用弓形虫RH 菌株在 BV2 小胶质细胞系和 BALB/c 小鼠中建立体外和体内感染模型。进行了分子对接、局部表面等离子共振测定、定量竞争性PCR、ELISA、蛋白质印迹、流式细胞术分析和免疫荧光。

结果

我们的研究结果表明，GRh2 减轻了弓形虫感染小鼠皮质组织中的神经病理学损伤和神经元凋亡。GRh2 和 CY-09（NLRP3 的抑制剂）通过结合刚地弓形虫钙依赖性蛋白激酶 1 (TgCDPK1 ) 表现出有效的抗弓形虫效果。GRh2 通过结合 NLRP3 降低了 Iba-1（一种特定的小胶质细胞标志物）和 NLRP3 炎症小体信号通路相关蛋白的表达。小胶质细胞/初级皮层神经元的共培养表明，弓形虫诱导的小胶质细胞激活导致神经元凋亡，但 GRh2 降低了这种效应，与 CY-09 的作用一致。