Shentu Yangping, Chen Mengfan, Wang Hui, Du Xiaotong, Zhang Wenjing, Xie Guizhen, Zhou Shaoyan, Ding Lu, Zhu Yun, Zhu Min, Zhang Nan, Du Congkuo, Ma Jianshe, Chen Ran, Yang Jinge, Fan Xiaofang, Gong Yongsheng, Zhang Hongyu, Fan Junming
Department of Pathology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China.
Exp Neurol. 2024 Jan;371:114586. doi: 10.1016/j.expneurol.2023.114586. Epub 2023 Oct 26.
Hydrogen sulfide (HS), an endogenous gasotransmitter, exhibits the anxiolytic roles through its anti-inflammatory effects, although its underlying mechanisms remain largely elusive. Emerging evidence has documented that cell cycle checkpoint kinase 1 (Chk1)-regulated DNA damage plays an important role in the neurodegenerative diseases; however, there are few relevant reports on the research of Chk1 in neuropsychiatric diseases. Here, we aimed to investigate the regulatory role of HS on Chk1 in lipopolysaccharide (LPS)-induced anxiety-like behavior focusing on inflammasome activation in the hippocampus. Cystathionine γ-lyase (CSE, a HS-producing enzyme) knockout (CSE) mice displayed anxiety-like behavior and activation of inflammasome-mediated inflammatory responses, manifesting by the increase levels of interleukin-1β (IL-1β), IL-6, and ionized calcium-binding adaptor molecule-1 (Iba-1, microglia marker) expression in the hippocampus. Importantly, expression of p-Chk1 and γ-H2AX (DNA damage marker) levels were also increased in the hippocampus of CSE mice. LPS treatment decreased the expression of CSE and CBS while increased p-Chk1 and γ-H2AX levels and inflammasome-activated neuroinflammation in the hippocampus of mice. Moreover, p-Chk1 and γ-H2AX protein levels and cellular immunoactivity were significantly increased while CSE and CBS were markedly decreased in cultured BV2 cells followed by LPS treatment. Treatment of mice with GYY4137, a donor of HS, inhibited LPS-induced increased in p-Chk1 and γ-H2AX levels, mitigated inflammasome activation and inflammatory responses as well as amelioration of anxiety-like behavior. Notably, SB-218078, a selective Chk1 inhibitor treatment attenuated the effect of LPS on inflammasome activation and inflammatory responses and the induction of anxiety-like behavior. Finally, STAT3 knockdown with AAV-STAT3 shRNA alleviated LPS-induced anxiety-like behavior and inhibited inflammasome activation in the hippocampus, and blockade of NLRP3 with MCC950 attenuated neuroinflammation induction and ameliorated LPS-induced anxiety-like behavior. Overall, this study indicates that downregulation of Chk1 activity by HS activation may be considered as a valid strategy for preventing the progression of LPS-induced anxiety-like behavior.
硫化氢（HS）作为一种内源性气体递质，通过其抗炎作用发挥抗焦虑作用，但其潜在机制仍不清楚。新出现的证据表明，细胞周期检查点激酶1（Chk1）调节的DNA损伤在神经退行性疾病中起重要作用；然而，关于Chk1在神经精神疾病方面的研究报道较少。在此，我们旨在研究HS对Chk1在脂多糖（LPS）诱导的焦虑样行为中的调节作用，重点关注海马体中的炎性小体激活。胱硫醚γ-裂解酶（CSE，一种产生HS的酶）基因敲除（CSE）小鼠表现出焦虑样行为以及炎性小体介导的炎症反应激活，表现为海马体中白细胞介素-1β（IL-1β）、IL-6水平升高以及离子钙结合衔接分子-1（Iba-1，小胶质细胞标志物）表达增加。重要的是，CSE小鼠海马体中p-Chk1和γ-H2AX（DNA损伤标志物）水平的表达也增加。LPS处理降低了CSE和CBS的表达，同时增加了小鼠海马体中p-Chk1和γ-H2AX水平以及炎性小体激活的神经炎症。此外，LPS处理后的培养BV2细胞中，p-Chk1和γ-H2AX蛋白水平以及细胞免疫活性显著增加，而CSE和CBS则明显降低。用HS供体GYY4137处理小鼠，可抑制LPS诱导的p-Chk1和γ-H2AX水平升高，减轻炎性小体激活和炎症反应以及改善焦虑样行为。值得注意的是，选择性Chk1抑制剂SB-218078处理减弱了LPS对炎性小体激活和炎症反应的影响以及焦虑样行为的诱导。最后，用腺相关病毒-STAT3短发夹RNA敲低STAT3可减轻LPS诱导的焦虑样行为，并抑制海马体中的炎性小体激活，用MCC950阻断NLRP3可减轻神经炎症诱导并改善LPS诱导的焦虑样行为。总体而言，本研究表明，通过激活HS下调Chk1活性可能是预防LPS诱导的焦虑样行为进展的有效策略。
