背景:创伤性脑损伤(TBI),特别是TBI后的神经炎症持续很长时间,并导致明显的神经退行性病变和神经精神问题。
目的:在本研究中,研究了安功牛黄(AGNH)对TBI的神经保护作用,并通过整合多个组学揭示了其机制。
方法:对TBI大鼠连续5天给予AGNH,并使用改良的神经系统严重程度评分(mNSS)评估其效果。脑水肿,H&E染色,Nissl染色和TUNEL染色。通过使用RNA测序(RNA-seq)和代谢组学分析揭示了该机制。炎症因子,通过酶联免疫吸附试验验证凋亡相关蛋白和确定的重要靶标,免疫印迹和免疫荧光染色。
结果:AGNH的管理降低了mNSS,脑水肿,大脑结构损伤,但TBI大鼠的Nissl体密度增加。此外,AGNH降低IL-1β,IL-17A,TNF-α,MMP9,MCP-1,IL-6,Bax和TUNEL染色,但Bcl2水平升高。整合转录组学分析和代谢组学分析确定了重要靶标和关键代谢途径。重要的是,AGNH处理降低TLR4、MYD88、NLRP3、BTK、IL-18和Caspase1以及甘油磷脂代谢相关蛋白AGPAT2和PLA2G2D,并降低了TBI大鼠脑内NF-κBp65的核转位。此外,AGNH增加磷脂酰胆碱(PC),磷脂酰甘油(PG),磷脂酰丝氨酸(PS),磷脂酰乙醇胺(PE),但是减少了甘油磷脂代谢代谢途径中的1-酰基-sn-甘油-3-磷酸胆碱(LysoPC)。
结论:综合来看,AGNH抑制NF-κB/NLRP3轴以抑制神经炎症,细胞凋亡和焦亡,改善了TBI后甘油磷脂代谢的代谢途径。
BACKGROUND: Traumatic brain injury (TBI), especially neuroinflammation after TBI persists for a long time and causes significant neurodegenerative pathologies and neuropsychiatric problems.
OBJECTIVE: In this study, the neuroprotective effect of AnGong NiuHuang (AGNH) on TBI was investigated and the mechanism was revealed by integrating multiple omics.
METHODS: The rats with TBI were administrated with AGNH for 5 consecutive days and the effect was evaluated by using modified neurologic severity score (mNSS), brain edema, H&E staining, Nissl staining and TUNEL staining. The mechanism was revealed by using RNA sequencing (RNA-seq) and metabolomic analysis. The inflammatory factors, apoptosis-related proteins and identified vital targets were validated by enzyme-linked immunosorbent assay, western blotting and immunofluorescence staining.
RESULTS: Administration of AGNH decreased mNSS, brain edema, brain structure damage, but increased Nissl body density in the rats with TBI. Additionally, AGNH reduced IL-1β, IL-17A, TNF-α, MMP9, MCP-1, IL-6, Bax and TUNEL staining,but elevated Bcl2 level. Integrating transcriptomic analysis and metabolomic analysis identified vital targets and critical metabolic pathways. Importantly, AGNH treatment reduced the expression of TLR4, MYD88, NLRP3, BTK, IL-18 and Caspase 1 as well as glycerophospholipid metabolism-related protein AGPAT2 and PLA2G2D, and decreased the nuclear translocation of NF-κB p65 in the brain of TBI rats. Additionally, AGNH increased phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylserine (PS), phosphatidylethanolamine (PE), but decreased 1-acyl-sn-glycero-3-phosphocholine (LysoPC) in the metabolic pathway of glycerophospholipid metabolism.
CONCLUSIONS: Taken together, AGNH inhibited NF-κB/NLRP3 axis to suppress neuroinflammation, cell apoptosis and pyroptosis, and improved metabolic pathways of glycerophospholipid metabolism after TBI.