Effects of Modified Naotai Prescription on Inflammatory Pathway of SIRT1/NF-κB in Hypoxia/Reoxygenation Injured Hippocampal Neuron

YI Yaqiao, LIU Jian, LIU Lin, CHENG Shaowu, LIAO Jun, WANG Guozuo, TAN Hu, LIU Jiyong, CHEN Junwei, WANG Wei, GUO Yanxing, GE Jinwen

YI Yaqiao1, LIU Jian2, LIU Lin2, CHENG Shaowu1, LIAO Jun3, WANG Guozuo1, TAN Hu1,

LIU Jiyong1, CHEN Junwei1, WANG Wei1, GUO Yanxing4, GE Jinwen3

1. Hunan University of Chinese Medicine, Changsha 410208, China;

2. First Hospital of Hunan University of Chinese Medicine, Changsha 410007, China;

3. Integrative Medicine Basic Key Disciplines of Hunan Province, Hunan University of Chinese Medicine, Changsha

410208, China; 4. Luoyang Bonesetting Hospital of Henan Province, Luoyang 471002, China

Abstract: Objective To observe the effects of modified Naotai Prescription on inflammatory pathway of

SIRT1/NF-κB in hypoxia/reoxygenation injured hippocampal neurons; To explore the mechanism of action. Methods Hippocampal neurons from SD rats were primitively isolated, cultured and identified. Hypoxia/ reoxygenation (H/R) cell model was established by hypoxia 24 h and reoxygenation 2 h intervention. The cultured cells were randomly divided into normal group, blank serum group, model group, positive medicine serum group and

modified Naotai Prescription group. Normal group and model group were given same amount culture medium, while other groups were given relevant amount of 10% medicated serum or blank serum to continue for reoxygenation and incubate for 2 h. Hippocampal neuronal cell viability was detected by MTT assay. The expressions of SIRT1, IκBα and NF-κB in hippocampal neurons were detected by HCA. Results Compared with the normal group, hippocampal neuronal cell viability significantly decreased (P<0.01), the neural network and dendrites and dendritic spines were broken or reduced, the cells were obviously damaged, and the expressions of SIRT1 and IκBα protein in neurons significantly decreased (P<0.05), and the expression of NF-κB protein significantly increased (P<0.05) in the model group. Compared with the model group, hippocampal neuronal cell viability significantly increased (P<0.05), the neural network and dendritic spine were significantly improved, and the expressions of SIRT1 and IκBα protein in the neurons increased (P<0.05), and the expression of NF-κB protein was down-regulated (P<0.05) in positive medicine serum group and modified Naotai Prescription group. Conclusion Modified Naotai Prescription has a significant regulatory effect on the inflammatory-related signaling pathway SIRT1/NF-κB in hippocampal neurons after hypoxia/reoxygenation injury, which may be an important mechanism for protecting hypoxic/reoxygenated hippocampal neurons and then anti-vascular dementia.

Keywords: modified Naotai Prescription; hippocampal neurons; hypoxia/reoxygenation; SIRT1; IκBα; NF-κB; vascular dementia; rats