Neuroprotective Effect of Modified Yuejuwan and Ganmai Dazao Tang on Glutamate-induced HT22 Cell Injury Model
10.13422/j.cnki.syfjx.20190836
- VernacularTitle: 越鞠丸合甘麦大枣汤加减对谷氨酸诱导的HT22细胞损伤模型的神经保护作用
- Author:
Ke-min GE
1
;
Wei WANG
1
;
Wen-da XUE
1
;
Gang CHEN
1
;
Fu-shun WANG
1
Author Information
1. School of Medicine and Life Sciences, School of Psychology, School of Basic Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China
- Publication Type:Research Article
- Keywords:
Yuejuwan;
Ganmai Dazao Tang;
hippocampal neuron cell line (HT22);
glutamate;
neuroprotection
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(12):22-27
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the neuroprotective effect of modified Yuejuwan and Ganmai Dazao Tang (YJGZ) on glutamate-induced cell injury of mouse hippocampal neuron cell line (HT22). Method: The HT22 cell was cultured aseptically, and the cell injury model was established with high concentration of glutamate acid, YJGZ aqueous extract and drug-containing serum were prepared. Then experiment was divided into normal group, model group, YJGZ drug-containing serum groups (1%, 5%, 10%), YJGZ aqueous extract group (166 mg·L-1), and nimodipine group (100 μmol·L-1). Methye thiazolye telrazlium (MTT) was used to detect the cell survival rate of each group, the level of lactate dehydrogenase (LDH) release and intracellular reactive oxygen species (ROS) were detected by enzyme-linked immunosorbent assay (ELISA). Western blot was used to detect the protein expression of hippocampal N-methyl-D-aspartate receptor subunit 2B protein (NR2B), cyclic adenosine response element binding protein (CREB), phosphorylated cyclic adenosine response element binding protein (p-CREB), extracellular regulated protein kinase (ERK),and phosphorylated extracellular regulated protein kinase (p-ERK). Result: As compared with normal group, the cell survival rate was significantly decreased in model group (P<0.01), LDH release rate was significantly increased (P<0.05), intracellular reactive oxygen species (ROS) level was increased (P<0.05), expression of NR2B protein was increased (P<0.05), and the protein expression levels of CREB, p-CREB, ERK and p-ERK were decreased (P<0.05). As compared with model group, YJGZ aqueous extract group and nimodipine group can significantly improve the survival rate of HT22 cells under glutamate model conditions (P<0.01), reduce cell damage, reduce LDH release rate and intracellular ROS levels (P<0.05,P<0.01), decrease the expression level of NR2B protein (P<0.05), and increase the protein expression levels of CREB, p-CREB, ERK and p-ERK (P<0.05). However, the survival rate of HT22 cells was not increased in the YJGZ-containing serum group as compared with model group. Conclusion: YJGZ aqueous extract has a significant protective effect on glutamate-induced HT22 cell injury.
