Study on Protective Effects and Mechanism of Lespedeza cuneata Extracts on Glutamate-induced Hippocampal Cells HT 22 Injury of Mice Based on Nrf 2/HO-1 Signaling Pathway
- VernacularTitle:基于Nrf2/HO-1信号通路的夜关门提取物对谷氨酸诱导小鼠海马细胞HT22损伤的保护作用及机制研究
- Author:
Feng GUO
1
;
Shan HUANG
2
;
Bin LI
2
Author Information
1. Dept. of Pharmacy，Baicheng Medical College，Jilin Baicheng 137000，China
2. Dept. of Pharmacy，Qingdao Universit y of Science and Technology，Shandong Qingdao 266000，China
- Publication Type:Journal Article
- Keywords:
Lespedeza cuneata;
Glutamate;
HO-1;
Nrf2;
Mice;
Hippocampal cells HT 22;
Mechanism
- From:
China Pharmacy
2020;31(11):1303-1308
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE：To study the protective effects of Lespedeza cunea ta extract on glutamate-induced hippocampal cells HT22 injury of mice and its possible mechanism based on Nrf 2/HO-1 signaling pathway. METHODS ：Using glutamate （5 mmol/L） to extablish the injury model of HT 22 cells. Using water soluble vitamin E as positive control （50 µmol/L），MTT assay was used to detect the effects of 0（blank control ），25，50，100 µg/mL petroleum ether extract ，dichloromethane extract ，ethyl acetate extract of L. cuneata on the proliferation of glutamate-induced injury cellsafter pretreated for 12 h. Using water soluble vitamin E as positive control （50 µmol/L），DCFH-DA assay was used to detect the effects of 0（blank control ），25，50，100 µg/mL L. cuneata dichloromethane extract on the level of active oxygen （ROS）in glutamate-induced injury cells after pretreated with 12 h. Using HO-1 agonist CoPP as positive control ，Western blotting method was used to detect the effects of 0（blank control ），25，50，100 µg/mL L. cuneata dichloromethane extract on the protein expression of HO- 1 after treated for 24 h. Western blotting method （treated for 0.5，1，1.5 h）and immunofluorescence staining （treated for 1 h）were used to detect the effects of 100 µg/mL L. cuneata dichloromethane extract on protein expression of Nrf 2 inside and outside the nucleus. After HO-1 gene was silenced by small interfering RNA （Si RNA ）transfection technology ，the effects of 100 µg/mL L. cuneata dichloromethane extract on the survival rates of glutamate-induced injury cells and the level of ROS were detected. RESULTS ：Compared with blank control ，50， 100 µg/mL L. cuneata dichloromethane extract could significantly improve the survival rate of glutamate-induced injury cells （P＜ 0.05），while reduced the level of ROS （P＜0.05）. 25，50， 100 µg/mL L. cuneata dichloromethane extract could increase the protein expression of HO- 1 in cells（P＜0.05），while 100 com µg/mL L. cuneata dichloromethane extract could significantly decrease the protein le vel of Nrf 2 in cytoplasm and increasethat in nucleus （P＜0.05）. After HO-1 gene silencing ，the effects of L. cuneata dichloromethane extract on the proliferation promotion of glutamate-induced injury cells and the reduction of ROS level were reversed （P＜0.05）. CONCLUSIONS ：L. cuneata dichloromethane extract can protect HT 22 cells against injury induced by glutamate through activating Nrf 2 pathway，inducing HO- 1 expression.
