Protective Effect of Tiaogeng Decoction in Improving Apoptosis of Hypothalamic Neurons Based on JNK Pathway
10.13422/j.cnki.syfjx.20192037
- VernacularTitle: 基于JNK通路探讨调更汤对GT1-7下丘脑神经元细胞凋亡的保护作用
- Author:
Sheng-nan LI
1
;
Yang LIU
1
;
Xiao-fei LIU
1
;
Chao CONG
1
;
Hui-cong LIU
1
;
Wen-jia GUI
1
;
Yue-jiao WANG
1
;
Shan XIAO
1
;
Xin SHAN
1
;
Lian-wei XU
1
Author Information
1. Longhua Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
- Publication Type:Research Article
- Keywords:
menopause syndrome;
hypothalamic neuronal apoptosis;
Tiaogeng decoction;
c-Jun N-terminal kinase pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(20):49-55
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the mechanism of Tiaogeng decoction in improving apoptosis of hypothalamic neurons through c-Jun N-terminal kinase (JNK) pathway. Method:The GT1-7 cell line of hypothalamic neuron cells treated by tributyltin chloride (TBTC) was used to induce a model of neuronal apoptosis, with 17β-estradiol as a positive drug. They were divided into control group, model group (1 mg·L-1), 17β-E2 group(100 nmol·L-1) and low, middle and high-dose Tiaogeng decoction groups (31.25,62.5,125 mg·L-1). The cell morphology was observed under a fluorescent inverted microscope, and cell counting kit-8 (CCK-8) was used to detect the cell viability. The cell nuclear morphology and the apoptosis rate of hypothalamic neurons were detected by Hoechst 33258 and Annexin V-FITC/propidium iodide(PI) double staining. Western blot was used to detect the expression levels of apoptosis signal-regulating kinase 1(ASK1), JNK, p53 and cleaved Caspase-3 of JNK pathway in GT1-7 cell. Western blot was used to analyze apoptosis-associated protein apoptosis signal-regulated kinase 1 (ASK1), JNK, p53, cleaved Caspase-3 expressions in JNK pathway and phosphorylation of JNK after intervention with JNK inhibitor SP600125, protein expression level of cleaved Caspase-3. Result:Compared with normal control group, the cell viability of the model group was significantly decreased (P<0.01), whereas the apoptotic rate was significantly increased (P<0.01). The expressions of phosphorylated ASK1(p-ASK1), p-JNK, phosphorylated p53 (p-p53) and cleaved Caspase-3 increased significantly (P<0.01). After intervention with Tiaogeng decoction, compared with model group, the activity of GT1-7 cells was significantly improved(P<0.01), while the apoptotic rate was decreased (P<0.01). The expressions of p-ASK1, p-JNK, p-p53, and cleaved Caspase-3 proteins were down-regulated. High-dose Tiaogeng decoction combined with SP600125(10 μmol·L-1)treatment significantly inhibited the expressions of p-JNK and cleaved Caspase-3 in GT1-7 cells (P<0.01). Conclusion:Tiaogeng decoction can improve the apoptosis of hypothalamic neurons through JNK pathway, and provide a theoretical basis for the treatment of menopausal syndrome and central nervous system protection.
