Protective Effect of Gandou Decoction on Cer Signaling Pathway in HT-22 Induced by High Copper
10.13422/j.cnki.syfjx.20190701
- VernacularTitle: 基于Cer通路探讨肝豆汤对Wilson病模型高铜诱导海马神经元细胞损伤的保护作用
- Author:
Chen-chen XU
1
;
Jian-jian DONG
1
;
Xun WANG
2
;
Yong-zhu HAN
1
;
Nan CHENG
1
Author Information
1. Hospital Affiliated to Neurological Institute, Anhui University of Chinese Medicine, Hefei 230061, China
2. The Second Affiliated Hospital of Anhui Medical University, Hefei 230601, China
- Publication Type:Research Article
- Keywords:
Wilson's disease;
Gandou decoction;
Cer signaling pathway;
HT-22 cell
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(7):56-60
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To detect ceramide(Cer) signaling pathway-related proteins expression levels in HT-22 with Gandou decoction (GDD), in order to explore its molecular targets and mechanism in regulating Cer signaling pathway. Method: The experiment was divided into normal group (normal HT-22 cultured by 10%blank rabbit serum), model group (HT-22 cells incubated with CuSO4), and GDD group (HT-22 cells incubated with CuSO4, continuously cultured by rabbit serum containing10%, 15%, 20%GDD). HT-22 cells were incubated with different concentrations of CuSO4.The cell growth and proliferation were assessed using methyl thiazolyl tetrazolium(MTT) method; flow cytometry was used to analyze the expression of reactive oxygen species (ROS); Western blot was used to detect relevant protein expression of Cer signaling pathway. Result: The results of MTT showed that CuSO4 inhibited the growth and proliferation of HT-22 cells in a time and concentration-dependent manner; flow cytometry results showed that the model group increased the release of ROS compared with the normal group (P<0.01); compared with the model group, GDD could significantly reduce the release of ROS (P<0.01). Western blot indicated that GDD concentration-dependently decreased the expressions of acid sphingomyelinase(ASM), Cer, p38 mitogen-activated protein kinase(p38 MAPK),cytochrome C(Cyt C), cysteinyl aspartate specific proteinase(Caspase) -9, Caspase-3 in HT-22 compared with the model group (P<0.05,P<0.01). Conclusion: High copper can induce oxidative stress and deactivate Cer signaling pathway, which led to hippocampal neuron injury. These findings suggest that GDD reduces neurotoxicity induced by copper overload by increasing the copper excretion that inhibits the expressions of ASM, Cer, p38 MAPK, Cyt C, Caspase-9, Caspase-3.GDD reduces neurotoxicity induced by copper overload by decreasing copper levels in brain and then regulating Cer signaling pathway.
