Qingxin Kaiqiao Fang Inhibits Aβ 25-35-Induced Apoptosis in Primary Cultured Rat Hippocampal Neuronal Cells via the p38 MAPK Pathway: An Experimental Validation and Network Pharmacology Study.

Tian-Qi Wang,Yan Shen,Jian-Wei Lin,Shi-Yu Gao,Hai-Yan Hu,Xiao-Xiao Lai,Lu-Ting Xu

doi:10.1155/2020/9058135

Abstract

Qingxin kaiqiao fang (QKF), a traditional Chinese medicine compound, has been applied to treat Alzheimer's disease (AD) for many years and has exhibited remarkable effects. However, the underlying mechanism is still not explicit. The current study aims to investigate whether QKF exerts an antiapoptotic role through the p38 MAPK pathway in the course of AD. Network pharmacology analysis was applied to study the effective components, possible therapeutic targets, and AD-related pathway of QKF. Further, the AD cell model was established using amyloid-beta (Aβ)25-35 peptide and primary hippocampal neuronal cells extracted from newborn Sprague-Dawley rats. Microtubule-associated protein-2 (MAP-2) imaging was used to detect the morphology of hippocampal neurons. Western blot (WB) analysis was applied to detect the protein expression levels of p38 MAPK, p-p38 MAPK, Bcl-2, Bax, caspase-3, and cleaved caspase-3. Cell viability and apoptosis were determined using cell counting kit-8 (CCK-8) and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assays, respectively. SB203580 and U46619 were used to detect changes in cell morphology, cell viability, and apoptosis upon inhibiting or activating p38 MAPK. Our present work showed that QKF protects hippocampal neuronal morphology, enhances cell viability, and reduces the number of TUNEL-positive cells. In addition, our results showed that QKF increased the expression levels of antiapoptotic proteins and decreased the expression of proapoptotic proteins. QKF at 25 mg·mL−1 best inhibited neuronal apoptosis among the three doses of QKF by suppressing p38 MAPK activity. Collectively, QKF plays an antiapoptotic role via the p38 MAPK pathway.

Highlights

As a serious neurodegenerative disease in aged individuals, Alzheimer’s disease (AD) is characterized by the formation of intracellular neurofibrillary tangles, extracellular deposits of senile plaques (SP), and the loss of neurons [1]
It is generally accepted that the apoptosis-induced death of a large number of neurons is a common feature in the brains of patients with AD [2]. is programmed cell death can be induced by multiple factors, one of which is the abnormal aggregation and deposition of amyloid-beta (Aβ) fragments, the main component of SP
Qingxin kaiqiao fang (QKF), which is based on Fumanjian, a notable traditional Chinese medicine compound from Jingyue Quanshu that was first described by Zhang Jingyue during the Ming Dynasty, is made mainly from Radix Rehmanniae (Sheng Di Huang), Radix Paeoniae Alba (Bai Shao), Radix Ophiopogonis (Mai dong), Cortex Moutan Radicis (Mu dan pi), Poria cocos (Fu Ling), Herba Dendrobii Rhizoma (Shi Hu), Rhizoma Acori Tatarinowii (Shi Chang Pu), Rhizoma Anemarrhenae (Zhi mu), Sophorae Flavescentis (Ku Shen), and Pericarpium Citri Reticulatae (Chenpi)

Summary

Introduction

As a serious neurodegenerative disease in aged individuals, Alzheimer’s disease (AD) is characterized by the formation of intracellular neurofibrillary tangles, extracellular deposits of senile plaques (SP), and the loss of neurons [1]. It is generally accepted that the apoptosis-induced death of a large number of neurons is a common feature in the brains of patients with AD [2]. Is programmed cell death can be induced by multiple factors, one of which is the abnormal aggregation and deposition of amyloid-beta (Aβ) fragments, the main component of SP. Evidence-Based Complementary and Alternative Medicine hippocampal neuronal damage induced by Aβ can be mediated by the regulation of p38 mitogen-activated protein kinase (p38 MAPK) [6, 7]. P38 MAPK activation leads to programmed neuronal cell death primarily through alterations in the expression of proteins involved in apoptosis, including caspase-3, the antiapoptotic protein regulator Bcl, and the proapoptotic protein regulator Bax [10]. A network pharmacology analysis, which integrates high-throughput data integration, database retrieval, data mining, target prediction, laboratory simulations, and other research methods [13], was applied in this study to reveal the complex mechanism of QKF in the treatment of AD

Methods

Results

Conclusion