Ma Chongyang, Zhao Mengpei, Du Yuqiong, Jin Shuang, Wu Xiaoyi, Zou Haiyan, Zhang Qiuyun, Gao Lianyin
School of Traditional Chinese Medicine, Capital Medical University, Beijing 10069, China.
Kaifeng Second Hospital of Traditional Chinese Medicine, Kaifeng 475000, China.
Evid Based Complement Alternat Med. 2021 Mar 3;2021:6661667. doi: 10.1155/2021/6661667. eCollection 2021.
Qingdu Decoction (QDD) is a traditional Chinese medicine formula for treating chronic liver injury (CLI). . A network pharmacology combining experimental validation was used to investigate potential mechanisms of QDD against CLI. We firstly screened the bioactive compounds with pharmacology analysis platform of the Chinese medicine system (TCMSP) and gathered the targets of QDD and CLI. Then, we constructed a compound-target network and a protein-protein interaction (PPI) network and enriched core targets in Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathways. At last, we used a CLI rat model to confirm the effect and mechanism of QDD against CLI. Enzyme-linked immunosorbent assay (ELISA), western blot (WB), and real-time quantitative polymerase chain reaction (RT-qPCR) were used.
48 bioactive compounds of QDD passed the virtual screening criteria, and 53 overlapping targets were identified as core targets of QDD against CLI. A compound-CLI related target network containing 94 nodes and 263 edges was constructed. KEGG enrichment of core targets contained some pathways related to CLI, such as hepatitis B, tumor necrosis factor (TNF) signaling pathway, apoptosis, hepatitis C, interleukin-17 (IL-17) signaling pathway, and hypoxia-inducible factor (HIF)-1 signaling pathway. Three PPI clusters were identified and enriched in hepatitis B and tumor necrosis factor (TNF) signaling pathway, apoptosis and hepatitis B pathway, and peroxisome pathway, respectively. Animal experiment indicated that QDD decreased serum concentrations of alanine aminotransferase (ALT), aspartate aminotransferase (AST), endotoxin (ET), and IL-17 and increased prothrombin time activity (PTA) level. WB and RT-qPCR analyses indicated that, compared with the model group, the expression of cysteinyl aspartate specific proteinase-9 (caspase-9) protein, caspase-3 protein, B-cell lymphoma-2 associated protein (Bax) mRNA, and cytochrome c (Cyt c) mRNA was inhibited and the expression of B-cell lymphoma-2 (Bcl-2) mRNA was enhanced in the QDD group.
QDD has protective effect against CLI, which may be related to the regulation of hepatocyte apoptosis. This study provides novel insights into exploring potential biological basis and mechanisms of clinically effective formula systematically.
清毒汤（QDD）是一种用于治疗慢性肝损伤（CLI）的中药方剂。采用网络药理学结合实验验证的方法来研究QDD治疗CLI的潜在机制。我们首先利用中药系统药理学分析平台（TCMSP）筛选生物活性化合物，收集QDD和CLI的靶点。然后，构建化合物-靶点网络和蛋白质-蛋白质相互作用（PPI）网络，并在京都基因与基因组百科全书（KEGG）信号通路中富集核心靶点。最后，我们使用CLI大鼠模型来证实QDD治疗CLI的效果和机制。采用酶联免疫吸附测定（ELISA）、蛋白质印迹法（WB）和实时定量聚合酶链反应（RT-qPCR）。
QDD的48种生物活性化合物通过虚拟筛选标准，53个重叠靶点被确定为QDD治疗CLI的核心靶点。构建了一个包含94个节点和263条边的化合物-CLI相关靶点网络。核心靶点的KEGG富集包含一些与CLI相关的通路，如乙型肝炎、肿瘤坏死因子（TNF）信号通路、凋亡、丙型肝炎、白细胞介素-17（IL-17）信号通路和缺氧诱导因子（HIF）-1信号通路。确定了三个PPI簇，分别在乙型肝炎和肿瘤坏死因子（TNF）信号通路、凋亡和乙型肝炎通路以及过氧化物酶体通路中富集。动物实验表明，QDD降低了血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、内毒素（ET）和IL-17的浓度，并提高了凝血酶原时间活性（PTA）水平。WB和RT-qPCR分析表明，与模型组相比，QDD组半胱氨酸天冬氨酸特异性蛋白酶-9（caspase-9）蛋白、caspase-3蛋白、B细胞淋巴瘤-2相关蛋白（Bax）mRNA和细胞色素c（Cyt c）mRNA的表达受到抑制，而B细胞淋巴瘤-2（Bcl-2）mRNA的表达增强。
QDD对CLI具有保护作用，这可能与调节肝细胞凋亡有关。本研究为系统探索临床有效方剂的潜在生物学基础和机制提供了新的见解。
