Molecular Mechanism of Shengxiantang in Treating Myasthenia Gravis： Based on Network Pharmacology and Molecular Docking / 中国实验方剂学杂志

ObjectiveTo predict the pharmacodynamic basis and core target of Shengxiantang in the treatment of myasthenia gravis （MG） by network pharmacology and molecular docking and to further verify the molecular mechanism through animal experiment. MethodThe active components and potential targets of Shengxiantang were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， and the disease-related targets from GeneCards and other databases. Then the common targets of the decoction and the disease were screened out， followed by the construction of protein-protein interaction （PPI） network， and Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment of the common targets based on STRING database and Cytoscape 3.8.2. Afterward， Cytoscape 3.8.2 was employed to construct the disease-active component-target network. AutoDock and PyMOL were used for molecular docking of key components and hub genes. Finally， we used the Rα97-116 peptide to induce experimental autoimmune myasthenia gravis （EAMG） in rats and then verified the core target yielded in the docking with the model rats. ResultA total of 655 disease-related targets， 118 active components of the decoction， 21 common targets of the disease and the decoction， and 3 hub genes were screened out. The common targets were mainly involved in the GO terms of regulation of active oxygen metabolism， positive regulation of protein transport， and positive regulation of protein localization， and the KEGG pathways of toll-like receptor signaling pathway， phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway， hypoxia inducible factor-1 （HIF-1） signaling pathway， and T cell receptor signaling pathway. The results of molecular docking showed that quercetin and Akt1 had the lowest and stable binding energy and interacted with each other through the amino acid residue LYS-30. Western blot demonstrated that Shengxiantang significantly inhibited the expression of p-Akt protein in the spleen of EAMG rats. ConclusionThe pharmacological mechanism of Shengxiantang in the treatment of MG may be that the main chemical components regulate the expression of the core protein Akt， and then may participate in and affect PI3K/Akt signaling pathways， laying a theoretical and experimental basis for further research.

Butyrate protects liver against ischemia reperfusion injury by inhibiting nuclear factor kappa B activation in Kupffer cells.

BACKGROUND: The inflammatory response after hepatic ischemia reperfusion (I/R) contributes to liver dysfunction and failure after transplantation. Butyrate is a four-carbon fatty acid, normally produced by bacterial fermentation of fiber in mammalian intestines, with anti-inflammatory activities. The purpose of the present study was to investigate the protective effect of butyrate preconditioning, if any, against hepatic I/R injury in rats and the underlying mechanisms involved. METHODS: Male Sprague-Dawley rats were subjected to a partial (70%) hepatic ischemia for 60 min after pretreatment with either vehicle or butyrate, followed by 3, 6, and 24 h of reperfusion. Hepatic injury was evaluated by biochemical and histopathologic examinations. Neutrophil infiltration was measured by myeloperoxidase (MPO) activity. The expression of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) was measured by enzyme-linked immunosorbent assay (Elisa) and Real-time reverse-transcriptase polymerase chain reaction (RT-PCR). The expression of nuclear factor kappa B (NF-κB) p65 was determined by immunohistochemistry and Western blot analysis. RESULTS: Butyrate treatment markedly improved hepatic function and histology, as indicated by reduced transaminase levels and ameliorated tissue pathologic changes. The expression of tumor necrosis factor-alpha, interleukin-6, and myeloperoxidase activity was attenuated by butyrate. Butyrate also reduced I/R-induced nuclear translocation of NF-κB p65 in Kupffer cells. CONCLUSION: Our results suggest that butyrate alleviates I/R-induced liver injury, possibly by suppressing inflammatory factors production and preventing NF-κB activation in Kupffer cells.

[Effect of ambroxol on lung development of rat models with nitrofen-induced congenital diaphragmatic hernia].

OBJECTIVE: To investigate the effects of ambroxol on rat models with nitrofen-induced congenital diaphragmatic hernia (CDH) and its potential mechanism. METHODS: Nine pregnant female SD rats were randomly divided into 3 groups at Day 9.5: 2 ml olive oil intragastrically in control group (2 rats) and 200 mg nitrofen in nitrofen (2 rats) and ambroxol groups (5 rats). Antenatal ambroxol was given intraperitoneally to ambroxol group at Days 18.5, 19.5 and 20.5 of gestation while control and nitrofen groups only received intraperitoneal normal saline. At Day 21.5 the fetuses were delivered by cesarean section. Incidence of hernia, lung weight/body weight (LW/BW), mean terminal branch density (MTBD), percentage of lung alveolar area (PLAA), percentage of wall thickness (MT%) and the expression of TGF-ß1 were observed. RESULTS: There were 19 CDH fetuses in nitrofen group (68.4%). The incidence of hernia in ambroxol group was 65.1% (28/43). There was no significant difference (P>0.05) between two groups. LW/BW and PLAA decreased while MTBD and MT% increased significantly in the nitrofen group versus the control group [(45±6) mg/g vs (60±7) mg/g, (50.1±4.0)% vs (58.4±3.0)%, (14.0±1.8) vs (8.5±1.1), (45±6)% vs (29±6)%, all P=0.001]. After ambroxol intervention, the ambroxol group showed a higher PLAA but a lower MTBD and MT% [(54.0±2.0)%, (12.2±2.1), (39±4)%] than those in the nitrofen group (P=0.001, 0.006, 0.002). The expression of TGF-ß1 in pulmonary tissues of the nitrofen group was significantly higher than that in the control group (13,594±3113 vs 9447±1355, P=0.001). It decreased after ambroxol intervention (10 015±818, P=0.01). CONCLUSION: Though with no effect upon the occurrence of CDH in rats, the administration of ambroxol may improve the pulmonary maturity. The down-regulated expression of TGF-ß1 and the oxidative stability are possible mechanisms.