Correlation of Autophagy with Tumor Immune Escape and Intervention with Chinese Medicine： A Review / 中国实验方剂学杂志

Autophagy and tumor immune escape are important biological mechanisms in the process of tumor cell proliferation and metastasis， involving multiple signaling pathways. The interaction of autophagy and tumor immune escape seriously affects the treatment and prognosis of tumor diseases. However， the correlation between autophagy and tumor immune escape is still not fully elucidated. Recent studies have shown that autophagy can affect the activity of immune cells by regulating the presentation of antigens in tumor cells， the release of cytokines， and the degradation of immune checkpoint proteins， thereby positively or negatively regulating tumor cell immune escape. The activation of autophagy in tumor cells can inhibit the activation of the innate immune sensing pathway of stimulator of interferon genes （STING）-type Ⅰ interferon （IFN-Ⅰ） to inhibit its immunogenicity and cytotoxic T lymphocytes （CTLs）， which promotes tumor immune escape. While autophagy suppression can reduce the infiltration of M2 macrophages， promote the binding of natural killer group 2， member D （NKG2D） to its ligand， and inhibit the recognition of immune checkpoint proteins， thereby exerting an immune-killing effect and inhibiting tumor immune escape. Traditional Chinese medicine （TCM） has unique advantages in anti-tumor research， especially in the unilateral regulation of autophagy or improvement of tumor immunity， but the research based on the regulation of autophagy and tumor immunity by TCM is insufficient. A few studies have shown that Chinese medicine monomers and compounds can exert an anti-tumor effect by regulating cell autophagy and interfering with tumor immune escape， but there is still a lack of systematic elaboration. The present study reviewed correlation between autophagy and tumor immune escape and regulation of autophagy by Chinese medicine to interfere with tumor immune escape to provide new ideas for research on mechanism of TCM against tumor diseases and development of innovative TCM drugs against tumors.

Molecular Mechanism of Qinghuangsan Against Acute Myeloid Leukemia Based on Network Pharmacology and In Vitro Experiments / 中国实验方剂学杂志

ObjectiveTo predict the therapeutic target genes and related signaling pathways of Qinghuangsan （QHP） in the treatment of acute myeloid leukemia （AML） by network pharmacology，molecular docking，and further clarify its mechanisms through in vitro cell experiment. MethodThe active components and targets of QHP were retrieved from traditional Chinese medicine systems pharmacology database and analysis platform （TCMSP），traditional Chinese medicine integrated database （TCMID），TargetNet and SwissTargetPrediction databases，and AML-related target genes were obtained by GeneCards and online mendelian inheritance in man （OMIM） databases. After screening the common targets of QHP and AML，the protein-protein interaction （PPI） network of the common targets was constructed with STRING，followed by gene ontology （GO） term and Kyoto encyclopedia of genes and genomes （KEGG） pathway enrichment analysis based on RStudio software and clusterProfiler，Bioconductor packages. At the same time，Cytoscape software is used to construct the network of "disease-component-target" and "compound-target-pathway". Select the active ingredients of QHP for molecular docking with the top 8 targets in the "compound-target-pathway" network. In vitro cell experiment and Western blot were used to further verify the anti-AML effect of QHP. ResultThe prediction results show that there are 11 main active components of QHP，and 22 common targets of QHP and AML are collected. KEGG pathway analysis results show that phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） and mitogen-activated protein kinase （MAPK） signaling pathways may play a key role in the treatment of AML disease by QHP. "Compound-target-pathway" network analysis showed that the top 8 targets include Akt1，phosphatidylinositol-4，5-bisphosphate 3-kinase catalytic subunit alpha （PIK3CA），mitogen-activated protein kinase kinase 1 （MAP2K1），TP53，serine/threonine kinase （RAF1），B cell lymphoma（Bcl）-2，cysteine aspartic acid specific protease（Caspase）-9 and JUN. Molecular docking results showed that 3-indolyl-β-D-glucopyranoside was optimally docked with MAP2K1，isovitexin docked with PIK3CA，and indirubin docked with Bcl-2. Cell experiments show that 3-indolyl-β-D-glucopyranoside，isovitexin and indirubin can effectively inhibit the proliferation of AML cells，regulate the MAPK/PI3K signaling pathway，and inhibit the expression of Bcl-2 protein. ConclusionQHP can treat AML through "multi-component，multi-target，multi-pathway" synergistic treatment，and its mechanism of pharmacology may be related to the regulation of MAPK signaling pathway and PI3K/Akt signaling pathway.

Effects of total C-21 steroidal glycosides from Baishouwu on proliferation invasion and apoptosis of human hepatic cancer HepG2 cells / 中国中药杂志

This study aimed to investigate the inhibitory effect of total C-21 steroidal glycoside (TCSG) from Baishouwu on the proliferation, invasion and apoptosis of human hepatoma HepG2 cells in vitro and the relevant molecular mechanism. The experiment was divded into control group, TCSG groups (25, 60, 150 mg·L⁻¹) and positive control cisplatin group (1.33 mg·L⁻¹). Human hepatocyte L-02 cells and hepatoma HepG2 cells were treated with different concentrations of TCSG. Then, the inhibitory effect of TCSG on the proliferation of HepG2 cells was detected by CCK-8 method. Cell cycle, cell apoptosis and mitochondrial membrane potential were detected by flow cytometry. The apoptotic morphology was observed by Hoechst 33258 staining. Cell migration and invasion abilities were analyzed by Transwell chamber model. The protein expressions of Bcl-2, Bax, caspase 3, cleaved caspase 3 and Cyt C (cytosolchondrial) were detected by Western blot. Compared with the control group, the proliferation of HepG2 cells was significantly inhibited after treatment with different concentrations of TCSG for 48 h in a dose-dependent manner(<0.01), but no obvious effect was observed on the proliferation of L-02 cells. After treatment with TCSG for 48 h, apoptotic morphology such as nuclear shrinkage, fragmentation and semilunar or circular was observed; migration and invasion abilities of cells were significantly decreased, cell cycle was blocked in the G₀/G₁ phase(<0.01), mitochondrial membrane potential was remarkably decreased(<0.01), and so did the ratio of apoptosis(<0.01).Western blot results showed that the protein expressions of Bax, caspase 3, cleaved caspase 3, and Cyt C were significantly up-regulated(<0.05, <0.01), while the Bcl-2 protein was significantly down-regulated(<0.05, <0.01). Furthermore, the ratio of Bax/Bcl-2 was increased (<0.01). The results suggested that TCSG could inhibit the proliferation and invasion of HepG2 cells, and induce the apoptosis of HepG2 cells. The potential mechanism may be related to the blocking of cell cycle and the regulation of the expressions of apoptosis-related proteins by activating mitochondrial pathway.

Research progress on the correlation between TLR4-MyD88-NF-κB signalling pathways and the hepatic inflammation-fibrosis-cancer axis / 国际药学研究杂志

As a critical pathway in the reaction of hepatic disease, Toll-like receptor 4(TLR4)-myeloid differentiation factor 88(MyD88)-nuclear transcription factor(NF-κB)signalling pathway, which widely exists in various tissues and cells, is one of the most important signalling pathways that mediate the expression of inflammatory factor between the intracellular and intercellular. Study found that TLR4-MyD88-NF-κB signalling pathway plays an important role in the regulation of liver immune abnormalities, inflammatory response triggered by the liver damage, activation of hepatic stellate cells and liver fibrosis deterioration and the development of cancer of the liver. The signalling pathway may be an important regulatory point in the dynamic changes of the”hepatic inflammationfibrosis-cancer axis”(IFC axis)and liver cancer metastasis. In this paper, we review the recent advances in the pathological mechanism of TLR4-MyD88-NF-κB signalling pathway participating in IFC axis disease, so as to provide reference for related research in the future.