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Protective Effects of Jin Bai Mei Yan Prescription on Oxidative Damage and Photoaging Induced by Ultraviolet B in HaCaT Cells
NORIKO Minamisawa, MIAO Ming-San, KANG Le, LIU Hui-Juan, CUI Lin-Lin, HIROKI Kanemisu, TERUMI Naito, HIROAKI Shinotsuka
2020, 3(2): 57-66. doi: 10.1016/j.dcmed.2020.06.001
[Abstract](96) [FullText HTML] (44) [PDF 949KB](2)
ObjectiveUltraviolet B (UVB) mainly acts on the skin epidermis, causing oxidative damage and apoptosis of keratinocytes. Jin Bai Mei Yan Prescription (JBMYP) comprises a variety of antioxidant traditional Chinese medicines (TCM). In this study, we aimed to evaluate the effects of JBMYP on the oxidative damage induced by UVB in human immortalized epidermal keratinocytes (HaCaT) cells.MethodsHaCaT cells were divided into six groups: control group, model (UVB) group, positive (UVB + vitamin E) group, UVB + JBMYP low dose group (160 μg/mL), UVB + JBMYP mode-rate dose group (800 μg/mL), and UVB + JBMYP high dose group (1 600 μg/mL). HaCaT cells were irradiated with UVB and treated with JBMYP for 24 h. Methyl thiazolyl tetrazolium (MTT) assay and real-time unlabeled cell function analyzer were used to assess the cell survival and proliferation rates, respectively. At the same time, the levels of intracellular reactive oxygen species (ROS), lipid peroxide malondialdehyde (MDA), glutathione (GSH), and hydroxyproline (HYP), as well as the activities of antioxidant enzyme superoxide dismutase (SOD) and catalase (CAT) were evaluated using enzyme linked immunosorbent assay (ELISA).ResultsCompared with the model group, the survival rate of HaCaT cells in each dosage group of JBMYP was significantly improved (P < 0.05). Further, JBMYP could promote the proliferation of HaCaT cells, leading to a reduction in the contents of MDA and ROS, and increase in the contents of SOD, CAT, GSH and HYP in HaCaT cells.ConclusionsJBMYP has enhanced protective effect on oxidative damage induced by UVB in HaCaT cells.
STAT3 Inhibition by Centipede Scolopendra Extract in Liver Cancer HepG2 Cells and Orthotopic Mouse Models of Hepatocellular Carcinoma
TENG Yong-Jie, LIU Zhuo, LIAO Liu, CHEN Yuan, HUANG Xiao-Di, TIAN Xue-Fei
2020, 3(2): 67-79. doi: 10.1016/j.dcmed.2020.06.002
[Abstract](104) [FullText HTML] (48) [PDF 1037KB](0)
ObjectiveTo observe the effects of Centipede Scolopendra extraction (CSE) on human liver cancer HepG2 cells and the nude mouse tumor model of liver orthotopic transplantation, and to explore the anti-liver cancer mechanism of the extract.MethodsHepG2 cells were respectively treated with CSE250 (250 μg/mL), CSE500 (500 μg/mL) and 5-FU, and control group was established. An enzymatic hydrolysis and acetone precipitation method was used to separate and purify CSE, which was then used to treat HepG2 cells. The CCK8 assay was used to detect the inhibition of cell proliferation and the half maximal inhibitory concentration (IC50) was calculated. Flow cytometry was used to analyze the cell cycle, and western blot was used to detect the expression of signal transduction and activator of transcription 3 (STAT3) pathway-related proteins in HepG2 cells treated with CSE. A nude mouse model with an orthotopic liver tumor was prepared. The mice were randomly divided into four groups, each containing 12 animals: the model group, the 5-FU group, the CSE10 group [10 mg/(kg·d)] and the CSE50 group [50 mg/(kg·d)]. The volume and mass changes in the nude mice with orthotopic transplanted tumors were observed. Western blot method was used to test the protein expression levels of p-STAT3 and p38 mitogen-activated protein kinase (p38MAPK). Tissues from the liver of mice in the model group and the CSE50 group were analyzed by using a protein tyrosine kinase (PTK) chip, and the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) function enrichment analysis of the differentially expressed proteins was performed.ResultsThis study showed that CSE significantly inhibited the proliferation of HepG2 cells (P < 0.05). After 48 h of CSE treatment, the cell cycle of HepG2 cells manifested as S phase and G2/M phase; p-STAT3 protein levels in the CSE groups were significantly lower than that in the control group (P < 0.05). Analysis of the tumor inhibition in the mice showed that the tumor masses and volume in CSE groups were lower (P < 0.05). The protein levels of p-STAT3 and p38MAPK in CSE50 group and 5-FU group decreased significantly (P < 0.05). PTK antibody chip screening results showed that CSE groups had a bidirectional regulation trend, and there were 23 up-regulated PTKs and six down-regulated PTKs. The GO and KEGG analyses showed that CSE exerted its anticancer effects through regulation of biological processes, including mitogen-activated protein kinase (MAPK) cascade, chemotaxis, cell invasion, cell adhesion, angiogenesis and other biological processes, and through signaling pathways, including the MAPK, phosphatidylinositol-3-kinase/serine threonine protein kinase (PI3K/AKT), and RAS signaling pathways.ConclusionsCSE can effectively inhibite the proliferation of HepG2 cells and effectively inhibite the growth of liver cancer orthotopic transplantation tumor. Its mechanism may be closely related to the regulation of STAT3, MAPK and PI3K/AKT signaling pathways.
Compound Chai Jin Jie Yu Tablets, Acts as An Antidepressant by Promoting Synaptic Function in the Hippocampal Neurons
LI Zi-Rong, HAN Yuan-Shan, WU Meng-Yao, LIU Jian, JIN Shi, ZHANG Xi, WANG Yu-Hong
2020, 3(2): 80-95. doi: 10.1016/j.dcmed.2020.06.003
[Abstract](77) [FullText HTML] (39) [PDF 1125KB](5)
ObjectiveTo investigate the effectiveness of compound Chai Jin Jie Yu Tablets (CJJYT) in ameliorating cognitive impairment associated with depression and its potential mechanism of action.MethodsIn vitro experiments, the hippocampus was isolated from the whole brain of the fetal rat and cultured into hippocampal neuron cells. 50 μM corticosterone (CORT) was added to each group 18 h before the experiment for modeling depression, with the exception of the control group. After modeling, the blank serum group was added with 10% blank serum, the CJJYT group and the venlafaxine group were added with the corresponding 10% drug-containing serum, and the control group and the model group were added with equal volumes of culture medium. The intracellular Ca2+ mean fluorescence intensity, miniature excitatory postsynaptic current (mEPSC) current amplitude, and frequency of different hippocampal neurons were evaluated as indicators of synaptic function in the hippocampal neurons. In addition, the expression of synaptic plasticity related proteins, synaptophysin-α (SYN-α), N-methyl-D-aspartate receptor 2A (NR2A), N-methyl-D-aspartate receptor 2B (NR2B), post synaptic density 95 protein (PSD-95), calcium/calmodulin dependent protein kinaseⅡ (CaMKⅡ) and synaptic associated protein (SynGAP) were detected in the hippocampal neurons by immunofluorescence staining and high content analysis (HCA) system. Then, reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expression levels of SYN-α, NR2A, NR2B, PSD-95, CaMKⅡ and SynGAP. For in vivo experiments, except for those in the blank control group, all rats were treated within a single cage for chronic unpredictable stress-induced depression modeling and subjected to corresponding drug interventions. Behavioral tests were used to detect depressive behavior and determine learning, memory and other cognitive abilities, whereas enzyme-linked immunosorbent assay (ELISA) was used to detect the CORT levels. Golgi-Cox staining was used to observe changes in the synaptic morphology of parahippocampal gyrus CA1 area (CA1) and dentategyrus (DG).ResultsIn vitro, CJJYT treatment reduced the intracellular Ca2+ mean flurorescence intensity in the hippocampal neurons (P < 0.05), causing a reduction in the frequency and current amplitude of mEPSC (P < 0.05), and thus inhibited the excessive activation of post-synaptic receptors. CJJYT treatment reduced the protein and mRNA expression of SYN-α, NR2A, NR2B and PSD-95 in the hippocampal neurons (P < 0.05), increased the mRNA and protein expression of CaMKⅡ and SynGAP (P < 0.05), and thereby improved the synaptic plasticity of the hippocampal neurons. In vivo, CJJYT intervention improved sucrose preference, voluntary activity , learning and memory ability of Morris water maze test, and suppressed appetite (P < 0.05), and increased the despair feeling of forced swimming test (P < 0.05). The CORT level was reduced (P < 0.05), leading to the repair of synaptic damage in the hippocampal neurons.ConclusionsCJJYT can improve the synaptic function of hippocampal neurons and has obvious protective effects on neurons. It can repair the structural damage in the hippocampal neurons, improving the cognitive ability of the depressed model rats. The mechanism of CJJYT improving cognition in depressed rats may be related to the transmission and function of SYN-α/NR and its downstream neurotransmitters.
Systematic Pharmacological Strategies to Explore the Regulatory Mechanism of Ma Xing Shi Gan Decoction on COVID-19
ZHANG Shi-Ying, LI Ling, ZHANG Ning, XIA Hong-Tao, LU Fang-Guo, LI Wei-Qing
2020, 3(2): 96-115. doi: 10.1016/j.dcmed.2020.06.004
[Abstract](95) [FullText HTML] (45) [PDF 1937KB](8)
ObjectiveTo use systematic pharmacological strategies to explore the regulatory mechanisms of Ma Xing Shi Gan Decoction (MXSGD) against the coronavirus disease 2019 (COVID-19).MethodsData on the compounds and targets of MXSGD were collected from the Traditional Chinese Medicene Systems Parmacology Database and Analysis Platform (TCMSP) and TCM Databases@Taiwan. Data on ACE2-related targets and the protein-protein interaction (PPI) were collected from the String database. The Cytoscape 3.7.2 was used to construct and analyze the networks. The DAVID platform was used for Gene Ontology (GO) and pathway enrichment analyses.ResultsData on 272 MXSGD targets and 21 SARS-CoV-2 potential targets were collected. Four networks were constructed and analyzed based on the data: (1) compound-target network of MXSGD; (2) MXSGD-SARS-CoV-2-PPI network; (3) cluster of MXSGD-SARS-CoV-2-PPI network; (4) Herb-Pathway-Target network. The core targets included AKT1, MAPK3, IL-6, TP53, VEGFA, TNF, CASP3, EGFR, EGF and MAPK1. The antiviral biological processes were inflammatory responses (inflammatory cells, inflammatory cytokines and their signaling pathways), immune responses (T cells, monocytes, B cells and other immune cells), immune factors (IFN-γ, TNF-α and so on), virus defense, humoral immunity and mucosal innate immune response. The antivirus-related signaling pathways included TNF, NOD-like receptor, FoxO, PI3K-AKT and Toll-like receptor signaling pathways.ConclusionsMXSGD can control disease progression by regulating multiple compounds and targets; it can reduce inflammation and balance immunity by regulating several proteins that interact with ACE2 and signaling pathways closely related to disease development.
Network Pharmacology Approach to Investigate the Preventive Mechanism of Hunan Expert Group Recommended Chinese Medicine Prevention No. 2 Prescription Against COVID-19
WU Hua-Ying, LI Jing, MAO Yi-Zhi, YUAN Zhi-Ying, LI Feng, LI Liang, HUANG Hui-Yong
2020, 3(2): 116-132. doi: 10.1016/j.dcmed.2020.06.005
[Abstract](51) [FullText HTML] (27) [PDF 2491KB](2)
ObjectiveTo explore the possible preventive mechanism of Hunan expert group recommended Chinese medicine prescription of No. 2 (Pre-No. 2) against coronavirus disease 2019 (COVID-19) by network pharmacology method.MethodsThe target proteins of effective components and active compounds in Pre-No. 2 were screened by searching the Tradi-tional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). A component-target-disease interac-tion network of Pre-No. 2 was constructed by Cytoscape 3.7.2, gene ontology (GO) analysis, and Kyoto encyclopedia of genes and genomes (KEGG) analysis of target protein pathway by DAVID.ResultsA total of 163 compounds and 278 target protein targets in Pre-No. 2 were collected from the TCMSP database. Kaempferol, wogonin, 7-methoxy-2-methyl isoflavone, formononetin, isorhamnetin, and licochalcone A were the most frequent targets in the regulatory network. GO enrichment analysis showed that Pre-No. 2 regulated response to virus, viral processes, humoral immune responses, defense responses to virus and viral entry into host cells. KEGG enrichment analysis showed that the formula regulated the NF-κB signaling pathway, B cell receptor signaling pathway, viral carcinogenesis, T cell signaling pathway and FcγR-mediated phagocytosis signaling pathway.ConclusionsPre-No. 2 may play a preventive role against COVID-19 through regulation of the Toll-like signaling, T cell signaling, B cell signaling and other signaling pathways. It may re-gulate the immune system to protect against anti-influenza virus.
SARS-CoV-2 Infected Cases with Untraceable Epidemic Origin: Its Values, Identification and TCM Treatment in Preventing Recurrence
LI Kai, PIERRE Sirois, DAI Ai-Guo, LIAO Duan-Fang
2020, 3(2): 133-138. doi: 10.1016/j.dcmed.2020.06.006
[Abstract](45) [FullText HTML] (26) [PDF 489KB](0)
During the outbreak of coronavirus disease 2019 (COVID-19), the identification of a SARS-CoV-2 infected case with untraceable epidemic origin has three values: (1) the region has community transmission of the virus; (2) a certain portion of population in the community is getting immunized and more individuals have already been immunized; (3) an unpredictable future risk exists for regions where there are no infected cases with untraceable epidemic origin. Minimizing or avoiding the aggregation infection through individuals with no clinical symptoms is crucial and possible as its occurrence is mainly attributed to the local environment as opposed to the super spreader with or without clinical symptoms. As infected cases are not necessarily positive with gene test by definition, proper application of gene test is crucial in the identification of asymptomatic cases. In the early stage of an outbreak of infectious disease, gene test can be used to identify asymptomatic cases but it should not be used to exclude cases with typical clinical symptoms and signs. In the middle or late stages of an outbreak gene test should be applied in evaluation of infection rate of the population of a region, in addition to be used in spreader identification and isolation. Although asymptomatic cases are attributed to the overwhelming effect of personalized defense against pathogens, they are still a source of infection. Asymptomatic cases are considered to be Qi deficiency and with lingering toxicity in traditional Chinese medicine (TCM). Treatment in such cases, typically, involves therapy focused on replenishing the Qi, tonifying the lung, clearing the fever, and detoxification in order to return to a normal health condition. The recommended Chinese medicines include Qing Fei Pai Du Decoction, Yi Qi Qu Du Decoction, Xuan Fei Bai Du Granule, Lian Hua Qing Wen Capsule (Granule), and Jin Hua Qing Gan Granule, etc.
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2020, 3(2): 139-140.
[Abstract](37) [FullText HTML] (23) [PDF 143KB](1)