Antiviral effect of baicalein on Porcine Deltacoronavirus infection by regulating the inflammatory responses through PI3K-Akt-NF-κB signaling pathway in cultured cells.

Porcine deltacoronavirus (PDCoV) is an newly emerged enteropathogenic coronavirus, mainly causing diarrhea in suckling piglets, and also has the potential for cross-species transmission. However, there are no effective vaccines or specific therapeutic agents for PDCoV. This study investigates the antiviral properties of baicalein against PDCoV infection in swine testicle cells (ST). It reveals that baicalein exerts a dose-dependent inhibitory effect on PDCoV replication, primarily targeting the replication stage of the viral infection by impeding viral RNA and protein synthesis. Furthermore, treatment with baicalein leads to reduced phosphorylation of PI3K, AKT, and NF-κB p65 proteins, along with decreased mRNA levels of pro-inflammatory cytokines (IL-1ß, IL-6, IL-8, and TNF-α). These results signify that PDCoV replication is inhibited through the inhibition of the PI3K-Akt-NF-κB protein signaling pathway, thereby suppressing the inflammatory response. In conclusion, it underscores the potential of baicalein as a therapeutic candidate for treating PDCoV infection.

Benzoylaconitine Alleviates Progression of Psoriasis via Suppressing STAT3 Phosphorylation in Keratinocytes.

Psoriasis is a chronic and multifactorial skin disease which is caused by inflammatory infiltrates, keratinocyte hyperproliferation, and accumulation of immune cells. As part of the Aconitum species, Benzoylaconitine (BAC) shows potential antiviral, anti-tumor, and anti-inflammatory effects. In this study, we investigated the effects and mechanisms of BAC on tumor necrosis factor-alpha (TNF-α)/LPS-induced HaCaT keratinocytes in a imiquimod(IMQ)-induced mice model. The results showed that BAC could relieve the symptoms of psoriasis by inhibiting cell proliferation, the release of inflammatory factors, and the accumulation of Th17 cells, while no obvious effect on cell viability and safety was observed both in vitro and in vivo. Additionally, BAC can markedly inhibit the protein and mRNA levels of inflammatory cytokines in TNF-α/LPS-induced HaCaT keratinocytes by inhibiting the phosphorylation of STAT3. In brief, our data indicated that BAC could alleviate the progression of psoriasis and may be a potential therapeutic agent for treating psoriasis in clinical practice.

Nuclear localization signal peptides enhance genetic transformation of Dunaliella salina.

BACKGROUND: Dunaliella salina (D. salina) expression system shows a very attractive application prospect, but it currently has a technical bottleneck, namely the low or unstable expression of recombinant proteins. Given the characteristics of cell-penetrating peptides or/and nuclear localization signal (NLS) peptides, this study is the first attempt to improve the transformation rate of foreign gene with trans-activating transcriptional (TAT) protein or/and NLS peptides. METHODS AND RESULTS: Using salt gradient method, exogenous plasmids were transferred into D. salina cells with TAT or TAT/NLS complexes simultaneously. The ß-glucuronidase gene expression was identified by means of histochemical stain and RT-qPCR detection. Through observation with light microscope, TAT-mediating cells exhibit an apparent cytotoxicity even at ratios of 0.5, no significant toxicity was noted in the TAT/plasmid/NLS complex group. It is obvious that with the addition of peptides the toxicity decreases significantly. Histochemical staining showed that the transformants presented blue color under light microscope, but the negative control and blank control are not. Furthermore, based on a TAT/plasmids ratio of 4 with 10 µg NLS peptides mediation, RT-qPCR results demonstrated that the transcripts of target gene were increased by 269 times than that of control group. CONCLUSIONS: This study demonstrated that combination of TAT and NLS peptides can significantly improve the transformation rate and expression level of foreign gene in D. salina system. It offers a promising way for promoting the application and development of D. salina bioreactor.

Osthole Induces Apoptosis and Inhibits Proliferation, Invasion, and Migration of Human Cervical Carcinoma HeLa Cells.

PURPOSE: To study the effect of osthole extract on proliferation, migration, invasion, and apoptosis of human cervical carcinoma HeLa cells and investigate its underlying mechanism. METHODS: HeLa cells were exposed to osthole at various concentrations. Cell viability, migration, and invasion were detected by MTT assay, scratch wound-healing assay, and invasion assay, respectively. The proportion of cells undergoing apoptosis was analyzed by flow cytometry. Western blot and RT-qPCR were performed to determine changes in the expression of key factors in the Wnt/ß-catenin signaling pathway. RESULTS: The osthole extract effectively inhibited the proliferation, migration, and invasion potential of HeLa cells in a dose-dependent manner. The rate of apoptosis induction in HeLa cells treated with the osthole extract for 48 h was significantly higher than that of the untreated controls. Outcomes of the western blotting analysis and RT-qPCR showed that the expression of ß-catenin, c-Myc, cyclin D1, survivin, and MMP-9 was significantly inhibited. CONCLUSION: Osthole could significantly inhibit the malignant behavior of HeLa cells and induce cellular apoptosis. Inactivation of the Wnt/ß-catenin signaling pathway by osthole may be a mechanism to control cancer metastasis.

S-palmitoylation of swine interferon-inducible transmembrane protein is essential for its anti-JEV activity.

Japanese encephalitis virus (JEV) is an infectious pathogen spreading in a wide range of vertebrate species. Pigs are amplifying hosts of JEV and thought to be maintained in nature predominantly by avian-mosquito cycles. In the innate immune system, interferon-inducible transmembrane protein (IFITM) is a small transmembrane protein family and has been identified as the first line of defense against a broad range of RNA virus invasion. In this paper, we found that swine IFITM (sIFITM) could restrict the replication of both JEV vaccine strain and wild strain NJ-2008. The cysteine S-palmitoylation modification of sIFITM plays important roles in their anti-JEV effects and intracellular distributions. Our findings show the anti-JEV activities of swine interferon-inducible transmembrane proteins and broaden the antiviral spectrum of IFITM protein family. The preliminary exploration of S-palmitoylation modification of sIFITM may contribute to understanding of the antiviral molecular mechanism of sIFITM.

Three Tiaobu Feishen formulae reduces cigarette smoke-induced inflammation in human airway epithelial cells.

OBJECTIVE: To investigate the therapeutic efficacy of Tiaobu Feishen formulae (TBFS) on cigarette smoke-induced inflammation in vitro using lipopolysaccharide (LPS)-induced and cigarette smoke extract (CSE)-induced NCI-H292 cells. METHODS: We evaluated the inhibitory effects of Bufei Jianpi formula (BJF), Bufei Yishen formula (BYF), and Yiqi Zishen formula (YZF) on the expressions of inflammatory cytokines including tumor necrosis factor (TNF)-α and interleukin (IL)-8, matrix metalloproteinase (MMP)-9, tissue inhibitor of matrix metalloprotease (TIMP)-1, and superoxide dismutase (SOD) in H292 cells stimulated with LPS or CSE. Their related transcription factors and signaling pathways were also analyzed. RESULTS: BJF, BYF, and YZF significantly inhibited the LPS- or CSE-induced expressions of TNF-α, IL-8, MMP-9, TIMP-1, and SOD in H292 cells, and suppressed the activation of transcription factors including nuclear transcription factor (NF)-κB, activator protein (AP)-1, and signal transducers and activators of transcription (STAT) 3 and their corresponding pathways, including NF-κB, mitogen-activated protein kinase (MAPK), STAT3, and peroxisome proliferator-activated receptor (PPAR). CONCLUSION: BJF, BYF, and YZF effectively suppressed inflammatory responses, protease-antiprotease imbalance, and oxidative stress induced by LPS and CSE, an effect that was closely associated with the inhibition of the NF-κB, MAPK, STAT3, and PPAR pathways.

Three Tiaobu Feishen therapies protect human alveolar epithelial cells against cigarette smoking and tumor necrosis factor--induced inflammation by nuclear factor-kappa B pathway.

OBJECTIVE: To investigate the efficacy of Tiaobu Feishen formulae (TBFS), including Bufei Jianpi formula (BJF), Bufei Yishen formula (BYF), and Yiqi Zishen formula (YZF), on inflammatory response, protease-anti-protease imbalance and collagen deposition in rats. METHODS: In present work, we used an in vitro model of cigarette smoking extract (CSE)- and tumor necrosis factor-α (TNF-α)-induced A549 cells to examine the efficacy of BJF, BYF and YZF on the production of inflammatory cytokines, including TNF-α and interleukin (IL)-8, IL-6, matrix metalloproteinases (MMP)-9, and IL-10 in CSE or TNF-α-induced A549 cells. And their related transcription factors and signaling pathway were also analyzed. RESULTS: The results showed that BJF, BYF and YZF could significantly decrease the expression levels of the pro-inflammatory cytokines induced by CSE or TNF-α. Furthermore, BJF, BYF and YZF could suppress CSE- or TNF-α-induced activation of nuclear factor-kappa B (NF-κB) transcription factors and its corresponding pathways. Taken together, these data implied that BJF, BYF and YZF effectively inhibited CSE- or TNF-α-induced inflammatory response in alveolar epithelial cell, which was due to their inhibition effect on NF-κB pathways. CONCLUSION: Our findings suggest that the Tiaobu Feishen therapies may protect human alveolar epithelial cells against cigarette smoking and TNF-α-induced inflammation. NF-κB pathway may involve in the actions.

LPS­induced proinflammatory cytokine expression in human airway epithelial cells and macrophages via NF­κB, STAT3 or AP­1 activation.

Lipopolysaccharide (LPS), the major outer surface membrane component of Gram-negative bacteria, is one of the main etiological factors in the pathogenesis of several lung diseases, such as chronic obstructive pulmonary disease. The respiratory epithelium and the macrophages comprise the dynamic interface between the outside environment and the host response to bacterial infection via cytokine secretion. In the present study, the mechanisms of LPS induced­inflammatory response in human lung cells and macrophages were investigated. The effects of LPS exposure on cytokine production, inflammation­related transcription factors and intracellular signaling pathway activation were assessed in human lung mucoepidermoid carcinoma H292 cells and human macrophage THP­1 cells. The results demonstrated that LPS markedly increased the expression of interleukin (IL)­6, IL­8, tumor necrosis factor (TNF)­α, matrix metallopeptidase (MMP)­9 and tissue inhibitor of metalloproteinases­1 in H292 cells, while it increased the production of IL­6, IL­8 and TNF­α in differentiated THP­1 cells. In addition, LPS exposure activated nuclear factor (NF)­κB and activator protein (AP)­1 signaling in H292 cells, while it activated NF­κB and signal transducer and activator of transcription (STAT) 3 signaling in THP­1 cells. Furthermore, treatment with NF­κB, AP­1 or STAT3 inhibitors significantly decreased the LPS­mediated expression of IL­8 and TNF­α in these cells, suggesting that these pathways might serve crucial roles in LPS­induced cytokine expression. In conclusion, LPS stimulation of H292 and THP­1 cells induced cytokine expression and NF­κB, mitogen­activated protein kinase and Janus kinase/STAT3 pathway activation with subsequent nuclear translocation of NF­κB, AP­1 and STAT3, which demonstrated potential of the use of NF­κB, AP­1 and STAT3 in therapies for conditions and diseases associated with chronic inflammation.

[Effect of Qige Powder on Angiogenesis Induced by Esophageal Cancer Cell Line Eca-9706].

OBJECTIVE: To study the effect Qige powder on esophageal cancer angiogenesis. METHODS: Inhibitive effect of Qige powder ethanol extract on proliferation of Esophageal cancer cell line Eca-9706 was detected by MTT assay. The Eca-9706 cells conditioned medium in Qige powder IC50 concentration were collected. Angiogenesis, as well as proliferation, migration and tube formation of human umbilical vein endothelial cells were observed by Chick embryo chorioallantoic membrane model(CAM), MTT assay, the migration and tube formation assay. VEGF and IL-6 contents in culture medium supernatant of Eca-9706 cells and human umbilical vein endothelial cells were detected by ELISA. RESULTS: Qige powder ethanol extract could inhibit the proliferation of Eca-9706 cells, with a certain dose-effect relationship, IC50 value was 96 µg/mL. Eca-9706 cells conditioned medium could significantly increase the CAM generating total vessel area, human umbilical vein endothelial cells proliferation,migration and tube formation, while the Eca-9706 cell conditioned medium of Qige powder ethanol extract could reduce CAM angiogenesis, human umbilical vein endothelial cells proliferation and tube formation, but increase endothelial cells migration. Qige powder ethanol extract could reduce endothelial cells secreting VEGF and IL-6 while increase Eca-9706 cells secreting. CONCLUSION: Qige powder ethanol extract can inhibit angiogenesis, endothelial cell proliferation, and tube formation induced by Eca-9706 cells, while reduce VEGF and IL-6 secretion of endothelial cells.

[Discussion of Chinese medical research ideas from miRNAs features].

In this paper, miRNAs features were briefly introduced and agreeable points were discussed from 4 aspects: organs relationship, syndrome research, Chinese medical pathogeneses, and actions of Chinese herbs. miRNAs, as information media for organs interrelation, was believed to explain Chinese medical pathogeneses and reveal partial molecular mechanisms of Chinese medicine. miRNAs in the body fluid could be taken as one of biological bases of syndromes.

[The relationship between inhibitory effect of xuanfuguilian prescription on the growth of esophageal cells ECA9706 and its components].

OBJECTIVE: To discuss the relationship between inhibitory effect of different concentration ethanol extracts from Xuanfuguilian prescription on the growth of esophageal cells ECA9706 and its components. METHODS: The drug was extracted with anhydrous ethanol, 95%, 85%, 75%, 50%, 25% ethanol or water, the cell proliferation inhibitory rate of different solvent extracts were detected with MTT assay, and IC50 was calculated. The components of the drug were determined by LC/MS. Based on the inhibitory rate and the components peak area, the samples were hierarchy clustered respectively. The correlation of components peak area and inhibition rate was analyzed with Pearson Correlation. The components peak area and inhibition rate were analyzed using PLS. RESULTS: The IC50 of the 7 extracts on esophageal carcinoma cell ECA9706 were respectively 46.361, 52.67, 58.11, 78.26, 93.10, 2579.43 and 3953.34 microg/ mL 22 stable peaks were determined by LC-MS. Based on the inhibition rate and the components peak area, the clustering results of the two samples were similar. The 10 peaks areaes were closely related to inhibition rate (P < 0.05) and the PLS between components peaks area and inhibition rate was constructed. CONCLUSION: Extracts with different concentration ethanol have different effects, and their curative effects are closely related to components.