Ajuforrestin A, an Abietane Diterpenoid from Ajuga ovalifolia var. calanthe, Induces A549 Cell Apoptosis by Targeting SHP2.

The Src-homology 2 domain-containing phosphatase 2 (SHP2), which is encoded by PTPN11, participates in many cellular signaling pathways and is closely related to various tumorigenesis. Inhibition of the abnormal activity of SHP2 by small molecules is an important part of cancer treatment. Here, three abietane diterpenoids, named compounds 1-3, were isolated from Ajuga ovalifolia var. calantha. Spectroscopic analysis was used to identify the exact structure of the compounds. The enzymatic kinetic experiment and the cellular thermal shift assay showed compound 2 selectively inhibited SHP2 activity in vitro. Molecular docking indicated compound 2 targeted the SHP2 catalytic domain. The predicted pharmacokinetic properties by SwissADME revealed that compound 2 passed the majority of the parameters of common drug discovery rules. Compound 2 restrained A549 proliferation (IC50 = 8.68 ± 0.96 µM), invasion and caused A549 cell apoptosis by inhibiting the SHP2-ERK/AKT signaling pathway. Finally, compound 2 (Ajuforrestin A) is a potent and efficacious SHP2 inhibitor and may be a promising compound for human lung epithelial cancer treatment.

Vitexin protects melanocytes from oxidative stress via activating MAPK-Nrf2/ARE pathway.

INTRODUCTION: Vitiligo is the most common type of depigmented skin disease. Cellular oxidative stress caused by reactive oxygen species (ROS) has been implicated in the pathogenesis of vitiligo. Nuclear factor E2-related factor 2 (Nrf2)/antioxidant response element (ARE) pathway plays an important role in melanocytes against hydrogen peroxide (H2O2) induced oxidative stress. In addition, vitexin may protect vitiligo by inhibiting oxidative stress and inflammation. OBJECTIVE: In the present study, we aimed to investigate the antioxidant effect of vitexin-activated mitogen-activated protein kinase (MAPK)-Nrf2/ARE axis in vitiligo. METHODS: MTT assay identified cell viability of human melanocyte PIG1. Cell apoptosis was evaluated by flow cytometry. Gene and protein expression levels were analyzed by quantitative real-time PCR (qPCR) and Western blotting. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expressions of inflammatory factors and ROS production. RESULTS: Vitexin inhibited H2O2-induced melanocyte apoptosis and promoted cell proliferation. Moreover, vitexin decreased expression of interleukin-1ß (IL-1ß), IL-17A, and ROS in melanocytes induced by H2O2. Subsequently, activation of MAPK-Nrf2/ARE signaling was readily induced by vitexin treatment, as evidenced by the upregulation of antioxidant genes including heme oxygenase 1 (HO-1) and superoxide dismutase (SOD). Knockdown of Nrf2 reversed the protective effect of vitexin on H2O2-induced melanocytes. And, knockdown of Nrf2 increased the expression of IL-1ß, IL-17A and ROS, and reduced HO-1 and SOD expression. CONCLUSIONS: Vitexin protected melanocytes from oxidative stress by activating MAPK-Nrf2/ARE signaling pathway. Our results suggested that the role of the Nrf2/ARE axis in the antioxidant defense of melanocytes, and the potential therapeutic strategy for vitiligo.

Psoriasis Vulgaris of Blood Heat Syndrome in Plasma Based on Widely Targeted Techniques.

Traditional Chinese medicine classifies psoriasis (Ps) according to clinical manifestations, and its different clinical manifestations imply the pathogenesis and material evolution basis of Ps, especially biomarkers that are meaningful to identification of Ps, treatment response, and elucidation of the pathogenesis of the disease. This study aims to screen differential metabolites in plasma of psoriasis vulgaris (PV) of blood heat syndrome based on a widely targeted metabolomic technique and to analyze syndrome metabolic markers and metabolic pathways. Forty-five PV patients were recruited, including 21 cases of the blood heat syndrome group (BH-PPG), 24 cases of the non-blood-heat syndrome group (NBH-PPG), and 30 healthy cases of the normal control group (NPG). The UPLC-MS/MS detection platform, a self-developed database, and multivariate statistical analysis were applied to investigate the plasma metabolic differences. The biomarkers related to blood heat syndrome were screened using the principal component analysis method. A total of 479 metabolites were detected in the three groups of plasma samples; 72 different metabolites were sorted out in the BH-PPG/NPG group, 82 in the NBH-PPG/NPG group, and 8 in the BH-PPG/NBH-PPG group. Differential metabolites mainly consist of metabolites of organic acids, amino acids, carbohydrates, and nucleotides. Multiple metabolites ginkgolic acid, pyrroloquinoline quinone, L-aspartic acid, and citramalic acid were expected to be the potential biomarkers of blood heat syndrome PV. The formation and evolution processes may be associated with disorders and regulation of metabolic pathways, ferroptosis, carbon metabolism, and purine metabolism.

Eriodictyol protects skin cells from UVA irradiation-induced photodamage by inhibition of the MAPK signaling pathway.

Solar UVA irradiation-generated reactive oxygen species (ROS) induces the expression of matrix metalloproteinase 1 (MMP-1), leading to photoaging, however the molecular mechanism remains unclear. In the present study, we found that eriodictyol remarkably reduces UVA-mediated ROS generation and protects the skin cells from oxidative damage and the ensuing cell death. Moreover eriodictyol pretreatment significantly down-regulates the UVA-induced MMP-1 expression, and lowers the inflammatory responses within the skin cells. Pretreatment with eriodictyol upregulates the expression of tissue inhibitory metalloproteinase 1 (TIMP-1) and collagen-I (COL-1) at the transcriptional level in a dose-dependent manner. UVA-induced phosphorylation levels of c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK) and p38 leading to increased MMP-1 expression are significantly reduced in eriodictyol-treated skin cells. In addition, eriodictyol pretreatment significantly suppresses inflammatory cytokines and inhibits the activation of MAPK signaling cascades in skin cells. Taken together, our results demonstrate that eriodictyol has both potent anti-inflammatory and anti-photoaging effects.

The association of serum vascular endothelial growth factor levels and psoriasis vulgaris: A protocol for systematic review and meta-analysis.

BACKGROUND: In recent years, more and more attention has been paid to the role of skin microcirculation in the pathogenesis of psoriasis. The vascular network of the skin is mainly distributed in the dermis and the subcutaneous fat layer join. The microvessels are composed of terminal arterioles, arteriovenous capillaries, and postcapillary venules. Vascular endothelial growth factor (VEGF) plays an important role in the pathogenesis of psoriasis by promoting angiogenesis. The purpose of this study is to evaluate the relationship between serum VEGF and psoriasis vulgaris. METHODS: Embase, CENTRAL, PubMed, China Biology Medicine Database, China National Knowledge Database, Wan Fang Database, and Chong Qing VIP Database will be searched to collect case-control studies and cohort studies and evaluate the relationship between serum VEGF and psoriasis vulgaris. The search time limits will be from the establishment of the database to December 2020. Two researchers will independently screen the studies, extract data, and evaluate the risk of bias of the studies. The Meta-analysis will be carried out with the RevMan5.3 software. The quality of all included studies will be evaluated by the Newcastle-Ottawa scale. RESULTS: This study will evaluate the relationship between serum VEGF and the pathogenesis of psoriasis vulgaris. CONCLUSION: This study will provide a theoretical basis for the pathogenesis of psoriasis vulgaris. OSF REGISTRATION NUMBER: DOI 10.17605/OSF.IO/6DV8P.

Identification of Pannexin 2 as a Novel Marker Correlating with Ferroptosis and Malignant Phenotypes of Prostate Cancer Cells.

PURPOSE: Prostate cancer (PCa) is a widespread urinary neoplasm and one of the most prevalent and second most frequent malignancies diagnosed in males worldwide. This study aimed to identify a candidate marker and explore its molecular mechanism in PCa. METHODS: Gene expression datasets, GSE55945 (n=21) and GSE46602 (n=50), were downloaded from the Gene Expression Omnibus database. Bioinformatic approaches were applied to identify potential markers. Effects of the candidate marker on proliferation, migration, invasion, and ferroptosis (ferrous iron and malondialdehyde (MDA)) in PCa cells and its mechanism were assessed after performing cell transfection. RESULTS: A total of 1435 common differentially expressed genes were identified in GSE55945 and GSE46602. Five key gene modules were listed based on a protein-protein interaction network, containing five hub genes. Pannexin 2 (PANX2), a candidate marker was identified, and findings revealed substantial upregulation of its expression levels in PCa cell lines. Blocking expression of PANX2 resulted in suppression of proliferation, migration, and invasion in PCa cells, while increasing ferrous iron and MDA levels. However, these effects were rescued by Nrf2 activator, oltipraz. The Nrf2 signaling pathway was consequently applied to determine underlying mechanism of PANX2 in PCa cells. We established that silencing PANX2 remarkably reduced protein expression levels in members of Nrf2 signaling pathway (Nrf2, HO-1, and FTH1). CONCLUSION: Our study demonstrated that PANX2 is implicated in the pathogenesis of PCa, which regulates malignant phenotypes and ferroptosis through Nrf2 signaling pathway, and maybe a potential therapeutic target for PCa.

miR124-3p/FGFR2 axis inhibits human keratinocyte proliferation and migration and improve the inflammatory microenvironment in psoriasis.

Keratinocyte hyperproliferation has been regarded as a central event in psoriasis pathogenesis. Investigating the mechanisms of keratinocyte hyperproliferation might provide novel strategies for psoriasis treatment. we demonstrated that fibroblast growth factor receptor 2 (FGFR2) expression was abnormally upregulated within psoriatic lesion tissues and HaCaT cells under rIL-22 stimulation. FGFR2 silence within HaCaT cells under rIL-22 stimulation significantly inhibited the capacity of cells to proliferate and to migrate, reduced IL-17A and TNFα mRNA expression, and decreased the protein levels of FGFR2, keratin 6, keratin 16, MMP1, MMP9, p-PI3K, p-AKT and p-ERK. In contrast to FGFR2, the expression of miR-124-3p showed to be remarkably downregulated within psoriasis lesion tissue samples and rIL-22-stimulated HaCaT cells. miR-124-3p inhibited the expression of FGFR2 via direct binding to its 3'UTR. Within HaCaT cells under rIL-22 stimulation, the overexpression of miR-124-3p also suppressed the capacity of cells to proliferate and to migrate, reduced IL-17A and TNFα mRNA expression, and decreased the protein levels of FGFR2, keratin 6, keratin 16, MMP1, MMP9 and p-PI3K, p-AKT and p-ERK. More importantly, when co-transfected to HaCaT cells, FGFR2-overexpressing vector significantly attenuated the effects of miR-124-3p mimics on HaCaT cells. In conclusion, we demonstrated an miR124-3p/FGFR2 axis that might inhibit human keratinocyte proliferation, migration, and improve the inflammatory microenvironment in psoriasis. miR124-3p/FGFR2 axis could be an underlying target for psoriasis therapy, which requires further in vivo and clinical investigation.

Long noncoding RNA TUG1 regulates prostate cancer cell proliferation, invasion and migration via the Nrf2 signaling axis.

BACKGROUND: Long noncoding RNAs (lncRNAs) have been identified to modulate the development and progression of prostate cancer (PCa) via the regulation of their target genes. However, the biological function underlying the effect of lncRNA TUG1 in PCa remains unclear. METHODS: Reverse transcription-quantitative polymerase chain reaction (qRT-PCR) and Western blotting analysis were used to assess the mRNA expression of TUG1 and protein expression levels of Nrf2 pathway members, respectively. The migration, invasion, and proliferation abilities of cells were assessed by the wound-healing, Transwell migration/invasion, and CCK8 assays, respectively. RESULTS: TUG1 was strikingly upregulated in PCa cells compared with non-tumorigenic human prostate epithelial cells. The LncTar Web Server, which is a bioinformatics tool, was used to predict the target association between TUG1 and Nrf2. Moreover, the expression of TUG1 showed a strikingly positive correlation with that of Nrf2 in TCGA PCa RNA-Seq data (r = 0.26,P = 4.63E-09). Subsequently, inhibition of TUG1 using siRNA resulted in deceased proliferation, migration, and invasion of PCa cells; however, these effects were reversed by treatment with oltipraz (an activator of Nrf2). Finally, we evaluated the Nrf2 pathway to reveal the underlying mechanism of TUG1 in PCa cells, and found that TUG1 knockdown decreased the protein expression of Nrf2 downstream members (e.g., HO-1, FTH1, and NQO1). CONCLUSIONS: LncRNA TUG1 plays an oncogenic role in human PCa cells by promoting the cell proliferation and invasion in PCa cell lines, at least partly via the Nrf2 signaling pathway.

miR-203 promotes HaCaT cell overproliferation through targeting LXR-α and PPAR-γ.

Psoriasis is an immune-mediated chronic inflammatory skin disease. Keratinocyte hyperproliferation has been regarded as a significant event in psoriasis pathogenesis. Considering the vital role of miRNA-mediated mRNA repression in psoriasis pathogenesis, in the present study, we attempted to investigate the mechanism of keratinocyte overproliferation from the point of miRNA-mRNA regulation. Both online microarray expression profiles and experimental results indicated that the expression of LXR-α and PPAR-γ was downregulated in psoriasis lesion skin. LXR-α or PPAR-γ overexpression alone was sufficient to inhibit keratinocyte proliferation, decrease KRT5 and KRT14 protein levels and increase KRT1 and KRT10 protein levels. miR-203 negatively regulated LXR-α and PPAR-γ expression through direct targeting. miR-203 inhibition exerted the opposite effects to LXR-α or PPAR-γ overexpression on HaCaT cells. More importantly, LXR-α or PPAR-γ overexpression could markedly remarkably attenuate the effects of miR-203 overexpression in keratinocytes, indicating that miR-203 promotes keratinocyte proliferation by targeting LXR-α and PPAR-γ. In conclusion, the miR-203-LXR-α/PPAR-γ axis modulates the proliferation of keratinocytes and might be a novel target for psoriasis treatment, which needs further in vivo investigation.

Ultrafast one-pot anodic preparation of Co3O4/nanoporous gold composite electrode as an efficient nonenzymatic amperometric sensor for glucose and hydrogen peroxide.

For fabrication of composite electrode, one-pot strategy is highly attractive for convenience and efficiency. Here, a self-supporting Co3O4/nanoporous gold (NPG) composite electrode was one-pot prepared via one-step in situ anodization of a smooth gold electrode in a CoCl2 solution within 100 s. It worked as a bifunctional electrocatalyst for glucose oxidation and H2O2 reduction in NaOH solution. Under optimized conditions, the electrocatalytic oxidation of glucose exhibits a wide linear range from 2 µM to 2.11 mM with a limit of detection as low as 0.085 µM (S/N = 3) and an ultrahigh sensitivity of 4470.4 µA mM-1 cm-2. Detection of glucose in human serum samples are also realized with results comparable to those from local hospital. The electrocatalytic reduction of H2O2 shows a linear response range from 20 µM to 19.1 mM and a high sensitivity of 1338.7 µA mM-1 cm-2. The present results demonstrate that the facilely prepared Co3O4/NPG is a promising nonenzymatic sensor for rapid amperometric detection of glucose and H2O2 with ultrasensitivity, high selectivity, satisfactory reproducibility, good stability and long duration.

Ultra-rapid fabrication of highly surface-roughened nanoporous gold film from AuSn alloy with improved performance for nonenzymatic glucose sensing.

Using one-step anodization strategy, a nanoporous gold film (HNPG) with large surface area was rapidly fabricated on Au80Sn20 (wt%) alloy in just 80 s. The formation of highly surface-roughened nanoporous structures results from a complex process of electrochemical dealloying of Sn component from AuSn alloy, anodic electrodissolution, disproportion and deposition of Au component, and spontaneous redox reaction between electrodissolved Sn2+ and AuCl4－species at the applied anodic potential. As-prepared HNPG/AuSn shows enhanced electrochemical performance for glucose oxidation in alkaline electrolyte. At a low potential of 0.1 V (vs. SCE), it offers a short response time of 4 s, a wide linear detection range of 2 µM to 8.11 mM, an ultralow detection limit of 0.36 µM (S/N = 3), an ultrahigh sensitivity of 4374.6 µA cm-2 mM-1, and satisfactory selectivity and reproducibility. Specifically, after 6 weeks, no obvious loss of glucose amperometric signal was observed on HNPG/AuSn. The facile preparation and excellent sensing performance of HNPG/AuSn electrode make sure that it is a promising candidate for advanced enzyme-free glucose sensors.

MiR-330 inhibits IL-22-induced keratinocyte proliferation through targeting CTNNB1.

Psoriasis is a common chronic inflammatory skin disease which is characterized by hyperproliferation and aberrant differentiation of keratinocytes; however the exact pathogenesis is largely unknown. Interleukin-22 (IL-22) has demonstrated its vital role in T cell-mediated immune response by interacting with keratinocytes in the pathogenesis of psoriasis. The microRNAs (miRNAs) are a class of small non-coding RNA molecules that play important roles in cellular processes by regulating gene expression at the post-transcriptional level. MiR-330 has been reported to inhibit the proliferation and migration of mouse keratinocytes. In the present study, we indicated that miR-330 expression in lesion tissue of psoriasis patients was specifically down-regulated, and could inhibit IL-22-induced proliferation of HaCaT and HKC cell. Wnt/ß-catenin pathway plays an essential role in the pathogenesis of psoriasis. By direct targeting CTNNB1, miR-330 could significantly downregulate IL-22-induced CTNNB1 expression. In addition, we found that the downstream targets of ß-catenin, CyclinD1 and Axin2, could be affected by miR-330; miR-330 could suppress CyclinD1 protein expression and rescue Axin2 protein expression. Taken together, we indicated miR-330 inhibits IL-22-induced proliferation of HaCaT and HKC cell by targeting CTNNB1 and subsequently affect the downstream factors, CyclinD1 and Axin2 for the first time, and provide diagnostic markers and a novel target for psoriasis treatment.

MicroRNA-146a and miR-99a are potential biomarkers for disease activity and clinical efficacy assessment in psoriasis patients treated with traditional Chinese medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Psoriasis is a common chronic inflammatory skin disease. A number of clinical investigations have indicated that traditional Chinese medicine (TCM) is an effective and safe treatment for psoriasis. Zhuhuang Granule (ZG) is a modified formulation of Zhuhuang Decoction, which is used traditionally in China for the treatment of psoriasis in clinical practice. AIM OF THE STUDY: Recent studies have found that microRNAs (miRNAs) play important roles in the pathogenesis of some skin diseases. The objective of our study was to investigate the effect of ZG on the expression of miRNAs in peripheral blood mononuclear cells (PBMCs) from psoriasis patients and to identify specific miRNA biomarkers for psoriasis disease activity and assessment of clinical efficacy. MATERIALS AND METHODS: Twenty-five psoriasis patients and 15 healthy control subjects were recruited to participate in this study from October 2013 to October 2014. Microarray and quantitative real-time PCR (qRT-PCR) were used to measure the global miRNA expression in PBMCs from psoriasis patients and healthy control subjects. We also measured the changes in the Psoriasis Area and Severity Index (PASI) score and miRNA expression of patients before and after treatment with ZG. RESULTS: The microarray results showed that 26 miRNAs were upregulated and 13 miRNAs were decreased in psoriasis patients. qRT-PCR validated 3 upregulated miRNAs (miR-146a, miR-31, miR-192-5p) and 2 downregulated miRNAs (miR-99a, miR-200c) in PBMCs from psoriasis patients compared with healthy controls (p<0.01). Moreover, after 8 weeks of ZG treatment, patients achieved a significant reduction in PASI scores. QRT-PCR analysis indicated that the expression of miR-146a and miR-99a is closely correlated with psoriasis severity (R2=0.772, p<0.01; R2=0.672, p<0.01). CONCLUSION: We suggest that both miR-146a and miR-99a may serve as potential biomarkers for disease activity and clinical efficacy in psoriasis patients treated with ZG.