Search

Background Wound healing is a complex physiological process that is crucial for reestablishing the epithelial barrier following injury. Objective The aim of this study was to demonstrate the efficacy of calcipotriol, a synthetic vitamin D3 analogue, in wound healing in an acute mice wound model. Methods An excision wound model was established in mice, and the wound healing activity of calcipotriol was evaluated. Human keratinocyte cell lines, HaCaT and NHEK, were utilized in in vitro skin wound healing model. Cytokine expression levels were measured by real-time PCR and ELISA assay. The expression of epithelial-mesenchymal transition (EMT)-associated molecules and the phosphorylation of Yes-associated protein (YAP) was determined by western blotting. Results The increase in re-epithelialization by calcipotriol treatment early in the wound was associated with the EMT process. A scratch assay using HaCaT and NHEK cells also showed that calcipotriol administration resulted in effective wound closure. We demonstrated that calcipotriol promoted keratinocyte migration by interfering with the Hippo pathway. Calcipotriol-mediated enhancement of cell migration is related to downregulated phosphorylation of YAP and increased levels of YAP and PDZ-binding motif (TAZ). Mechanistically, we defined that calcipotriol facilitated the crosstalk between the YAP/TAZ and TGF-β/Smad signaling pathways, eliciting EMT in keratinocytes during the wound healing process. Conclusions These results suggest that the positive effect of calcipotriol on keratinocyte migration is mediated by the induction of EMT via the regulation of Hippo pathway, which promotes the acceleration of wound closure.

More and more reports have pointed out that rotenone, as an insecticide, has high neurotoxicity and reproductive toxicity to livestock and mammals. As a highly physiological correlation system of internal organs, quasi-organs have great potential in the fields of drug toxicity and efficacy test, toxicology research, developmental biology and so on. In this study, brain organs (mBOs) derived from mouse neural stem cells were used to investigate the effects of rotenone on the physiological activity and epigenetic modification of mBOs. At the same time, Rotenone could significantly stimulate the increase of the concentration of LPO, lactic acid and hydroxyl radical in mBOs, and inhibit the expression of neuronal marker Tuj1, CHAT, PAX6 and so on. Further analysis showed that Rotenonem could induce mitochondrial damage in mBOs. The results of qPCR and Western blot showed that Rotenone could up-regulate the expressions of ferroptosis promoting protein p53, Cox2 and so on, while inhibit the expressions of negative regulatory protein of ferroptosis GPX4, FTH1, SLC7A11. In addition, the results of RRBS-Seq sequencing showed that the methylation modification at DMR level in Rotenone-treated mBOs group was significantly higher than that in Ctrl group. The results of KEGG analysis showed that compared with Ctrl, the genes with hypermethylation of promoter and Genebody in Rotenone-treated mBOs were mainly located in the Neuro active ligand-receptor interaction signal transduction pathway. In summary, rotenone can significantly lead to abnormal methylation of mouse brain organs, and lead to the loss of normal physiological function of neurons by inducing ferroptosis.

Human amniotic fluid stem cell-derived exosome (HuAFSC-exosome) transplantation is considered a promising treatment for premature ovarian failure (POF). However, its mechanism remains unclear. In this study, exosomes were isolated and enriched from HuAFSC subsets of CD44+/CD105+, and the exosomes were transplanted into a POF model in vitro and in vivo. Our results confirmed that the exosomes produced by CD44+/CD105+ HuAFSCs could achieve therapeutic effects in a mouse POF model. Our research also showed that CD44+/CD105+ HuAFSC-exosomes carrying miR-369-3p could specifically downregulate the expression of YAF2, inhibit the stability of PDCD5/p53, and reduce the apoptosis of ovarian granulosa cells (OGCs), thereby exerting therapeutic effects on POF. Knowledge of these mechanisms demonstrates that miRNAs carried by CD44+/CD105+ HuAFSC-exosomes are critical to the therapy of POF. This will be useful for the clinical application of stem cells.

Trimethylamine N-oxide (TMAO) leads to the development of cardiovascular and chronic kidney diseases, but there are currently no potent drugs that inhibit the production or toxicity of TMAO. In this study, high-fat diet-fed ApoE-/- mice were treated with finasteride, ranitidine, and andrioe. Subsequently, the distribution and quantity of gut microbiota in the faeces of the mice in each group were analysed using 16S rRNA sequencing of the V3+V4 regions. Pathological examination confirmed that both ranitidine and finasteride reduced atherosclerosis and renal damage in mice. HPLC analysis also indicated that ranitidine and finasteride significantly reduced the synthesis of TMAO and the TMAO precursor delta-Valerobetaine in their livers. The 16S rRNA sequencing showed that all 3 drugs significantly increased the richness and diversity of gut microbiota in the model mice. Bioinformatic analysis revealed that the faeces of mice treated with ranitidine and finasteride, had significant increases in the number of microbes in the families g_Helicobacter, f_Desulfovibrionaceae, Mucispirillum_schaedleri_ASF457, and g_Blautia, whereas the relative abundances of microbes in the families Enterobacter_sp._IPC1-8 and g_Bacteroides were significantly reduced. The microbiota metabolic pathways, such as nucleotide and cofactor and vitamin metabolism were also significantly increased, whereas the activities of metabolic signalling pathways related to glycan biosynthesis and metabolism and cardiovascular diseases were significantly reduced. Therefore, our study indicates that in addition to their known pharmacological effects, ranitidine and finasteride also exhibit potential cardiovascular and renal protective effects. They inhibit the synthesis and metabolism of TMAO and delay the deposition of lipids and endotoxins through improving the composition of the gut microbiota.

Objective To examine the role of high‐fat and high‐sugar (HFHS) diet‐induced oxidative stress, which is a risk factor for various diseases, in premature ovarian failure (POF). Materials and methods Ovarian granulosa cells (OGCs) were isolated from mice and cultured in medium supplemented with HFHS and poly (lactic‐co‐glycolic acid) (PLGA)‐cross‐linked miR‐146b‐5p nanoparticles (miR‐146@PLGA). RNA and protein expression levels were examined using quantitative real‐time polymerase chain reaction and Western blotting, respectively. HFHS diet‐induced POF model mice were administered miR‐146@PLGA. Results The ovarian tissue of mice fed a HFHS diet exhibited the typical pathological characteristics of POF. HFHS supplementation induced oxidative stress injury in the mouse OGCs, activation of the Dab2ip/Ask1/p38‐Mapk signalling pathway and phosphorylation of γH2A.X in vitro and in vivo. The results of the luciferase reporter assay revealed that miR‐146 specifically downregulated p38‐Mapk14 expression. Meanwhile, co‐immunoprecipitation and Western blot analyses revealed that HFHS supplementation upregulated nuclear p38‐Mapk14 expression and consequently enhanced γH2A.X (Ser139) phosphorylation. The HFHS diet‐induced POF mouse model treated with miR‐146@PLGA exhibited downregulated p38‐Mapk14 expression in the OGCs, mitigated OGC ageing and alleviated the symptoms of POF. Conclusions This study demonstrated that HFHS supplementation activates the Dab2ip/Ask1/p38‐Mapk signalling pathway and promotes γH2A.X phosphorylation by inhibiting the expression of endogenous miR‐146b‐5p, which results in OGC ageing and POF development., To examine the role of high‐fat and high‐sugar (HFHS) diet‐induced oxidative stress, which is a risk factor for various diseases, in premature ovarian failure (POF). Ovarian granulosa cells (OGCs) were isolated from mice and cultured in medium supplemented with HFHS and poly (lactic‐co‐glycolic acid) (PLGA)‐cross‐linked miR‐146 nanoparticles (miR‐146@PLGA). RNA and protein expression levels were examined using quantitative real‐time polymerase chain reaction and Western blotting, respectively. HFHS diet‐induced POF model mice were administered miR‐146@PLGA. The ovarian tissue of mice fed a HFHS diet exhibited the typical pathological characteristics of POF. HFHS supplementation induced oxidative stress injury in the mouse OGCs, activation of the Dab2ip/Ask1/p38‐Mapk signalling pathway and phosphorylation of γH2A.X in vitro and in vivo. The results of the luciferase reporter assay revealed that miR‐146 specifically downregulated p38‐Mapk14 expression. Meanwhile, co‐immunoprecipitation and Western blot analyses revealed that HFHS supplementation upregulated nuclear p38‐Mapk14 expression and consequently enhanced γH2A.X (Ser139) phosphorylation. The HFHS diet‐induced POF mouse model treated with miR‐146@PLGA exhibited downregulated p38‐Mapk14 expression in the OGCs, mitigated OGC aging and alleviated the symptoms of POF. This study demonstrated that HFHS supplementation activates the Dab2ip/Ask1/p38‐Mapk signalling pathway and promotes γH2A.X phosphorylation by inhibiting the expression of endogenous miR‐146, which results in OGC aging and POF development.

OBJECTIVE: To investigate the effects of total glucosides of paeony (TGP) on the proliferation and activation-induced cell death of mouse T cells and the mechanism underlying the immunosuppressive effects of TGP. METHODS: Purified total T cells isolated from the spleen of C57BL/6 mice were treated with TGP at 0, 50, 100, or 200 μg/mL and stimulated by anti-CD3/ CD28. Flow cytometry was performed to detect the cell death and the proliferation of CFSE-labeled T cells. The expression of Fas/FasL mRNA was detected using RT-PCR, and flow cytometry was used to analyze the expression of Fas/FasL proteins on activated T cells. Western blotting was used to detect the expression of Bcl-2 in the cells. RESULTS: TGP treatment for 48 h significantly reduced the total number and percentage of viable T cells and dose-dependently lowered the percentage of divided T cells. TGP treatment obviously up-regulated the cellular expression of Fas mRNA, enhanced Fas expression on the surface of the T cells, and decreased the expression level of Bcl-2 protein in the cells. CONCLUSION: TGP significantly inhibits proliferation and promotes activation-induced cell death of mouse T cell by increasing the expression of Fas and downregulating the expression of Bcl-2.

Background Huang Lian Jie Du Decoction (HLJDD), a very famous traditional Chinese medicinal prescription, has been used for heat dissipation and detoxification in China. This study was aimed to evaluate the reno-protective effects of HLJDD against lupus nephritis (LN) in vivo in MRL/lpr mice. Methods Animals were administered orally every day for eight consecutive weeks except the mice of normal group and model group. Organ indexes, serum interleukin-6 (IL-6), interleukin-10 (IL-10), interferon-gamma (IFN-γ) and the anti-double stranded DNA (anti-dsDNA) antibody were tested, respectively. Creatinine (Cr), blood urea nitrogen (BUN) and urine protein were measured for renal function evaluation. The expression of phosphorylated signal transducer and activator of transcription 3 (p-STAT 3) in kidney tissue was observed by western blot (WB) and immunohistochemical (IHC) method. Meanwhile, histopathological changes in the renal were studied by hematoxylin-eosin (H&E) staining. Results The mice of HLJDD-treated group exhibited a significant reduced mortality (p

Background The effects of short-term high fat diets on physiology are elusive and the molecular changes following fat overconsumption remain largely unknown. In this study, we aimed to evaluate exercise capacity in mice fed with a high fat diet (HFD) for 3 days and investigate the molecular mechanisms in the early response to high-fat feeding. Methods Exercise capacity was assessed by weight-loaded swimming test in mice fed a control diet (10 kcal% fat) or a HFD (60 kcal% fat) for 3 days. Global gene expression of ten important tissues (brain, heart, liver, spleen, lung, kidney, stomach, duodenum, skeletal muscle and blood) was analyzed using RNA Sequencing. Results A HFD for just 3 days can induce 71% decrease of exercise performance prior to substantial weight gain (P