Forsythoside A regulates pulmonary fibrosis by inhibiting endothelial-to-mesenchymal transition and lung fibroblast proliferation via the PTPRB signaling.

BACKGROUND: Pulmonary fibrosis (PF) is an end-stage change in many interstitial lung diseases, whereas no proven effective anti-pulmonary fibrotic treatments. Forsythoside A (FA) derived from Forsythia suspensa (Thunb.) Vahl, has been found to possess lung-protective effect. However, studies on its anti-pulmonary fibrosis effect are limited and its mechanism of action remains unknown. PURPOSE: This study aimed to explore the underlying mechanism of FA on PF. METHODS: Male C57BL/6 mice were randomized into normal (CON), model (BLM), pirfenidone (PFD), low- and high-dose FA (FA-L, FA-H, respectively). Except for the CON group, which was injected with the same dose of saline, the model of PF was established by intratracheal instillation of BLM, during which the survival rate and body weight changes of the mice were measured. The lung histopathology was evaluated by Hematoxylin-eosin, Sirius red, and Masson staining. Transcriptome analysis was performed to screen for the differential genes associated with the role of FA in PF. Differential genes in normal and pulmonary fibrosis patients with the GSE2052 dataset were analyzed in the GEO database. The levels of CTGF, α-SMA, MMP-8 in lung and TNF-α in bronchoalveolar lavage fluid (BALF) were detected by ELISA. The levels of HYP in lungs were detected by digestion. The mRNA and protein levels of MMP-7, E-cadherin, CD31, α-SMA, TGF-ß1, IL-6, ß-catenin, ZO-1, PTPRB, E-cadherin, and vimentin in lungs were detected by RT-qPCR and Western blot. The expression of CD31, α-SMA, TGF-ß1 and ZO-1 were detected by immunofluorescence. TGF-ß1-stimulated HFL1 cells and human umbilical vein endothelial cells (HUVECs) were used in an attempt to explore the possible role of protein tyrosine phosphatase receptor type B (PTPRB) involved in FA-induced improvement of PF. RESULTS: The results showed that FA could improve the survival rate and body weight of PF mice. FA could alleviate the symptoms of alveolar wall thickening, inflammatory cell infiltration, blue collagen fiber deposition, collagen fiber type Ⅰ and type Ⅲ in mice with PF. In addition, FA could reduce the levels of HYP, CTGF, α-SMA, TGF-ß1, TNF-α, ß-catenin and MMP8, and regulate the expression levels of CD31, ZO-1, PTPRB and E-cadherin in lung of mice with PF, inhibiting endothelial-to-mesenchymal transition (EndMT) and fibroblasts proliferation. In the GSE2052 dataset, the expression level of PTPRB is reduced in lung tissue from PF patients, and results from transcriptome sequencing indicate that PTPRB expression is also reduced in PF mice. In addition, the effect of FA on TGF-ß1-induced HFL1 or HUVECs cells could be attenuated by the inhibitor of PTPRB, suggesting that the effect of FA on PF is related to PTPRB. CONCLUSION: This study demonstrated that FA could ameliorate PF by inhibiting lung fibroblast proliferation and EndMT, and that PTPRB might be a target of FA to ameliorate PF, which provided evidence to support FA as a candidate phytochemical for PF.

Hierarchically Assembled Nanofiber Scaffolds with Dual Growth Factor Gradients Promote Skin Wound Healing Through Rapid Cell Recruitment.

To address current challenges in effectively treating large skin defects caused by trauma in clinical medicine, the fabrication, and evaluation of a novel radially aligned nanofiber scaffold (RAS) with dual growth factor gradients is presented. These aligned nanofibers and the scaffold's spatial design provide many all-around "highways" for cell migration from the edge of the wound to the center area. Besides, the chemotaxis induced by two growth factor gradients further promotes cell migration. Incorporating epidermal growth factor (EGF) aids in the proliferation and differentiation of basal layer cells in the epidermis, augmenting the scaffold's ability to promote epidermal regeneration. Concurrently, the scaffold-bound vascular endothelial growth factor (VEGF) recruits vascular endothelial cells at the wound's center, resulting in angiogenesis and improving blood supply and nutrient delivery, which is critical for granulation tissue regeneration. The RAS+EGF+VEGF group demonstrates superior performance in wound immune regulation, wound closure, hair follicle regeneration, and ECM deposition and remodeling compared to other groups. This study highlights the promising potential of hierarchically assembled nanofiber scaffolds with dual growth factor gradients for wound repair and tissue regeneration applications.

Anti-Inflammatory Peptide-Conjugated Silk Fibroin/Cryogel Hybrid Dual Fiber Scaffold with Hierarchical Structure Promotes Healing of Chronic Wounds.

Chronic wounds resulting from diabetes, pressure, radiation therapy, and other factors continue to pose significant challenges in wound healing. To address this, this study introduces a novel hybrid fibroin fibrous scaffold (FFS) comprising randomly arranged fibroin fibers and vertically aligned cryogel fibers (CFs). The fibroin scaffold is efficiently degummed at room temperature and simultaneously formed a porous structure. The aligned CFs are produced via directional freeze-drying, achieved by controlling solution concentration and freezing polymerization temperature. The incorporation of aligned CFs into the expanded fibroin fiber scaffold leads to enhanced cell infiltration both in vitro and in vivo, further elevating the hybrid scaffold's tissue compatibility. The anti-inflammatory peptide 1 (AP-1) is also conjugated to the hybrid fibrous scaffold, effectively transforming the inflammatory status of chronic wounds from pro-inflammatory to pro-reparative. Consequently, the FFS-AP1+CF group demonstrates superior granulation tissue formation, angiogenesis, collagen deposition, and re-epithelialization during the proliferative phase compared to the commercial product PELNAC. Moreover, the FFS-AP1+CF group displays epidermis thickness, number of regenerated hair follicles, and collagen density closer to normal skin tissue. These findings highlight the potential of random fibroin fibers/aligned CFs hybrid fibrous scaffold as a promising approach for skin tissue filling and tissue regeneration.

Epimedium sagittatum Maxim ameliorates adriamycin-induced nephropathy by restraining inflammation and apoptosis via the PI3K/AKT signaling pathway.

BACKGROUND: Modern pharmacological studies show that Epimedium sagittatum Maxim (EPI) has antioxidant, antiapoptotic, anti-inflammatory effects. However, the effects of EPI on adriamycin-induced nephropathy are unclear. AIM: The main purpose of this study is to investigate the effects of EPI on adriamycin-induced nephropathy in rats. METHODS: The chemical composition of EPI was detected by high performance liquid chromatography. Network pharmacology was used to collect the effects of EPI on adriamycin nephropathy; renal histological changes, podocyte injury, inflammatory factors, oxidative stress levels, apoptosis levels, and the PI3K/AKT signaling pathway were examined. Moreover, analyze the effects of icariin (the representative component of EPI) on adriamycin-induced apoptosis and PI3K/AKT signaling pathway of NRK-52e cells. RESULTS: Network pharmacological results suggested that EPI may ameliorate adriamycin-induced nephropathy by inhibiting inflammatory response and regulating the PI3K/AKT signaling pathway. The experimental results showed that EPI could improve pathological injury, renal function, podocyte injury, and inhibit inflammation, oxidative stress, apoptosis in adriamycin-induced nephropathy rats through the PI3K/AKT signaling pathway. Furthermore, icariin inhibited adriamycin-induced mitochondrial apoptosis in NRK-52e cells. CONCLUSION: This study suggested that EPI ameliorates adriamycin-induced nephropathy by reducing inflammation and apoptosis through the PI3K/AKT signaling pathway, icariin may be the pharmacodynamic substance basis for this effect.

Porphyromonas gingivalis Outer Membrane Vesicles Promote Apoptosis via msRNA-Regulated DNA Methylation in Periodontitis.

The outer membrane vesicles (OMVs) produced by Porphyromonas gingivalis contain a variety of bioactive molecules that may be involved in the progression of periodontitis. However, the participation of P. gingivalis OMVs in the development of periodontitis has not been elucidated. Here, we isolated P. gingivalis OMVs and confirmed their participation in periodontitis both in vivo and in vitro. Microcomputed tomography (micro-CT) and histological analysis showed that under stimulation with P. gingivalis OMVs, the alveolar bone of rats was significantly resorbed in vivo. We found that P. gingivalis OMVs were taken up by human periodontal ligament cells ([hPDLCs]) in vitro, which subsequently resulted in apoptosis and inflammatory cytokine release, which was accomplished by the microRNA-size small RNA (msRNA) sRNA45033 in the P. gingivalis OMVs. Through bioinformatics analysis and screening of target genes, chromobox 5 (CBX5) was identified as the downstream target of screened-out sRNA45033. Using a dual-luciferase reporter assay, overexpression, and knockdown methods, sRNA45033 was confirmed to target CBX5 to regulate hPDLC apoptosis. In addition, CUT&Tag (cleavage under targets and tagmentation) analysis confirmed the mechanism that CBX5 regulates apoptosis through the methylation of p53 DNA. Collectively, these findings indicate that the role of P. gingivalis OMVs is immunologically relevant and related to bacterial virulence during the development of periodontitis. IMPORTANCE P. gingivalis is a bacterium often associated with periodontitis. This study demonstrates that (i) sRNA45033 in P. gingivalis OMVs targets CBX5, (ii) CBX5 regulates the methylation of p53 DNA and its expression, which is associated with apoptosis, and (iii) a novel mechanism of interaction between hosts and pathogens is mediated by OMVs in the occurrence of periodontitis.

Brassica napus BnaNTT1 modulates ATP homeostasis in plastids to sustain metabolism and growth.

The plastid-localized nucleotide triphosphate transporter (NTT) transports cytosolic adenosine triphosphate (ATP) into plastid to satisfy the needs of biochemistry activities in plastid. Here, we investigate the key functions of two conserved BnaNTT1 genes, BnaC06.NTT1b and BnaA07.NTT1a, in Brassica napus. Binding assays and metabolic analysis indicate that BnaNTT1 binds ATP/adenosine diphosphate (ADP), transports cytosolic ATP into chloroplast, and exchanges ADP into cytoplasm. Thylakoid structures are abnormal and plant growth is retarded in CRISPR mutants of BnaC06.NTT1b and BnaA07.NTT1a. Both BnaC06.NTT1b and BnaA07.NTT1a play important roles in the regulation of ATP/ADP homeostasis in plastid. Manipulation of BnaC06.NTT1b and BnaA07.NTT1a causes significant changes in glycolysis and membrane lipid composition, suggesting that increased ATP in plastid fuels more seed-oil accumulation. Together, this study implicates the vital role of BnaC06.NTT1b and BnaA07.NTT1a in plant metabolism and growth in B. napus.

Activin B-activated Cdc42 signaling plays a key role in regulating adipose-derived mesenchymal stem cells-mediated skin wound healing.

BACKGROUND: In our previous study, activin B in combination with ADSCs enhances skin wound healing. However, the underlying molecular mechanisms are not well studied. Cdc42 is recognized to play a critical role in the regulation of stem cells. METHODS: Pull-down assay was performed to investigate the activity of Cdc42. The dominant-negative mutant of Cdc42 (Cdc42N17) was used to explore the role of Cdc42 in activin B-induced ADSCs migration, proliferation, and secretion in vitro. Cdc42N17-transfected ADSCs were injected into a full-thickness excisional wound model to explore their efficiency in wound healing in vivo. The wound healing efficacy was evaluated by the wound closure rates and histological examination. The neovascularization and wound contraction were detected by immunohistochemistry staining of CD31 and α-SMA. Finally, the underlying mechanisms were explored by RNA sequencing. RESULTS: Cdc42N17 inhibited ADSCs migration, proliferation, and secretion induced by activin B. Furthermore, Cdc42N17-transfected ADSCs inhibited the wound closure rate and suppressed the expression of CD31 and α-SMA induced by activin B in vivo. The RNA sequencing showed that the differentially expressed genes in Cdc42N17-transfected ADSCs versus ADSCs were associated with cell migration, proliferation, and adhesion. Further study revealed that the Cdc42-Erk-Srf pathway was required for activin B-induced proliferation in ADSCs. CONCLUSIONS: Our study indicates that Cdc42 plays a crucial role in ADSCs-mediated skin wound healing induced by activin B.

Adenosine ameliorated Aß25-35-induced brain injury through the inhibition of apoptosis and oxidative stress via an ERα pathway.

Previous studies have shown that adenosine has estrogen-like activity mediated by estrogen receptor α (ERɑ). This study aimed to examine the effects of adenosine on Aß25-35-induced brain injury and the underlying mechanisms involved. Adenosine (Ade, 20 mg/kg, i.g.) was administered for four weeks, followed by the induction of Alzheimer's disease (AD) by Aß25-35 (200 µM, 3 µL/20 g, i.c.v.). Furthermore, a specific ERα blocker (MPP, 0.3 mg/kg, i.p.) was administered 30 min before the treatments of adenosine to evaluate whether the observed effects elicited by adenosine were mediated via ERα. In addition, the learning and memory ability, neuronal damage, and the levels of amyloid ß-protein (Aß), phosphorylated Tau Protein (p-Tau), apoptosis, oxidative stress, immune cells, and ERα were examined. The antagonistic effect of MPP (1 µM) against adenosine (5 µM) in Aß25-35 (20 µM, 24 h)-induced N9 and PC-12 cells was also investigated. Adenosine improved learning and memory ability, reduced neuronal damage, downregulated Aß42/Aß40, p-Tau, apoptosis, and oxidative stress, transformed immune cells, and increased the expression of ERα following Aß25-35 challenge. MPP could block these effects. Moreover, MPP also blocked the effects of adenosine on the levels of apoptosis and reactive oxygen species (ROS) in Aß25-35-induced N9 and PC-12 cells. Adenosine ameliorated Aß25-35-induced brain injury by inhibiting apoptosis and oxidative stress, possibly via an ERα pathway.

BnaNTT2 regulates ATP homeostasis in plastid to sustain lipid metabolism and plant growth in Brassica napus.

The plastid inner envelope membrane-bond nucleotide triphosphate transporter (NTT) transports cytosolic adenosine triphosphate (ATP) into plastid, which is necessary for the biochemical activities in plastid. We identified a chloroplast-localized BnaC08.NTT2 and obtained the overexpressed lines of BnaC08.NTT2 and CRISPR/Cas9 edited double mutant lines of BnaC08.NTT2 and BnaA08.NTT2 in B. napus. Further studies certified that overexpression (OE) of BnaC08.NTT2 could help transport ATP into chloroplast and exchange adenosine diphosphate (ADP) and this process was inhibited in BnaNTT2 mutants. Additional results showed that the thylakoid was abnormal in a8 c8 double mutants, which also had lower photosynthetic efficiency, leading to retarded plant growth. The BnaC08.NTT2 OE plants had higher photosynthetic efficiency and better growth compared to WT. OE of BnaC08.NTT2 could improve carbon flowing into protein and oil synthesis from glycolysis both in leaves and seeds. Lipid profile analysis showed that the contents of main chloroplast membrane lipids, including monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), and phosphatidylglycerol (PG), were significantly reduced in mutants, while there were no differences in OE lines compared to WT. These results suggest that BnaNTT2 is involved in the regulation of ATP/ADP homeostasis in plastid to impact plant growth and seed oil accumulation in B. napus. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-022-01322-8.

Adjunctive effects of laser therapy on somatosensory function and vasomotor regulation of periodontal tissues in patients with periodontitis: A randomized controlled clinical trial.

BACKGROUND: The purpose of this prospective study was to compare the changes in periodontal somatosensory function and microcirculation in patients with periodontitis following initial treatment with scaling and root planing (SRP) with or without adjuvant laser therapy. METHODS: Twenty-four patients suffering from periodontitis were recruited and randomly allocated into a split-mouth design to either SRP combined laser therapy side (test side) or SRP only side (control side). All treatments were performed by the same investigator at a single visit. Laser Doppler Flowmetry (LDF) and Quantitative Sensory Testing (QST) were performed at baseline (W0), 1 week (1W), 2 weeks (2W), and 4 weeks (4W) after treatment on both sides of the attached gingiva of the maxillary lateral incisor. Clinical examination including probing depth (PD) and bleeding on probing (BOP) was performed at W0, 2W, and 4W on both sides. Data were analyzed with two-way analysis of variance. RESULTS: PD and BOP significantly improved after treatment (P <0.001). LDF values were significantly decreased on both sides at all follow-up time points (P <0.001), temperature was increased only on the test side (P = 0.017) whereas there was no significant change on the control side (P = 0.792). Significantly less sensitivity was observed for all QST parameters (P <0.030) except for warmth detection after treatment. CONCLUSION: Adjunctive use of laser therapy did not provide any significant clinical advantage or additional effects on the recovery of periodontal somatosensory function or gingival microcirculation in the present study.