Matrix Metalloproteinase-1 Expression Research Articles

Risk Factors for Pulmonary Metastasis in Differentiated Thyroid Carcinoma Patients and the Significance of Changes in Matrix Metalloproteinase 13 and microRNA-142 Levels.

This work aims to explore the risk factors of lung metastasis (LM) in differentiated thyroid cancer (DTC) (LM-DTC) and the effect of treatment and to detect the relationship between LM-DTC and the levels of matrix metalloproteinase-13 (MMP-13) and micro ribonucleic acid (RNA)-142 (miR-142) in peripheral blood. The data of 420 patients with DTC who are admitted from March 2020 to December 2021 are collected and divided into a non-metastasis group (non-LM group) of 400 cases and metastasis group (LM group) of 20 cases according whether the mung metastasis is found. In addition, risk factors of LM-DTC are analysed and compared. The results of multivariate logistic analysis show that age, disease course, and imaging timing are independent influencing factors of the radionuclide treatment effect. Follicular carcinoma, abnormal expressions of MMP-13, and miR-142 can increase the risk of LM-DTC. MMP-13 and miR-142 can be undertaken as auxiliary diagnostic biological indicators.

Aqueous Extracts of Pleurotus ostreatus and Hericium erinaceus Protect against Ultraviolet A-Induced Damage in Human Dermal Fibroblasts.

Pleurotus ostreatus (PO) and Hericium erinaceus (HE) have been traditionally used to treat various diseases, owing to their antioxidant, antimicrobial, neuroprotective, and antitumor effects. However, few studies have been reported on their antiaging effects. In this study, the antioxidant and antiaging activities of PO and HE aqueous extracts were investigated in ultraviolet A (UVA)-induced human dermal fibroblast cells (HDFs). The antioxidant properties of PO and HE aqueous extracts were measured by total polyphenol and ergothioneine content, and their antioxidant activity was analyzed with the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical-scavenging assays. To demonstrate the antiaging effect of PO and HE aqueous extracts in UVA-induced HDFs, the secretion and mRNA expression of matrix metalloproteinase-1 (MMP-1), procollagen type I (PC1), and elastase were assessed by enzyme-linked immunosorbent assay (ELISA) and real-time PCR, respectively. The total polyphenol content in each extract was 13.6 and 11.7 mg gallic acid equivalents/g dry weight (DW), respectively, and the total ergothioneine content in each extract was 3.43 and 2.18 mg/g DW, respectively. The PO and HE extracts increased DPPH and ABTS radical-scavenging activity in a dose-dependent manner. In UVA-damaged HDFs, the extracts increased PC1 production but decreased MMP-1 production and elastase-1 activity. Furthermore, the mRNA levels of PC1, MMP-1, and elastase were recovered in the PO- and HE-treated UVA-irradiated HDFs compared to those in the irradiated control group. PO and HE aqueous extracts may be potentially used as a promising antiphotoaging agent.

Pectinase halophyte complex extract protects hairless mice skin damaged by UV-irradiation

Abstract The aim of this study was to evaluate the inhibitory effect and mechanism of collagen degradation by Pectinase-assisted Halophyte Complex Extract (PHCE) in hairless mice. Collagen expression in vitro was measured using Enzyme-linked Immunosorbent Assay (ELISA) and Western blotting. After collecting mice skin in vivo, histological analysis was performed with Hematoxylin & Eosin (H & E) and trichrome stain. Molecular expression was measured by western blot analysis. In HaCaT cells, the collagen expression level was increased compared to the positive control group. Epidermal thickness inhibition and collagen expression levels were also increased in the skin of a hairless mouse model. Western blot analysis confirmed the increase in collagen and elastin by inhibiting phosphorylation of NF-κB and decreasing the expression of matrix metalloproteinases-1 (MMP-1) and matrix metalloproteinases-9 (MMP-9). The skin protective effect of PHCE is considered to be due to inhibition of MMPs expression through inhibition of NF-κB activation. Therefore, PHCE can be considered as a candidate material as a food material that can improve skin damage caused by UV rays.

The protective effects of etomidate against interleukin-1β (IL-1β)-induced oxidative stress, extracellular matrix alteration and cellular senescence in chondrocytes

ABSTRACT Osteoarthritis (OA) is a common chronic inflammatory disease associated with aging. Etomidate is an intravenous anesthetic with profound antioxidant and anti-inflammatory effects. We speculated that etomidate might exert a beneficial effect on OA. Herein, we explored the effects of etomidate on interleukin-1β (IL-1β)- induced chondrocytes. Our results prove that etomidate ameliorated the IL-1β-induced oxidative stress in C28/12 chondrocytes by decreasing and increasing the reactive oxygen species (ROS) and glutathione peroxidase (GPx) levels, respectively. Etomidate prevented the IL-1β-induced increase in the expressions of matrix metalloproteinase-3 (MMP-3) and matrix metalloproteinase-13 (MMP-13) in C28/I2 chondrocytes at both mRNA and protein levels. It also caused a significant reduction in the percentage of senescence-associated-β-galactosidase (SA-β-Gal)‐stained chondrocytes, while inducing elevated telomerase activity in IL-1β-treated C28/I2 chondrocytes. The expression levels of senescence regulators, plasminogen activator inhibitor-1 (PAI-1) and p16, were also inhibited by etomidate in IL-1β-treated C28/I2 chondrocytes. In addition, etomidate caused the activation of Adenosine 5ʹ-monophosphate (AMP)-activated protein kinase (AMPK), along with upregulated expression levels of phosphorylated AMPKα and phosphorylated acetyl-Co A carboxylase (ACC). Moreover, blockage of AMPK using compound C abolished the protective effects of etomidate on IL-1β-challenged C28/I2 chondrocytes. Taken together, these results demonstrate that etomidate protected C28/I2 chondrocytes from IL-1β-induced oxidative stress, ECM degradation, and cellular senescence via activating AMPK signaling.

14-3-3 Sigma Protein Contributes to Hepatocyte Growth Factor-mediated Cell Proliferation and Invasion via Matrix Metalloproteinase-1 Regulation in Human Gastric Cancer.

The 14-3-3 protein family has a variety of functions in cellular responses in different organisms, including cell-cycle regulation, apoptosis, and malignant transformation. 14-3-3 Sigma protein (14-3-3σ) induces G2 arrest, which enables repair of damaged DNA. The purpose of this study was to identify the role of 14-3-3σ up-regulation by hepatocyte growth factor (HGF) in cancer cell proliferation and invasion in gastric cancer. In this study, cell culture, western blotting, real-time polymerase chain reaction, zymography, 14-3-3σ knock-down using short hairpin RNA (shRNA), electrophoresis mobility-shift assay, chromatin immunoprecipitation assay and standard two-chamber invasion assay were applied. Firstly, we confirmed that the expression of 14-3-3σ in gastric cancer cells was up-regulated by HGF. To identify how HGF-induced 14-3-3σ expression affects matrix metalloproteinase-1 (MMP1) expression, the cells were treated with the mitogen-activated protein kinase kinase inhibitor PD098059 and analyzed using western blotting. The HGF-mediated expression of MMP1 protein decreased in the presence of PD098059. The role of 14-3-3σ in MMP1 expression was determined through 14-3-3σ knockdown using shRNA. 14-3-3σ-shRNA cells showed reduced levels of MMP1, phosphorylated extracellular signal-regulated kinase, and pp38. HGF-mediated cell proliferation and in vitro invasion were reduced in 14-3-3σ knockdown cells. Serum 14-3-3σ levels were also significantly reduced following gastrectomy in patients with stage II or stage III gastric cancer (p<0.05). These results suggest that 14-3-3σ plays an important role in cell proliferation and metastasis in gastric cancer, and 14-3-3σ may be a novel target for detection and prevention of progression of gastric cancer. In addition, the serum 14-3-3σ level is associated with treatment status in patients with locally advanced gastric cancer.

Effects of temperature-sensitive hydroxybutyl chitosan hydrogel on wound healing of full-thickness skin defect in rats

Effects of temperature-sensitive hydroxybutyl chitosan hydrogel on wound healing of full-thickness skin defect in rats

Topical administration of Tamanu Oil (Calophyllum inophyllum) inhibited the increase of matrix metalloproteinase-1 (MMP-1) expressions and decrease of collagen dermis amount in male wistar rats exposed to ultraviolet B

Background: UV-B irradiation is the major causative factor in photoaging. Photoaging increases MMP-1 significantly and cause damage to collagen by forming ROS. The monounsaturated free fatty acids and antioxidant compounds in Tamanu oil have been studied to inhibit the emergence of MMP-1 and maintain collagen amount. The study aimed to investigate topical Tamanu oil in inhibiting MMP-1 expressions and collagen decrease in male Wistar rats’ skin exposed to UV-B.&#x0D; Methods: A randomized post-test-only control group design was performed on 36 male Wistar rats, aged 2-3 months old with 180-220 grams body weight. They were assigned to two groups: the placebo group and the Tamanu oil group. Both groups were given the same amount of UV-B exposure for four weeks totalling in 840 mJ/cm2 of dosage radiation. Their dermis was examined for MMP-1 using Immunohistochemistry and collagen amount using Picro Sirius Red staining.&#x0D; Results: A comparative analysis was done to show the difference between groups. The mean expression of MMP-1 in the control group was 34,03±2.97 cell/HPF and the Tamanu oil group was lower at 22.73±2.68 cell/HPF. The mean amount of collagen dermis in the control group was 38.90±11.64 % and the Tamanu oil group was higher at 65.66±8.13 %. A significant difference was shown in the two groups (p&lt;0.05).&#x0D; Conclusion: Topical Tamanu oil has proven to hinder the increase of MMP-1 expressions and collagen amount in male Wistar rats’ skin exposed to UV-B.

MicroRNA-98-5p modulates cervical cancer progression via controlling PI3K/AKT pathway

ABSTRACT To probe into the potential mechanism of microRNA (miR)-98-5p inhibiting the biological progress of cervical cancer (CC) cells via regulating PI3K/Akt pathway. Reverse transcription quantitative polymerase chain reaction was applied to detect miR-98-5p expression in CC tissues and cell lines; Cell counting kit-8 and Edu analysis were performed for checking cell proliferation, flow cytometry for cell apoptosis, transwell for cell invasion and migration, Western blot for proliferation-related proteins Ki67 and Proliferating cell nuclear antigen expression, apoptosis-related proteins Bcl-2 and Bax expression, epithelial–mesenchymal transition (EMT)-related proteins Snail, matrix metalloproteinase-3, E-cadherin and N-cadherin expression, as well as PI3K/Akt pathway-related proteins PTEN, PI3K as well as Akt expression levels, and the nude mouse tumor xenograft experiment was applied to verify in vivo. The result clarified, miR-98-5p was reduced in CC. Overexpression miR-98-5p could inhibit CC cell proliferation, invasion, migration and EMT, whereas promoted its apoptosis, but silencing miR-98-5p was opposite. Overexpression miR-98-5p could depress the activation of PI3K/Akt pathway in CC in vivo and in vitro. MiR-98-5p targeted CBX5. In short, miR-98-5p is able to be used as a potential target for treating CC in future research.

Effect of single-dose locally applied lactoferrin on autograft healing in peri-implant bone in rat models

Immediate dental implant installation into fresh extraction sockets has become a common surgical technique and yields successful clinical results. In addition, complete contact may not be possible with this procedure cause of defects between the bone wall and the implant surface. Therefore, different graft materials have been used in the literature to increase the peri‑implant bone volume. The aim of the present study was to evaluate the effect of single-dose and locally applied lactoferrin on autograft healing in peri‑implant area and bone implant contact value. Twenty-four Sprague-Dawley rats were included in this study. Firstly, a trephine drill was used for creating a cylindrical bony defects (6.5 mm in diameter and 3 mm in depth) under sterile saline irrigation in the lateral side of the femur. Subsequently, implant beds –2.5 mm diameter and 6 mm depth – were prepared in the middle of each defect with special implant drills. All of the implants were installed and primary stability was achieved. Rats were randomly divided into 3 groups (n = 8 each): Group-1 had empty defects, Group-2 had defects filled with autograft, and Group-3 had defects filled with autograft and lactoferrin solution (100 μg/ml) combination. All of the rats were sacrificed at postoperative 4th week and samples were analyzed with micro-computed tomography, histomorphometry and immunohistochemistry respectively. It was found that Group 3 had the least area of fibrous tissue (6.75±0.83mm2) according to the other 2 groups (p<0.001). On the other hand, Group 3 had the highest osteoblast number (25.50±3.29), osteoclast number (21.25±1.03), newly formed bone area (20.50±1.30 mm2), total healing area (22.62±0.93 mm2), defect closure rate (80.37±1.40%), bone implant contact value (23.2%±0.6%), and percentage bone volume (18.2%±0.3%) (p<0.001). Matrix metalloproteinase-3 expression was found to be highest in Group 3 by immunohistochemistry analysis. In this study it was observed that the results of the different analysis techniques supported each other. According to these findings it can be stated that a single-dose and locally applied lactoferrin solution plays an important role in the autograft healing in peri‑implant area and increasing bone implant contact value. These findings will shed light on further clinical studies of implant osseointegration.

Effects of Sargassum thunbergii Extract on Skin Whitening and Anti-Wrinkling through Inhibition of TRP-1 and MMPs.

Sargassum thunbergii has been traditionally used as an edible and medicinal material in oriental countries. However, the skin-whitening and anti-wrinkling effects of S. thunbergii have not yet been investigated. This study was conducted to establish optimal extraction conditions for the production of bioactive compounds with antioxidant activity as well as skin-whitening and anti-wrinkle effects using ultrasound-assisted extraction (UAE) in S. thunbergii. The extraction time (5.30~18.7 min), extraction temperature (22.4~79.6 °C), and ethanol concentration (0.0~99.5%), which are the main variables of the UAE, were optimized using a central composite design. Quadratic regression equations were derived based on experimental data and showed a high coefficient of determination (R2 > 0.85), demonstrating suitability for prediction. The optimal UAE condition for maximizing all dependent variables, including radical scavenging activity (RSA), tyrosinase inhibitory activity (TIA), and collagenase inhibitory activity (CIA), was identified as an extraction time of 12.0 min, an extraction temperature of 65.2 °C, and ethanol of 53.5%. Under these conditions, the RSA, TIA, and CIA of S. thunbergii extract were 86.5%, 88.3%, and 91.4%, respectively. We also confirmed S. thunbergii extract had inhibitory effects on the mRNA expression of tyrosinase-related protein-1, matrix metalloproteinase-1, and matrix metalloproteinase-9, which are the main genes of melanin synthesis and collagen hydrolysis. Liquid chromatography-tandem mass spectrometry was used to identify the main phenolic compounds in S. thunbergii extract, and caffeic acid was identified as a major peak, demonstrating that high value-added ingredients with skin-whitening and anti-wrinkling effects can be produced from S. thunbergii and used for developing cosmetic materials.

Orthodontic force regulates metalloproteinase-3 promoter in osteoblasts and transgenic mouse models

Background/purposePreviously we demonstrated up-regulation of matrix metalloproteinase-3 (MMP-3) in human osteoblasts under compression and in bony specimens of experimental orthodontic tooth movement (OTM). Here, we studied the temporal characteristics of compression stimulation in human and mouse osteoblast cell lines, and generated a transgenic mouse model for assessing the MMP-3 expression during OTM. Materials and methodsWe investigated MMP-3 expressions in human and murine osteoblasts through RT-PCR and luciferase assay, after compressive force loading. Inhibitors were added to identify the possible mechanisms for signal transduction. A human MMP-3 promoter was isolated, cloned and transfected to generate a transgenic mouse with a green fluorescent protein reporter. OTM was then initiated to observe the location and time course of transcriptional regulation of MMP-3 signals. ResultsWe found changes in the transcription of MMP-3 in response to mechanical force applied to both human and mouse osteoblast cell lines, suggesting that the response is positive across species. Cloned human MMP-3 promoter may cause the response of luciferase to 1% compression. Moreover, p38 inhibitor exerted a down-regulatory effect on MMP-3 promoter expression, although the inhibitory effect didn't reach a significant level. In the transgenic mouse OTM model, we again found increased expression of MMP-3 in response to mechanical force loading around the periodontal ligament. ConclusionMechanical force can stimulate MMP-3 expression, possibly through the p38 MAPK pathway, with its strongest signal occurring at 24 h. The mechanical responsiveness in MMP-3 promoter regions can be observed in both humans and rodents in vitro and in vivo.

Warming moxibustion attenuates inflammation and cartilage degradation in experimental rabbit knee osteoarthriti.

To explore the molecular mechanism of warming moxibustion (WM) in knee osteoarthritis (KOA). The knee joints of 40 New Zealand rabbits were placed in a plaster cast in an extended position to establish a KOA model. The animals were randomly divided into four groups: the control group, model group, WM group, and diclofenac (DF) group. Hematoxylin-eosin staining and the modified Mankin score were applied to evaluate the histopathological changes. Chondrocyte apoptosis was determined by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Western blotting and real-time quantitative polymerase chain reaction were performed to measure the expression of interleukin-1α (IL-1β), prostaglandin E receptor 3 (PTGER3), a disintegrin-like and metalloproteinase with thrombospondin type-1 motifs-5 (ADAMTS-5), matrix metalloproteinase-13 (MMP-13), and C-terminal telopeptides of collagen type II (CTX-II) in cartilage tissues of the different groups. The concentrations of IL-1β, PTGER3, and CTX-II in serum were detected by the enzyme-linked immunosorbent assay. Rabbits with KOA in the WM and DF groups showed significantly reduced cartilage erosion and Mankin scores, compared with the untreated rabbits. The number of TUNEL-positive cells observed in the WM group was much fewer than that in the model group. The expression of PTGER3, MMP-13, CTX-II IL-1β, and ADAMTS-5 in cartilage tissues was remarkably downregulated following therapy with WM and DF. Moreover, a marked reduction was observed in the serum levels of IL-1β, PTGER3, and CTX-II in the WM and DF groups. WM exerts favorable therapeutic effects on articular injuries of KOA by regulating the expression of inflammatory and cartilage degradation-related cytokines.

MicroRNA-4423-3p inhibits proliferation of fibroblast-like synoviocytes by targeting matrix metalloproteinase 13 in rheumatoid arthritis

ABSTRACT Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease that is increasing in incidence worldwide. RA is regulated by a variety of microRNAs (miRNAs/miR). Moreover, analysis of public data has revealed that miR-4423-3p is significantly downregulated in peripheral blood mononuclear cells of RA patients. This study investigated the role of miR-4423-3p in RA. The levels of miR-4423-3p and matrix metalloproteinase 13 (MMP13) in RA patients and the regulatory relationship between miR-4423-3p and MMP13 were analyzed using public data. A dual-luciferase reporter assay was performed to verify that miR-4423-3p targets MMP13 in human fibroblast-like synoviocyte (HFLS) RA cells (HFLS-RA). Following the overexpression of miR-4423-3p, miR-4423-3p inhibitor, and MMP13 in HFLS-RA, viability, proliferation, cell cycle, apoptosis, and invasion/migration assays were used to detect the effects of miR-4423-3p targeting MMP13 on cell biological processes. The results revealed that miR-4423-3p was downregulated in peripheral blood mononuclear cells of RA patients and MMP13 was upregulated in synovial tissue of RA patients. miR-4423-3p targets the 3ʹ untranslated region of MMP13 and downregulates MMP13 expression. After overexpression of miR-4423-3p, cell proliferation, migration, and invasion were inhibited, the cell cycle was prevented and cell apoptosis was promoted. Overexpression of MMP13 promoted cell proliferation, migration, and invasion, while accelerating the cell cycle process and suppressing apoptosis. The findings indicate that in HFLS-RA cells, overexpression of miR-4423-3p inhibited proliferation, migration, and invasion, and promoted apoptosis by negatively regulating MMP13. The overexpression of miR-4423-3p might be a novel strategy for the treatment of RA.

CircPan3 Promotes the Ghrelin System and Chondrocyte Autophagy by Sponging miR-667-5p During Rat Osteoarthritis Pathogenesis

This study aimed to investigate the potential roles of circRNAs in regulating osteoarthritis (OA)-related ghrelin synthesis, autophagy induction, and the relevant molecular mechanisms. Results showed that Col2a1, Acan, ghrelin, and autophagy-related markers expression were downregulated, while matrix metalloproteinase 13 (MMP13) and a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5) expressions increased in both IL-1β-induced rat chondrocytes and cartilage tissues of OA rats. A total of 130 circRNAs and 731 mRNAs were differentially expressed in IL-1β-induced rat chondrocytes. Among them, we found that circPan3 expression was significantly decreased in both cellular and animal OA models. CircPan3 directly targeted miR-667-5p. CircPan3 overexpression promoted Col2a1, Acan, ghrelin, beclin 1, and LC3-II expression but reduced MMP13 and ADAMTS5 expression in rat chondrocytes, whereas overexpression of miR-667-5p exhibited opposite effects on the above markers. Furthermore, we found that miR-667-5p bound directly to the 3′-UTR sequence of ghrelin gene. Moreover, the circPan3-induced alterations in chondrocytes were antagonized by miR-667-5p overexpression. Taken together, our findings demonstrate that circPan3 promotes ghrelin synthesis and chondrocyte autophagy via targeting miR-667-5p, protecting against OA injury. This study provided experimental evidence that circPan3/miR-667-5p/ghrelin axis might serve as targets of drug development for the treatment of OA.

Circular RNA RBM33 contributes to extracellular matrix degradation via miR-4268/EPHB2 axis in abdominal aortic aneurysm.

BackgroundAbdominal aortic aneurysm (AAA) is a complex vascular disease involving expansion of the abdominal aorta. Extracellular matrix (ECM) degradation is crucial to AAA pathogenesis, however, the specific molecular mechanism remains unclear. This study aimed to investigate differentially expressed circular RNAs (DEcircRNAs) involved in ECM degradation of AAA.MethodsTranscriptome sequencing was used to analyze the DEcircRNAs between the AAA tissues and normal tissues. The expression of circRNAs in tissues and cells was validated using quantitative reverse transcription PCR (RT-qPCR). Overexpression of circRNAs in vascular smooth muscle cells (VSMCs) treated with angiotensin II (Ang II) was employed to explore its effect on ECM degradation of AAA. Bioinformatic technology, luciferase reporter gene assay, RT-qPCR, and rescue experiment were employed to evaluate the regulatory mechanism of circRNA.ResultsWe identified 65 DEcircRNAs in AAA tissues compared with normal abdominal aortic tissues, including 30 up-regulated and 35 down-regulated circRNAs, which were mainly involved in inflammation and ECM-related functions and pathways. Moreover, circRBM33 was significantly increased in AAA tissues and Ang II-induced VSMCs compared with control samples. Overexpression of circRBM33 increased the expression of ECM-related molecule matrix metalloproteinase-2 and reduced the tissue inhibitor of matrix metalloproteinases-1 expression. Mechanistically, miR-4268 targeted binding to circRBM33 and inhibited the luciferase activity of circRBM33. Overexpression of circRBM33 induced the expression of EPH receptor B2 (EPHB2), and this effect was countered by miR-4268 mimics.ConclusionsOverall, our data suggest that circRBM33 might be involved in AAA progression by regulating ECM degradation via the miR-4268/EPHB2 axis.

Oxidative stress as a critical factor might involve in intervertebral disc degeneration via regulating NOXs/FOXOs

BackgroundOxidative stress is involved in many musculoskeletal diseases, such as osteoarthritis. However, the effect of oxidative stress on intervertebral disc degeneration (IDD) is still unclear. This study was aimed to provide an evidence of oxidative stress involved in IDD, and propose a new insight into pathogenesis of IDD. MethodsSixteen rats were randomly divided into sham and cervical muscle section (CMS) groups. The intervertebral disc degeneration scores (DDS) were assessed by histological staining at 8 weeks. Intracellular reactive oxygen species mainly comes from nicotinamide adenine dinucleotide phosphate oxidases (NOXs), while its clearance relies on antioxidant enzymes which regulated by forkhead transcription factor O (FOXOs). Thus, the oxidative stress was evaluated by the expression of NOXs and FOXOs. Meanwhile, the protein expression of Aggrecan, matrix metalloproteinase-13 (MMP-13), NOXs, FOXOs and antioxidant proteins (Manganese superoxide dismutase: MnSOD and Catalase) were tested in nucleus pulposus cells (NPCs) under tert-butyl hydroperoxide (TBHP) intervention. ResultsCMS induced IDD by enhancing DDS in 8 weeks, and the expression of NOX2 and NOX4 were significantly increased and the expression of FOXO3 and FOXO4 were remarkably decreased in the CMS rats. With the stimulation of TBHP, the contents of NOX2 and NOX4 in NPCs increased significantly, and the antioxidant proteins of FOXO1, FOXO3, FOXO4, MnSOD and Catalase and the matrix proteins of Aggrecan decreased remarkably, while MMP-13 significantly increased after TBHP intervention. ConclusionsThe present study proposed that regulation of NOXs and FOXOs alters oxidative stress in intervertebral disc, which indicates that the intervention of oxidative stress would provide a new strategy to the treatment of IDD.

Aspirin Exerts Neuroprotective Effects by Reversing Lipopolysaccharide-Induced Secondary Brain Injury and Inhibiting Matrix Metalloproteinase-3 Gene Expression.

Objective This study is aimed at exploring the possible neuroprotective mechanism of aspirin and the effect of bacterial endotoxin lipopolysaccharide (LPS) during cerebral ischaemia-reperfusion (CIRP) injury. Methods We established three animal models: the CIRP, LPS, and CIRP+LPS models. Mortality, the injured brain area, and the beam walking test were used to estimate the degree of cerebral injury among the rats. Immunohistochemistry and immunofluorescence were used to detect activated microglia, matrix metalloproteinase-3 (MMP-3), and osteopontin (OPN). Results The injured brain area and mortality were dramatically reduced (p < 0.01), and the beam walking test scores were elevated (p < 0.01) in the acetylsalicylic acid (ASA) group compared to the control group. The number of microglia-, MMP-3-, and OPN-positive cells also increased. Furthermore, the number of GSI-B4, OPN, and MMP-3 cells decreased in the ASA group compared to the control group. After LPS stimulation, the number of microglia reached a peak at 24 h; at 7 d, these cells disappeared. In the ASA group, the number of microglia was significantly smaller (p < 0.05), especially at 24 h (p < 0.01), compared to the LPS group. Moreover, the injured brain area and the mortality were dramatically increased and the beam walking test scores were reduced (p < 0.01) after LPS simulation following CIRP. The degree of injury in the ASA group resembled that in the control group. However, the number of MMP-3-immunoreactive neurons or microglia was significantly larger than that of the control group (p < 0.05). In the ASA group, the MMP-3 expression was also considerably decreased (p < 0.05). Conclusions After CIRP, microglia were rapidly activated and the expression of MMP-3 and OPN significantly increased. For rats injected with LPS at reperfusion, the injured brain area and mortality also dramatically increased and the neurologic impairment worsened. However, ASA exhibited a neuroprotective effect during CIRP injury. Furthermore, ASA can reverse LPS-induced cerebral injury and inhibit the inflammatory reaction after CIRP injury.

Sesquiterpenes From Oplopanax elatus Stems and Their Anti-Photoaging Effects by Down-Regulating Matrix Metalloproteinase-1 Expression via Anti-Inflammation.

In the process of continuing to investigate ultraviolet b (UVB) irradiation protective constituents from Oplopanax elatus stems, nine new sesquiterpenes, named as eurylosesquiterpenosides A–D (1–4), eurylosesquiterpenols E–I (5–9), and ten known ones (10–19) were gained. Their structures were established by analysis of their NMR spectroscopic data, and electronic circular dichroism calculations were applied to define their absolute configurations. In addition, UVB induced HaCaT cells were used to study their anti-photoaging activities and mechanism. The results consolidated that compounds 7, 11, and 14 could improve the survival rate of HaCaT cells in concentration dependent manner at 10, 25, and 50 μM. Furthermore, western blot assay suggested that all of them could inhibit the expression of matrix metalloproteinase-1 (MMP-1), and increase the level of type I collagen markedly. Compounds 11 and 14 could reduce the phosphorylation of extracellular signal-regulated kinase and p38, respectively. Besides, compounds 7, 11, and 14 could significantly down-regulate the expression of inflammation related protein, such as tumor necrosis factor-α and cyclooxygenase-2, which indicated that they played anti-photoaging activities by reducing MMP-1 expression via down-regulating the production of inflammatory mediators and cytokines in UVB-induced HaCaT cells.

English

Abeliophyllum distichum Nakai is a deciduous shrub that belongs to Oleaceae Abeliophyllum and grows only in Korea. In this study, the antioxidative activity, cytotoxicity, anti-aging activity, and levels of acteoside and isoacteoside were compared between non-fermented and fermented (using Lactobacillus plantarum and Lactobacillus brevis) extracts of A. distichum Nakai. The cytotoxicity of these extracts, along with their ability to inhibit elastase activity and Matrix metalloproteinase-1 (MMP-1) expression, and their ability to promote type I procollagen synthesis were investigated in human dermal fibroblasts cells. These tests revealed that the fermented extract possessed higher antioxidant and anti-aging activities compared with the non-fermented extract. The levels of acteoside and isoacteoside were about 1.25 and 1.05 times higher in the fermented extract than in the non-fermented extract. It was speculated that they were converted from acteoside glucosides and isoacteoside glucosides via bioconversion by the fermentation strains. Together, these findings indicate that extracts of A. distichum Nakai show good potential as antioxidative and anti-aging cosmetic materials. &nbsp; Key words: Abeliophyllum distichum Nakai, Lactobacillus plantarum, Lactobacillus brevis, fermentation, antioxidative activity, anti-aging activity.

Amelioration of Posttraumatic Osteoarthritis in Mice Using Intraarticular Silencing of Periostin via Nanoparticle-Based Small Interfering RNA.

Recent evidence delineates an emerging role of periostin in osteoarthritis (OA), since its expression after knee injury is detrimental to the articular cartilage. We undertook this study to examine whether intraarticular (IA) knockdown of periostin would ameliorate posttraumatic OA in a murine model. Posttraumatic OA was induced in 10-week-old male C57BL/6J mice (n = 24) by destabilization of the medial meniscus (DMM), and mice were analyzed 8 weeks after surgery. Periostin expression was inhibited by small interfering RNA (siRNA) delivered IA using a novel peptide-nucleotide polyplex. Following histologic assessment of the mouse knee cartilage, the extent of cartilage degeneration was determined using Osteoarthritis Research Society International (OARSI) cartilage damage score, and severity of synovitis was also assessed. Bone changes were measured using micro-computed tomography. The effect and mechanism of periostin silencing were investigated in human chondrocytes that had been stimulated with interleukin-1β (IL-1β) with or without the IκB kinase 2 inhibitor SC-514. Periostin expression in mice with posttraumatic OA was significantly abolished using IA delivery of a peptide-siRNA nanoplatform. OARSI cartilage damage scores were significantly lower in mice receiving periostin siRNA (mean ± SEM 10.94 ± 0.66) compared to untreated mice (22.38 ± 1.30) and mice treated with scrambled siRNA (22.69 ± 0.87) (each P = 0.002). No differences in the severity of synovitis were observed. Subchondral bone sclerosis, bone volume/total volume, volumetric bone mineral density, and heterotopic ossification were significantly lower in mice that had received periostin siRNA treatment. Immunostaining of cartilage revealed that periostin knockdown reduced the intensity of DMM-induced matrix metalloproteinase 13 (MMP-13) expression and also diminished the phosphorylation of p65 and immunoreactivity of the aggrecan neoepitope DIPEN. Periostin knockdown also suppressed IL-1β-induced MMP-13 and ADAMTS-4 expression in chondrocytes. Mechanistically, periostin-induced MMP-13 expression was abrogated by SC-514, demonstrating a link between periostin and NF-κB. IA delivery of the periostin-siRNA nanocomplex represents a promising clinical approach to mitigate the severity of joint degeneration in OA. Our findings may thus provide an unequivocal scientific rationale for longitudinal studies of this approach. Utilizing a cartilage-specific gene-knockout strategy will further illuminate the functional role of periostin in OA.