MMP-1 Expression Research Articles

Myosin heavy chain 2 (MYH2) expression in hypertrophic chondrocytes of soft callus provokes endochondral bone formation in fracture

AimsMuscle–bone interactions during fracture healing are rarely known. Here we investigated the presence and significance of myosin heavy chain 2 (MYH2), a component of myosin derived from muscles, in fracture healing. Main methodsWe collected five hematoma and seven soft callus tissues from patients with distal radius fractures patients, randomly selected three of them, and performed a liquid chromatography–mass spectrometry (LC-MS) proteomics analysis. Proteomic results were validated by histological observation, immunohistochemistry, and immunofluorescence for MYH2 expression. These findings were further confirmed in a murine femoral fracture model in vivo and investigated using various methods in vitro. Key findingsThe LC-MS proteomics analysis showed that MYH proteins were enriched in human soft calluses compared to hematoma. Notably, MYH2 protein is upregulated as high rank in each soft callus. The histological examination showed that MYH2 expression was elevated in hypertrophic chondrocytes within the human soft callus. Consistent with human data, Myh2 were significantly co-localized with Sox9 in hypertrophic chondrocytes of murine femoral fracture, in comparison to pre-hypertrophic and proliferating chondrocytes. Soluble MYH2 protein treatment increased MMP13 and RUNX2 expression in chondrocytes. In soluble MYH2 treatment, proliferation of chondrocytes was not altered, but the osteogenic and chondrogenic features of chondrocytes increased and decreased during differentiation, respectively. SignificanceThese findings indicate the potential of soluble MYH2 protein as a promising therapeutic strategy for promoting endochondral bone formation in chondrocytes following fracture.

Emodin alleviates joint inflammation and bone erosion in rats with collagen-induced arthritis by inhibiting ferroptosis and degrading matrix metalloproteinases

To investigate the osteoprotective mechanism of emodin in light of the ferroptosis signaling pathway in a rat model of rheumatoid arthritis. SD rat models of collagen-induced arthritis (CIA) were treated with methotrexate or low or high doses of emodin, and the changes in arthritis scores and toe volume were recorded. model of CIA rats. Malondialdehyde (MDA) content in the joint cartilage was determined, and ankle joint tissue pathologies were observed using caffein solid green staining and hematoxylin-spermine red staining. MMP3 and MMP13 expressions in the ankle joint tissues were detected using immunohistochemistry, and Western blotting was used to detect the protein expressions of ACSL4, SLC7A11, GPX4, and FTH1. Compared with the normal control rats, the CIA rats showed significantly increased arthritis score index with obvious toe swelling (P<0.05), rough cartilage surface, and loss and irregular aligment of chondrocytes. The rat models also showed significantly increased MDA and ACSL4 contents, lowered SLC7A11, GPX4, and FTH1 contents (P<0.05), and decreased expressions of MMP3 and MMP13 in the ankle joint (P<0.05). The rat models treated with either methotrexate or emodin (40 and 80 mg/kg) had significantly reduced arthritis score index and toe swelling with smooth cartilage surface and neat arrangement of the chondrocytes. The treatments with methotrexate and emodin both decreased the contents of MDA and ACSL4 and increased the contents of SLC7A11, GPX4, and FTH1 in the joint tissues (P<0.05). Emodin can effectively control joint inflammation and improve joint bone erosion in CIA rats possibly by inhibiting the ferroptosis signaling pathways and reducing the expressions of MMP3 and MMP13.

The Effects of Injectable Platelet-Rich Fibrin and Advanced-Platelet Rich Fibrin on Gingival Fibroblast Cell Vitality, Proliferation, Differentiation.

Injectable Platelet Rich Fibrin (I-PRF) and Advanced-Platelet Rich Fibrin (A-PRF) are autologous materials derived from patients' blood and employed in periodontal regenerative surgery. Although I-PRF and A-PRF have different characteristics, their biological effects on gingival tissue fibroblasts remain unclear. This research aims to compare the in vitro capacity in inducing gene expression and proliferation of human gingival fibroblasts between A-PRF and I-PRF. Human donors undergoing dental implant surgery were sampled for normal human gingival fibroblasts (NHGFCs), followed by preparing A-PRF and I-PRF membranes. Enzyme-linked immunosorbent assay (ELISA) kit was used to assess the release of platelet-derived growth factor-AA (PDGF-AA), transforming growth factor-beta1 (TGF- β1), and insulin growth factor-1 (IGF-1) at different periods. Cell viability and proliferation of A-PRF and I-PRF were compared using CCK-8 assay. The impacts of platelet concentration on human gingival fibroblast cells (HGFCs) were evaluated by quantifying the level or amount of phosphorylated extracellular signal-regulated protein kinase (p-ERK), and Matrix metalloproteinases (MMPs), MMP-1 and MMP-3. The effects of PRF on aged human gingival fibroblast cells were examined retrospectively. Overall, A-PRF demonstrated a higher release of TGF-B1 and PDGF-AA, while I-PRF reflected higher levels of IGF-1. A significantly higher level of cell proliferation was induced by higher cell proliferation by A-PRF and I-PRF. Additionally, in comparison to I-PRF, the expression of ERK phosphorylation and MMP-1 &MMP-3 in HGFCs was demonstrated by I-PRF and A-PRF. The increase in A-PRF was time-dependent (p < 0.05). Both I-PRF and A-PRF induced a stimulatory biological impact on the proliferation of human gingiva fibroblasts, with the latter demonstrating better capacity in facilitating the release of different growth factors. A-PRF also induced higher gene expression of p-ERK, MMP-1 &MMP-3, and the proliferation of fibroblasts.

Ubiquitin-specific peptidase 25 exacerbated osteoarthritis progression through facilitating TXNIP ubiquitination and NLRP3 inflammasome activation

Several members of the ubiquitin-specific proteases (USPs) family have been revealed to regulate the progression of osteoarthritis (OA). The current study aimed to investigate the role and the underlying mechanism of USP25 in IL-1β-induced chondrocytes and OA rat model. It was discovered that IL-1β stimulation upregulated USP25, increased ROS level, and suppressed cell viability in rat chondrocytes. Besides, USP25 knockdown alleviated IL-1β-induced injury by decreasing ROS level, attenuating pyroptosis, and downregulating the expression of IL-18, NLRP3, GSDMD-N, active caspase-1, MMP-3, and MMP-13. Furthermore, we discovered that USP25 affected the IL-1β-induced injury in chondrocytes in a ROS-dependent manner. Moreover, USP25 was revealed to interact with TXNIP, and USP25 knockdown increased the ubiquitination of TXNIP. The pro-OA effect of USP25 abundance could be overturned by TXNIP suppression in IL-1β-induced chondrocytes. Finally, in vivo experiment results showed that USP25 inhibition alleviated cartilage destruction in OA rats. In conclusion, we demonstrated that USP25 stimulated the overproduction of ROS to activate the NLRP3 inflammasome via regulating TXNIP, resulting in increased pyroptosis and inflammation in OA.

The expression of matrix metalloproteinase 2, 9 and 11 in Ethiopian breast cancer patients

IntroductionMatrix metalloproteinases (MMPs) play a pathophysiological role in cancer initiation and progression. Numerous studies have examined an association between MMP-2, MMP-9, and MMP-11 expression and clinicopathological characteristics of breast cancer (BC); however, no research has been done on the MMP expression levels in BC cases from Ethiopia.Materials and methodsA total of 58 formalin-fixed paraffin-embedded breast tissue samples encompassing 16 benign breast tumors and 42 BC were collected. The RNA was extracted and quantitative reverse-transcription PCR was performed. GraphPad Prism version 8.0.0 was used for statistical analysis.ResultsThe MMP-11 expression levels were significantly higher in breast cancer cases than in benign breast tumors (P = 0.012). Additionally, BC cases with positive lymph nodes and ER-positive receptors had higher MMP-11, MMP-9, and MMP-2 expression than cases with negative lymph nodes and ER-negative, respectively. The MMP-11 and MMP-9 expressions were higher in grade III and luminal A-like tumors than in grade I-II and other subtypes, respectively.ConclusionThe MMP-11 expression was higher in BC than in benign breast tumors. Additionally, MMP-11, MMP-9, and MMP-2 were higher in BC with positive lymph nodes and estrogen receptors. Our findings suggest an important impact of MMPs in BC pathophysiology, particularly MMP-11.

Kruppel-Like Factor 5 Acetylation Upregulates Runx2 Expression and Accelerates the Dedifferentiation Process of Chondrocyte in Monolayer Culture

This study aimed to investigate the role of KLF5 (Kruppel-like factor 5) in the abnormal dedifferentiation of chondrocytes (CHs) observed in osteoarthritis (OA) and monolayer culture conditions. Human primary CHs were cultured in a monolayer for 14 days, and various analyses were conducted. The area and aspect ratio of the cells were measured, and protein expression levels of KLF5, acetylated KLF5 (Ac-KLF5), and Runx2 were assessed using western blotting. Additionally, mRNA levels of chondrogenic genes (type II collagen, Col-II), hypertrophic genes (type X collagen, Col-X), and MMP-13 were determined through RT-PCR. The effects of TGF-β and Am80 supplementation on KLF5 acetylation and Runx2 expression were examined, and siRNA was used to silence Runx2 gene expression. Results showed that CHs exhibited dedifferentiation after 7 days of culture, characterized by increased cell size, larger aspect ratio, elevated Ac-KLF5 levels, decreased Col-II expression, and increased Col-X and MMP-13 expression. TGF-β treatment enhanced Ac-KLF5 and Runx2 expression, thereby accelerating dedifferentiation within 3 days. In contrast, Am80 suppressed Ac-KLF5 and Runx2 expression, leading to delayed dedifferentiation over 14 days. Silencing Runx2 mitigated KLF5 acetylation-induced CH dedifferentiation without affecting Ac-KLF5 levels. In conclusion, KLF5 acetylation promotes Runx2 expression and subsequently contributes to CH dedifferentiation during monolayer culture.

Correlation Between Synovial Fluid Levels of Matrix Metalloproteinase's (MMP-1, MMP-3, and MMP-9) and TNF-α with the Severity of Osteoarthritis Knee in Rural Indian Population.

In India, 22% to 39% of the ageing population suffers from degenerative knee osteoarthritis (OA), making it the most prevalent joint disorder in the knee. MMP-1, MMP-3, and MMP-9 protein expression levels have all been associated with OA. The aim of the present study was to establish a relationship between synovial fluid levels of matrix metalloproteinases (MMP-1, MMP-3, and MMP-9), and tumour necrosis factor-alpha (TNF-α) with different Kellgren-Lawrence Grading scale as per the severity of knee osteoarthritis (OA). This hospital-based observational study included 87 individuals with knee osteoarthritis examined at the orthopaedics department outpatient clinic at the tertiary care teaching hospital in rural area of north India. In-person interviews were conducted to gather data, through a semi-structured, pretested interview schedule. To determine the degree and severity of OA, the levels of the enzymes matrix metalloproteinase MMP-1, MMP-3, and MMP-9 as well as tumour necrosis factor (TNF-α) were assessed in the synovial fluid of knee of each study participant. The levels of MMP-1, MMP-3, MMP-9, and TNF-α in synovial fluid were significantly correlated with the severity of osteoarthritis as determined by the Kellgren-Lawrence Grading Scale. Age, duration of symptoms and BMI showed a strong positive and significant correlation with biochemical markers (MMP-1, MMP-3, MMP-9, and TNF-α) in synovial fluid of Knee. Level of biochemical markers (MMP-1, -3, -9, and TNF-α) in synovial fluid act as diagnostic markers and have a positive correlation with the severity of osteoarthritis knee, age, weight/BMI and duration of disease. However, no significant correlation was found between the level of aforementioned biochemical markers with sex, height, inflammation of the knee, morning stiffness, and age of onset of disease.

Calcipotriol suppresses GPX4-mediated ferroptosis in OA chondrocytes by blocking the TGF-β1 pathway

Globally, tens of millions of individuals experience osteoarthritis (OA), a degenerative joint condition for which a definitive cure is currently lacking. This condition is characterized by joint inflammation and the progressive deterioration of articular cartilage. In this study, western blotting, quantitative reverse-transcription polymerase chain reaction, and immunofluorescence analysis were performed to elucidate the molecular mechanisms by which calcipotriol alleviates chondrocyte ferroptosis. The effect of calcipotriol on reactive oxygen species and lipid peroxidation levels in chondrocytes was assessed using dihydroethidium staining and the fluorescent dye BODIPY. To replicate OA, the destabilized medial meniscus model was employed, followed by the injection of calcipotriol into the knee articular cavity. Morphological analysis was conducted through hematoxylin and eosin staining, safranin O-Fast green staining, and micro-computed tomography analysis. Immunohistochemical analysis was performed to validate the effect of calcipotriol in vivo. Our results demonstrate that the expression of SOX9, col2a1, and Aggrecan, as well as MMP13 and ADAMTS5 protein expression levels, decrease upon treatment with calcipotriol in interleukin-1β stimulated chondrocytes. Despite these promising outcomes, the exact mechanism underlying calcipotriol's therapeutic effect on OA remains uncertain. We discovered that calcipotriol inhibits chondrocyte GPX4-mediated ferroptosis by suppressing the expression of transforming growth factor-β1. Furthermore, our study established an in vivo model of OA using rats with medial meniscus instability. Our experiments on rats with OA revealed that intra-articular calcipotriol injection significantly reduces cartilage degradation caused by the disease. Our findings suggest that calcipotriol can mitigate OA by impeding GPX4-mediated ferroptosis of chondrocytes, achieved through the suppression of the TGF-β1 pathway.

GKT137831 in combination with adipose-derived stem cells alleviates high glucose-induced inflammaging and improves diabetic wound healing.

Adipose-derived stem cells (ADSCs) have been proven to promote healing in diabetic wounds, which are one of the most serious chronic refractory wounds. However, reactive oxygen species (ROS) induced by high glucose (HG) lead to oxidative stress and aging in ADSCs, which limits the therapeutic effect of ADSCs. In this study, we investigated the role of GKT137831, a NOX1/4 inhibitor that can reduce ROS production, in protecting ADSCs from hyperglycemia and in diabetic wound healing. In vitro, ROS levels and NOX4 expression were increased after HG treatment of ADSCs, while the oxidative stress marker malondialdehyde was increased; mitochondrial membrane potential was decreased; inflammatory aging-related indicators such as p16, p21, matrix metalloproteinase-1 (MMP1), MMP3, interleukin-6, and β-galactosidase were increased; and migration was weakened. In vivo, we constructed a diabetic mouse wound model and found that the combination of ADSCs and GKT137831 synergistically promoted the 21-day wound healing rate, increased the expression of collagen and hydroxyproline, increased the number of blood vessels and the expression of CD31, and reduced the expression of interleukin-6, MMP1, MMP3, and p21. These results suggest that GKT137831 could protect ADSCs from oxidative stress and aging induced by HG and enhance the therapeutic effect of ADSCs on diabetic wounds.

Achyranthes bidentate extracts protect the IL-1β-induced osteoarthritis of SW1353 chondrocytes

Osteoarthritis, the most common joint disease worldwide, is a degenerative disease characterized by cartilage degeneration and inflammation. The active ingredients in the traditional Chinese medicinal plant Achyranthes bidentate can be used to treat waist, leg, and joint pain caused by rheumatism arthralgia. In this study, we identified the optimal microwave extraction protocol for saponins from A.bidentate, evaluated their protective effects against IL-1β-induced inflammation in SW1353 human chondrocytes, and explored their protective pathway. The microwave-extraction parameters required to obtain the maximum yield of A.bidentate saponins using 80% ethanol were identified using response surface methodology. The parameters were solid-liquid ratio, 1:10; extraction time, 20min; power, 721W; temperature, 65°C. The actual yield of saponins extracted was to be 194.01μg/mg extract. The SW1353cells were pretreated with A.bidentate extract (ABE) at a concentration of 50 or 100μg/mL for 3h, after which an inflammatory response was stimulated using IL-1β. The ABE significantly reduced the expression of proinflammatory factors IL-6, TNF-α, COX-2, iNOS, PGE2, and NO, and inhibited NF-κB activity, effectively attenuating the inflammatory response. ABE also inhibited MMP13 and ADAMTS-5 expression, reducing IL-1β-induced degradation of the extrachondral matrix. This confirmed that ABE effectively inhibits NF-κB activity and reduces IL-1β-induced inflammation, extracellular matrix degradation, and expression of apoptotic proteins Bax and caspase-3. Therefore, ABE has potential as a new botanical drug for preventing osteoarthritis.

Anti-Inflammatory Effects of Boric Acid in Treating Knee Osteoarthritis: Biochemical and Histopathological Evaluation in Rat Model.

This study aimed to examine the anti-inflammatory properties of boric acid (BA) in treating knee osteoarthritis (KOA) in rats, evaluating its biochemical and histopathological therapeutic effects.A KOA rat model was induced by injecting monosodium iodoacetate into the knee joint. Random assignment was performed for the experimental groups as follows: group-1(control), group-2(KOA control), group-3 (BA:4mg/kg, orally), group-4(BA:10mg/kg, orally), group-5(BA:4mg/kg, intra-articularly), and group-6(BA:10mg/kg, intra-articularly). The rats received 100 µL of BA intra-articularly on days 1, 7, 14, and 21 or 1mL orally once a day (5days/week) for 4weeks. Serum levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and activityofmatrix metalloproteinase-13 (MMP-13) were measured. Histopathological and immunohistochemical analyses were performed on knee joint samples using specific antibodies for IL-1β, TNF-α, MMP-13, and nitric oxide synthase-2 (NOS-2).Group-2 exhibited higher serum IL-1β and TNF-α levels and MMP-13 activity than group-1 (P < 0.05). However, IL-1β and TNF-α levels and MMP-13 activity were lower in all treatment groups than in group-2, with statistically significant reductions observed in groups-4, 5, and 6. Histopathologically, group-2 displayed joint space narrowing, cartilage degeneration, and deep fissures. Groups-5 and 6 demonstrated significant joint space enlargement, articular cartilage tissue regeneration, and immunostaining patterns similar to those in group-1. Immunohistochemically, group-2 showed significant increases in IL-1β, TNF-α, MMP-13, and NOS-2 expression. However, all treatment groups exhibited reductions in these expression levels compared to group-2, with statistically significant decreases observed in groups-5 and 6 (P < 0.01).BA shows potential efficacy in reducing inflammation in experimental KOA model in rats. It may be a promising therapeutic agent for KOA, warranting further clinical studies for validation.

Evaluation of Citrus junos Essential Oil as a Promising Anti-aging Ingredient: In Vitro and Clinical Studies on Skin Health Improvement

Purpose: This study assessed Citrus junos essential oil (CJEO) as an anti-aging agent for improving skin health.Methods: CJEO was extracted from authenticated C. junos fruit peels, and its chemical composition was analyzed using a gas chromatographymass spectrometry (GC-MS). In vitro experiments were conducted on human epidermal keratinocytes (HEKs) and human dermal fibroblasts (HDFs) to evaluate CJEO’s impact on cell viability, anti-inflammatory activity, collagen synthesis, and collagen degradation. The expression of TNF-α, IL-1β, IL-6, NF-κB, MMP-1, MMP-3, COL1A1, and COL3A1 was quantified using RT-qPCR. A dermal patch test was performed on human participants to assess potential skin irritation, and clinical evaluations were conducted on 20 female participants to analyze CJEO’s effects on skin elasticity.Results: CJEO effectively inhibited nitric oxide production and significantly suppressed the expression of inflammatory markers, including TNF-α, IL-1β, IL-6, and NF-κB, as well as collagen degradation markers MMP-1 and MMP-3. Furthermore, CJEO treatment upregulated the expression of genes associated with collagen synthesis, specifically COL1A1 and COL3A1. Notably, significant improvements in skin elasticity were observed in the 20 female participants. The dermal patch test indicated excellent tolerance of the CJEO emulsion without causing irritation.Conclusions: CJEO exhibits promising potential as a bioactive ingredient in anti-aging cosmeceuticals. Its anti-inflammatory properties, ability to stimulate collagen synthesis, suggest efficacy in reducing skin aging and promoting overall skin health. Additionally, the dermal patch test confirms CJEO’s safety, supporting its suitability for use in cosmetic formulations. These findings endorse CJEO as an effective and safe component in anti-aging skincare products.

Kukoamine A protects mice against osteoarthritis by inhibiting chondrocyte inflammation and ferroptosis via SIRT1/GPX4 signaling pathway

AimsOsteoarthritis (OA) is one of the common chronic degenerative joint diseases, characterized by cartilage damage, subchondral bone changes, osteophyte formation, and synovitis. Kukoamine A (KuKA) is a bioactive compound isolated from Lycium chinense which is known as its anti-inflammatory activity. In this study, we detected the regulatory role of KuKA on OA both in vivo and in vitro. Materials and methodsMouse chondrocytes were cultured and mouse model of OA was established. Inflammatory mediator was measured by ELISA. The signaling pathway was tested by western blot analysis. Key findingsKuKA inhibited IL-1β-induced PGE2 and NO production and iNOS and COX-2 expression. IL-1β-induced MMP1 and MMP3 production was attenuated by KuKA. IL-1β-induced MDA, iron, and ROS were alleviated by KuKA. Meanwhile, GSH content, GPX4, Ferritin, SIRT1, Nrf2, and HO-1 expression were upregulated by KuKA. Furthermore, the inhibitory role of KuKA on IL-1β-induced inflammation, MMPs production, and ferroptosis were reversed by SIRT1 inhibitor. In vivo, KuKA could attenuate OA development in mouse model. KuKA markedly alleviated MMP1, MMP3, iNOS, and COX2 expression in OA mice. SignificanceIn conclusion, KuKA could inhibit OA development through suppressing chondrocyte inflammation and ferroptosis via SIRT1/GPX4 signaling pathway.

Transcription Factor TCF3 Promotes Macrophage-Mediated Inflammation and MMP Secretion in Abdominal Aortic Aneurysm by Regulating miR-143-5p /CCL20.

Damage to the abdominal aortic wall and the local inflammatory response are key factors resulting in abdominal aortic aneurysm (AAA) formation. During this process, macrophage polarization plays a key role. However, in AAA, the regulatory mechanism of macrophages is still unclear, and further research is needed. In this study, we found that the transcription factor TCF3 was expressed at low levels in AAA. We overexpressed TCF3 and found that TCF3 could inhibit MMP and inflammatory factor expression and promote M2 macrophage polarization, thereby inhibiting the progression of AAA. Knocking down TCF3 could promote M1 polarization and MMP and inflammatory factor expression. In addition, we found that TCF3 increased miR-143-5p expression through transcriptional activation of miR-143-5p , which further inhibited expression of the downstream chemokine CCL20 and promoted M2 macrophage polarization. Our research indicates that TCF3-mediated macrophage polarization plays a key regulatory role in AAA, complementing the role and mechanism of macrophages in the occurrence and development of AAA and providing a scientific basis for AAA treatment.

Brevilin A attenuates cartilage destruction in osteoarthritis mouse model by inhibiting inflammation and ferroptosis via SIRT1/Nrf2/GPX4 signaling pathway

Osteoarthritis (OA) is a serious orthopedic disease that affects people's quality of life. Although there are many treatment methods, the treatment effect is still not good. Brevilin A is a bioactive compound isolated from the medicinal herbCentipeda minima. The potential efficacy of brevilin A on OA was explored in this study. Mouse chondrocytes were isolated and stimulated by IL-1β and mouse OA model was induced by destabilization of the medial meniscus (DMM). The results demonstrated that brevilin A markedly inhibited IL-1β-induced MMP1 and MMP3 production. IL-1β-induced PGE2, NO, MDA, and iron production were alleviated by brevilin A. The production of GSH and the expression of SIRT1, Nrf2, HO-1, GPX4, and Ferritin were increased by brevilin A. Furthermore, the inhibition of brevilin A on IL-1β-induced inflammation and ferroptosis were prevented by SIRT1 inhibitor. In vivo, the results showed brevilin A markedly attenuated OA progression in DMM-induced mouse OA model. Also, brevilin A could alleviate MMP1, MMP3, iNOS, and COX2 expression in OA mice. In conclusion, brevilin A protected mice against OA via suppressing inflammatory response and ferroptosis by regulating SIRT1/Nrf2/GPX4 signaling.

Panax notoginseng saponins inhibits oxidative stress- induced human nucleus pulposus cell apoptosis and delays disc degeneration in vivo and in vitro

Ethnopharmacological relevancePanax notoginseng (Burk) F. H. Chen has been a popular traditional Chinese medicine with a long history of treating low back pain. Its main active ingredient, Panax notoginseng saponins (PNS), can be found in several Chinese patent medicines that are frequently used to treat blood stasis type low back pain. Intervertebral disc degeneration (IDD) is the most common cause of back pain, and the injection of PNS has been used to relieve IDD-induced back pain in clinical practice. Despite its effectiveness, the exact mechanisms of action for PNS injections remain unclear. Aim of the studyIDD as a consequence of aging involves apoptosis of nucleus pulposus (NP) cells and imbalanced degradation of extracellular matrix (ECM) induced by several factors including oxidative stress. We hypothesized that PNS may have a therapeutic effect on IDD via inhibiting apoptosis of NP cells and degradation of ECM under oxidative stress. Materials and methodsIn this study, network pharmacology was initially employed to predict the targets of PNS against IDD. Subsequently, commercial PNS was analyzed by high-performance liquid chromatography to confirm the ingredients for in vitro and in vivo experiments. In vitro experiments were conducted on human nucleus pulposus (HNP) cells, including CCK-8, RT-PCR, Western blot, immunofluorescence staining, autophagic flux detection, and TUNEL assay. In vivo experiments were also performed on rats with IDD of tail discs induced by annular fibrosus needle puncture, which involved MRI, HE staining, and immunohistochemistry. ResultsOur study demonstrated the theoretical targets of PNS against IDD, including Caspase 3, MMP13, Akt, and autophagy, based on network pharmacology. Subsequently, in vitro experiments revealed that PNS attenuated cellular apoptosis of NP by suppressing the expression of cleaved-caspase 3 and the ratio of Bax/Bcl-2 under H2O2 stimulation. Autophagy was also inhibited by PNS treatment, and the protective effect was abolished with rapamycin, an autophagy inducer, indicating that autophagy inhibition was involved in the protective effect of PNS on IDD. Furthermore, Akt/mTOR pathway activation was observed in HNP cells responding to H2O2 with PNS treatment, which played a role in autophagy downregulation. PNS was also shown to promote the expression of anabolic genes such as COL2A1 and ACAN while inhibiting the expression of catabolic gene MMP13 in HNP cells. In addition, the in vivo study revealed that PNS treatment could ameliorate IDD in a puncture-induced rat tail model. The development of IDD was significantly reduced, and there was decreased MMP13 expression, as well as increased COL2A1 protein expression in NP tissues. ConclusionOur study showed that PNS could protect HNP cells against apoptosis via autophagy inhibition and ameliorate disc degeneration in vivo, indicating its potential to be a therapeutic agent for IDD.

Genistein Suppresses IL-6 and MMP-13 to Attenuate Osteoarthritis in Obese Diabetic Mice.

Type 2 diabetes mellitus and osteoarthritis (OA) often present as comorbidities. We examined the role of plasma IL-6, chondrocyte MMP-13, and col10a expression in the development of OA in obese diabetic mice. We further investigated dietary genistein and exercise training as potential mitigators of OA. One hundred adult mice (50 females, 50 males) aged 6 weeks were randomized into 5 groups, including lean controls, obese diabetic controls, and obese diabetic mice treated with genistein, exercise training, and genistein plus exercise training. The obese diabetic state was induced by feeding the mice a high-fat, high-sugar diet. Genistein was incorporated into the diet at a concentration of 600 mg genistein/kg. Exercise training was performed on a treadmill and consisted of daily 30 min sessions at 12 m/min, 5 days/week for a 12-week period. After treatment, plasma was collected, and proximal tibias were removed for analysis. Plasma IL-6 and MMP-13 were elevated while col10a was reduced in obese diabetic mice in comparison to lean controls. Dietary genistein treatment reduced IL-6 and MMP-13 expression and increased col10a expression. Histological examination of articular cartilage showed reduced thickness of the uncalcified zones and proteoglycan content in the cartilage of diabetic mice in comparison to mice fed genistein. Exercise training had no significant effect. In conclusion, genistein (and not exercise training) attenuates OA by reducing IL-6 and MMP-13 expression in diabetic mice.

N-acetyltransferase 10 promotes the progression of oral squamous cell carcinoma through N4-acetylcytidine RNA acetylation of MMP1 mRNA.

The pathogenesis of oral squamous cell carcinoma (OSCC) remains unclear. Therefore, clarifying its pathogenesis and molecular-level development mechanism has become the focus of OSCC research. N-acetyltransferase 10 (NAT10) is a crucial enzyme involved in mRNA acetylation, regulating target gene expression and biological functions of various diseases through mediating N4-acetylcytidine (ac4C) acetylation. However, its role in OSCC progression is not well understood. In this study, we showed that NAT10 was significantly upregulated in OSCC tissues compared to normal oral tissues. Moreover, lentivirus-mediated NAT10 knockdown markedly suppressed cell proliferation, migration, and invasion in two OSCC cell lines (SCC-9 and SCC-15). Interestingly, MMP1 was found to be significantly upregulated in OSCC tissues and was a potential target of NAT10. N-acetyltransferase 10 knockdown significantly reduced both the total and ac4C acetylated levels of MMP1 mRNA and decreased its mRNA stability. Xenograft experiments further confirmed the inhibitory effect of NAT10 knockdown on the tumorigenesis and metastasis ability of OSCC cells and decreased MMP1 expression in vivo. Additionally, NAT10 knockdown impaired the proliferation, migration, and invasion abilities in OSCC cell lines in an MMP1-dependent manner. Our results suggest that NAT10 acts as an oncogene in OSCC, and targeting ac4C acetylation could be a promising therapeutic strategy for OSCC treatment.

Carboxyamidotriazole alleviates pannus formation and cartilage erosion in rats with adjuvant arthritis

Carboxyamidotriazole (CAI) was initially considered a non-cytotoxic anticancer agent. However, recently, pronounced anti-inflammatory properties of CAI have been reported. Rheumatoid arthritis (RA) is an autoimmune inflammatory disease characterized by aberrant activation of signaling pathways. Therefore, this study explored the therapeutic effects and potential mechanism of action of CAI on RA in the adjuvant arthritis (AA) model. The results showed that CAI reduced the severity of arthritis in AA rats as demonstrated by inhibited hind paw swelling, reduced body weight, and decreased infiltration of joint pathological inflammatory cells. Importantly, pathological scoring of new blood vessels and immunohistochemical assays revealed that CAI inhibited pannus formation. CAI decreased the expression of pro-angiogenic growth factors, such as vascular epidermal growth factor, basic fibroblast growth factor, and metalloproteinases (MMPs), namely, MMP-1 and MMP-3 in the synovium of AA rats. Furthermore, CAI significantly reduced the increased levels of phosphorylated p38, c-Jun N-terminal kinase (JNK)1/2, and extracellular signal-regulated kinase (ERK)1/2 proteins in AA rats. In addition, the proliferation of fibroblast-like synoviocytes (FLS) was downregulated by CAI both in vivo and in vitro. In conclusion, this investigation illustrates the therapeutic effect of CAI on synovitis and erosion of articular cartilage in RA. Furthermore, the mechanism might involve inhibition of aberrantly activated mitogen-activated protein kinase signaling, as well as a decrease in pro-angiogenic factors, MMP expression, and FLS proliferation.

Effect of Aloe Vera On Mmp-1 And Timp-1 Expression on Diabetic Wound Healing

Aim: The aim of this study is to investigate the healing aspect of aloe vera in diabetes mellitus, which inhibits wound healing.&#x0D; Methods: Diabetes model was created with streptozotocin. At the end of the 14-day experiment, blood glucose was measured from the tail vein of animals in all groups and blood was taken from the heart and sacrificed. Histopathology and immunohistochemical statistics and evaluation were performed.&#x0D; Results: Pycnosis and degeneration of epithelial cells were observed in diabetes groups. Leukocyte infiltration in the dermal papilla, degeneration of collagen fibers and an increase in the extracellular matrix were observed. It was observed that the epithelial layer in the aloe vera group was histologically close to the control group. It was observed that decreased inflammation in the dermal papilla and decreased in organized collagen fibers and vessel dilatation were observed. In the control group, MMP-1 and TIMP-1 expression were positive in the epidermis and dermis layers. In the diabetes group, weak expression of MMP-1 and TIMP-1 was observed in cells in the epidermis and dermis. The expression of MMP-1 and TIMP-1 in the surface epithelium in the aloe vera group was increased compared to the diabetes group.&#x0D; Conclusions: Aloe vera accelerated cell and extracellular matrix regeneration with its anti-oxidative activity