TIMP-1 Expression Research Articles

Intravenous Injection of Sodium Hyaluronate Diminishes Basal Inflammatory Gene Expression in Equine Skeletal Muscle.

Following strenuous exercise, skeletal muscle experiences an acute inflammatory state that initiates the repair process. Systemic hyaluronic acid (HA) is injected to horses routinely as a joint anti-inflammatory. To gain insight into the effects of HA on skeletal muscle, adult Thoroughbred geldings (n = 6) were injected with a commercial HA product weekly for 3 weeks prior to performing a submaximal exercise test. Gluteal muscle (GM) biopsies were obtained before and 1 h after exercise for gene expression analysis and HA localization. The results from RNA sequencing demonstrate differences in gene expression between non-injected controls (CON; n = 6) and HA horses. Prior to exercise, HA horses contained fewer (p < 0.05) transcripts associated with leukocyte activity and cytokine production than CON. The performance of exercise resulted in the upregulation (p < 0.05) of several cytokine genes and their signaling intermediates, indicating that HA does not suppress the normal inflammatory response to exercise. The transcript abundance for marker genes of neutrophils (NCF2) and macrophages (CD163) was greater (p < 0.05) post-exercise and was unaffected by HA injection. The anti-inflammatory effects of HA on muscle are indirect as no differences (p > 0.05) in the relative amount of the macromolecule was observed between the CON and HA fiber extracellular matrix (ECM). However, exercise tended (p = 0.10) to cause an increase in ECM size suggestive of muscle damage and remodeling. The finding was supported by the increased (p < 0.05) expression of CTGF, TGFβ1, MMP9, TIMP4 and Col4A1. Collectively, the results validate HA as an anti-inflammatory aid that does not disrupt the normal post-exercise muscle repair process.

Pilot Study on the Molecular Pathogenesis of Pyeloureteral Junction Obstruction: Underdevelopment or Fibrosis?

Background and Objectives: Congenital ureteral stenosis is one of the leading causes of impaired urinary drainage and subsequent dilatation of the urinary collecting system, known as hydronephrosis or ureterohydronephrosis. The mechanism that leads to obstruction is not clearly known. Multiple studies in rat models have shown increased angiotensin II and TGFβ levels in obstructed ureteral tissue. The aim of the study is to investigate the expression of fibrosis-related genes in obstructive and normal ureteral tissue. Material and Methods: It is a monocentric pilot study in which nineteen patients were selected prospectively. 17 patients underwent Hynes-Anderson pyeloplasty due to the PUJO; two patients underwent ureteroneocystostomy due to ureterovesical junction obstruction (UVJO); and six patients were chosen for the control group: five underwent nephrectomies due to the kidney tumor and one underwent upper pole heminephrectomy due to the duplex kidney with normal pyeloureteric junctions in all. Tissue RNA was chemically extracted after freezing the biopsy samples in liquid nitrogen, with cDNA synthesis performed immediately after nucleic acid isolation. qPCR was performed to evaluate the relative expression of Tgfb1, Mmp1, Timp1, Pai1, Ctgf, and Vegfa. Expression levels of the Gapdh and Gpi genes (geometric average) were used to calculate the relative expression of the investigated genes. Outliers were removed prior to calculating confidence intervals for the experimental groups, and a Wilcoxon rank-sum test was performed to determine the statistical significance of the differences. Results: Significant differences between healthy and stenotic tissue samples in Ctgf gene expression levels were observed, with the samples from afflicted tissue showing lower expression. No statistical difference in expression levels of Tgfb1, Timp1, Vegfa, Mmp1, and Pai1 was found. Conclusions: These findings suggest that tissue fibrosis, similar to other tissues and organs, is not the leading cause of stenosis, at least at the moment of surgery. Decreased CTGF expression is indicative of the developmental origin of obstruction.

Sublytic C5b-9 induces TIMP3 expression by glomerular mesangial cells via TRAF6-dependent KLF5 K63-linked ubiquitination in rat Thy-1 nephritis

Rat Thy-1 nephritis (Thy-1N) is an experimental model for studying human mesangioproliferative glomerulonephritis (MsPGN), and its pathological features are glomerular mesangial cell (GMC) proliferation and extracellular matrix (ECM) accumulation. Although we have confirmed that renal lesions of Thy-1N rats are sublytic C5b-9-dependent, and ECM accumulation is related to tissue inhibitor of matrix metalloproteinase (TIMP) inhibiting matrix metalloproteinase (MMP) activity, whether sublytic C5b-9 can induce TIMP production by GMC in Thy-1N rat and the underlying mechanism remains unclear. In the study, we proved that the expressions of TIMP3, krϋppel-like transcription factor 5 (KLF5) and tumor necrosis factor receptor-associated factor 6 (TRAF6) were simultaneously up-regulated both in the renal tissues of Thy-1N rats (in vivo) and in the GMC exposed to sublytic C5b-9 (in vitro). Further mechanism exploration discovered that KLF5 and TRAF6 as two upstream molecules could induce TIMP3 gene transcription through binding to the same region i.e., −1801nt to −1554nt (GGGGAGGGGC) and −228nt to −46nt (GCCCCGCCCC) of TIMP3 promoter. In the process, TRAF6 mediated KLF5 K63-linked ubiquitination at K99 and K100 enhancing KLF5 nuclear localization and binding to TIMP3 promoter, augmenting its gene activation. Furthermore, the experiments in vivo exhibited that silencing KLF5, TRAF6 or TIMP3 gene could markedly lessen renal KLF5 K63-linked ubiquitination or TIMP3 induction, ECM accumulation and other pathological changes of Thy-1N rats. Besides, the positive expressions of above-mentioned these proteins and ECM accumulation and their correlation in the renal tissues of MsPGN patients were also demonstrated. Overall, our findings implicate that KLF5 and TRAF6 play a promoting role in sublytic C5b-9-triggered TIMP3 gene transcription and expression, which might provide a novel mechanistic insight into rat Thy-1N and human MsPGN.

CircPTP4A2 Promotes Microglia Polarization in Cerebral Ischemic Stroke via miR-20b-5p/YTHDF1/TIMP2 Axis.

Activated microglia play dual roles in ischemic stroke (IS) according to its polarization states. Herein, we investigated the function of circPTP4A2 in regulating microglia polarization in IS. IS models were established by MACO/R and OGD/R treatment. TTC staining was employed to detect cerebral infarct size. Cell vitality was measured using CCK-8 assay. CD16 and CD206 levels were examined using flow cytometry. The interactions between circPTP4A2, miR-20b-5p, and YTHDF1 were analyzed by dual-luciferase reporter gene, RIP, or RNA pull-down assays. circPTP4A2 was upregulated in IS patients. circPTP4A2 knockdown alleviated MCAO/R-induced cerebral injury in mice. circPTP4A2 knockdown promoted microglia M2 polarization after OGD/R. circPTP4A2 promoted YTHDF1 expression by sponging miR-20b-5p. The promoting effect of circPTP4A2 knockdown on microglia M2 polarization was abrogated by miR-20b-5p inhibition. YTHDF1 activated the NF-κB pathway by increasing TIMP2 mRNA stability and expression. circPTP4A2 downregulation promoted microglia M2 polarization to inhibit IS development by regulating the miR-20b-5p/YTHDF1/TIMP2/NF-κB axis.

The role of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) Expression in pelvic organ prolapse: A literature review

Link of Video Abstract: https://youtu.be/Zv9b2Zj96yk Menopause is a physiological process as women get older. Urogenital syndrome, sexual difficulties, and pelvic organ prolapse (POP) are all common complaints among postmenopausal women, and these conditions can negatively impact their quality of life. There are still many unknowns regarding the pathophysiology and mechanisms of POP. Pelvic organ prolapse has been linked to the equilibrium of the extracellular matrix (ECM), which is controlled by matrix metalloproteinases and tissue inhibitors of metalloproteinases. Menopausal women experience a variety of symptoms due to hormonal changes, from urinary tract disturbance, vaginal atrophy, vaginal shortening, up to genital prolapse. Due to the collagen's diminished contractility, POP happens. Matrix metalloproteinase is responsible for breaking down collagen, but TIMP prevents MMP from doing its job. In women with POP caused by the breakdown of the collagen network, MMP-9 exhibits the greatest rise. In order to preserve the health of fibroblasts and collagen in postmenopausal women, increased expression of MMP-9 and decreased expression of TIMP-1 are necessary, which results in a decreased incidence of POP. The expression of MMP-9 in prolapse patients was significantly higher than control patients. In addition, TIMP-1 expression levels were significantly decreased in prolapse patients. Damage to the ECM's equilibrium is caused by increased MMP-9 expression and decreased expression of timp-1, which leads to clinical signs of pelvic organ prolapse.

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

Quantification of MMP-2 and TIMP-1 expressions in breast cancer

Quantification of MMP-2 and TIMP-1 expressions in breast cancer

Walnut Kernel Oil and Defatted Extracts Enhance Mesenchymal Stem Cell Stemness and Delay Senescence.

Decreased stemness and increased cellular senescence impair the ability of mesenchymal stem cells (MSCs) to renew themselves, change into different cell types, and contribute to regenerative medicine. There is an urgent need to discover new compounds that can boost MSCs' stemness and delay senescence. Therefore, this study aimed to investigate the impact of walnut kernel oil (WKO) and defatted (WKD) extracts on bone marrow (BM)-MSC stemness and senescence. Premature senescence and inflammation were induced in BM-MSCs using H2O2 and LPS, respectively. Phytochemical constituents of WKO and WKD extracts were detected by HPLC. The stemness (proliferation and migration), senescence-related markers (p53, p21, SIRT1, and AMPK), oxidative stress/antioxidant markers, inflammatory cytokines, and cell cycle of BM-MSCs were measured by MTT assay, qPCR, ELISA, and flow cytometry. WKO and WKD extracts improved rat BM-MSC stemness, as evidenced by (1) increased cell viability, (2) decreased apoptosis (low levels of Bax and caspase3 and high levels of Bcl2), (3) upregulated MMP9 and downregulated TIMP1 expression, and (4) cell cycle arrest in the G0/G1 phase and declined cell number in the S and G2/M phases. Additionally, WKO and WKD extracts reduced rat BM-MSC senescence, as indicated by (1) decreased p53 and p21 expression, (2) upregulated expression and levels of SIRT1 and AMPK, (3) reduced levels of ROS and improved antioxidant activity (higher activity of CAT, SOD, and GPx and upregulated expression of NrF2 and HO-1), and (4) declined levels of TNFα, IL1β, and NF-κB. When compared to the WKO extract, the WKD extract had a greater impact on the induction of stemness and reduction of senescence of BM-MSCs due to its stronger antioxidant activity, which could be attributed to its higher levels of flavonoids and phenolic compounds, as detected by HPLC analysis. WKO and WKD extracts enhance rat BM-MSC stemness and protect them from senescence, suggesting their potential use as enhancers to increase MSCs' therapeutic efficacy.

TMT-Based Proteomics Analysis of Senescent Nucleus Pulposus from Patients with Intervertebral Disc Degeneration.

Lower back pain, a leading cause of disability worldwide, is associated with intervertebral disc degeneration (IDD) in approximately 40% of cases. Although nucleus pulposus (NP) cell senescence is a major contributor to IDD, the underlying mechanisms remain unclear. We collected NP samples from IDD patients who had undergone spinal surgery. Healthy and senescent NP tissues (n = 3) were screened using the Pfirrmann grading system combined with immunohistochemistry, as well as hematoxylin and eosin, Safranin O, Alcian blue, and Masson staining. Differentially expressed proteins (DEPs) were identified using quantitative TMT-based proteomics technology. Bioinformatics analyses included gene ontology (GO) annotation, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and protein-protein interaction (PPI) analyses. In addition, immunofluorescence was used to verify protein expression. In total, 301 DEPs were identified in senescent NP tissues, including 92 upregulated and 209 downregulated proteins. In GO, DEPs were primarily associated with NF-kappaB transcription factor, extracellular regions, cellular protein metabolic processes, and post-translational protein modification. The enriched KEGG pathways included TGF-β, Wnt, RAP1, interleukin-17, extracellular matrix-receptor adhesion, and PI3K/Akt signaling pathways. PPI analysis demonstrated interactions between multiple proteins. Finally, immunofluorescence verified the expressions of MMP3, LUM, TIMP1, and CDC42 in senescent NP cells. Our study provides valuable insights into the mechanisms underlying senescent NP tissues in IDD patients. DEPs provide a basis for further investigation of the effects of senescent factors on IDD.

Novel CAF-identifiers via transcriptomic and protein level analysis in HNSC patients

Cancer-associated fibroblasts (CAFs), a prominent component of the tumor microenvironment, play an important role in tumor development, invasion, and drug resistance. The expression of distinct “CAF-markers” which separates CAFs from normal fibroblasts and epithelial cells, have traditionally been used to identify them. These commonly used CAF-markers have been reported to differ greatly across different CAF subpopulations, even within a cancer type. Using an unbiased -omic approach from public data and in-house RNAseq data from patient derived novel CAF cells, TIMP-1, SPARC, COL1A2, COL3A1 and COL1A1 were identified as potential CAF-markers by differential gene expression analysis using publicly available single cell sequencing data and in-house RNAseq data to distinguish CAF populations from tumor epithelia and normal oral fibroblasts. Experimental validation using qPCR and immunofluorescence revealed CAF-specific higher expression of TIMP-1 and COL1A2 as compared to other markers in 5 novel CAF cells, derived from patients of diverse gender, habits and different locations of head and neck squamous cell carcinoma (HNSC). Upon immunohistochemical (IHC) analysis of FFPE blocks however, COL1A2 showed better differential staining between tumor epithelia and tumor stroma. Similar data science driven approach utilizing single cell sequencing and RNAseq data from stabilized CAFs can be employed to identify CAF-markers in various cancers.

Inhibitory effects of Schisandrin C on collagen behavior in pulmonary fibrosis

Pulmonary fibrosis (PF) is a serious progressive fibrotic disease that is characterized by excessive accumulation of extracellular matrix (ECM), thus resulting in stiff lung tissues. Lysyl oxidase (LOX) is an enzyme involved in fibrosis by catalyzing collagen cross-linking. Studies found that the ingredients in schisandra ameliorated bleomycin (BLM)-induced PF, but it is unknown whether the anti-PF of schisandra is related to LOX. In this study, we established models of PF including a mouse model stimulated by BLM and a HFL1 cell model induced by transforming growth factor (TGF)-β1 to evaluate the inhibition effects of Schisandrin C (Sch C) on PF. We observed that Sch C treatment decreased pulmonary indexes compared to control group. Treatment of Sch C showed a significant reduction in the accumulation of ECM as evidenced by decreased expressions of α-SMA, FN, MMP2, MMP9, TIMP1 and collagen proteins such as Col 1A1, and Col 3A1. In addition, the expression of LOX in the lung tissue of mice after Sch C treatment was effectively decreased compared with the MOD group. The inhibition effects in vitro were consistent with those in vivo. Mechanistic studies revealed that Sch C significantly inhibited TGF-β1/Smad2/3 and TNF-α/JNK signaling pathways. In conclusion, our data demonstrated that Sch C significantly ameliorated PF in vivo and vitro, which may play an important role by reducing ECM deposition and inhibiting the production of LOX.

Early Continuous Passive Motion and Needle-knife Therapy Alleviate Knee Motor Dysfunction Effects After Internal Fixation of TPFs

Posttraumatic osteoarthritis (PTOA) is a common complication secondary to tibial plateau fractures (TPFs) after internal fixation. Early continuous passive motion (CPM) and needle-knife therapy are both alternative medicine treatments that have been beneficial to PTOA. CPM and needle-knife therapy are commonly applied separately in the treatment of orthopedic diseases and soft tissue injuries, but they both have limitations. In this study, we combined CPM and needleknife therapy to observe its efficacy on PTOA caused by postoperative TPFs and its possible mechanisms. A rabbit model of TPFs after internal fixation was established by surgical method. Rabbits were randomly divided into five groups: Normal control (NC) group, Sedentary (SED) group, CPM group, Needle-knife group, CPM + Needle-knife group. Range of motion (ROM), knee diameter and medial cartilaginous surface of the tibial plateau were observed after 4 weeks of treatment. Levels of inflammatory cytokines in the supernatants from knee joint fluid, including interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α, were determined by enzyme-linked immunosorbent assay (ELISA). The gross view of ligament tissues was detected by immunosorbent assay (ELISA). The gross view of ligament tissues was detected by histomorphology, including Hematoxylin and Eosin (H&amp;E) staining and Masson's trichrome staining. Other than that, the protein expression of matrix metalloproteinase (MMP)-13 and tissue inhibitors of metalloproteinase (TIMP)-1 were evaluated by immunohistochemical staining, respectively. Finally, the apoptosis of ligament cells was assessed through DAPI and TUNEL assay. Our data showed that ROM was reduced while knee diameter was increased in the SED group, but they were relieved by CPM and needle-knife—either singly or in combination treatment. CPM and needle-knife therapy also improved the effect of TPFs on other parameters, including the medial cartilaginous surface of the tibial plateau improvement, MMP-13 and TIMP-1 expression in the medial collateral ligament (MCL), and lateral collateral ligament (LCL), knee joint fluid IL-1β, IL-6, and TNF-α levels, and condition of ligament cells apoptosis. Our findings suggested that early CPM plus needle-knife therapy might be a useful and promising therapeutic method for PTOA.

Role of Syndecan-4 in the Inhibition of Articular Cartilage Degeneration in Osteoarthritis.

Despite its widespread existence, there are relatively few drugs that can inhibit the progression of osteoarthritis (OA). Syndecan-4 (SDC4) is a transmembrane heparan sulfate proteoglycan that modulates cellular interactions with the extracellular matrix. Upregulated SDC4 expression in articular cartilage chondrocytes correlates with OA progression. In the present study, we treated osteoarthritic cartilage with SDC4 to elucidate its role in the disease's pathology. In this in vitro study, we used real-time polymerase chain reaction (PCR) to investigate the effects of SDC4 on anabolic and catabolic factors in cultured chondrocytes. In the in vivo study, we investigated the effect of intra-articular injection of SDC4 into the knee joints of an OA mouse model. In vitro, SDC4 upregulated the expression of tissue inhibitor of metalloproteinase (TIMP)-3 and downregulated the expression of matrix metalloproteinase (MMP)-13 and disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-5 in chondrocytes. Injection of SDC4 into the knee joints of OA model mice prevented articular cartilage degeneration 6 and 8 weeks postoperatively. Immunohistochemical analysis 8 weeks after SDC4 injection into the knee joint revealed decreased ADAMTS-5 expression and increased TIMP-3 expression. The results of this study suggest that the treatment of osteoarthritic articular cartilage with SDC4 inhibits cartilage degeneration.

Abstract P1047: The Extracellular Matrix Metalloproteinase Inducer And The Membrane Type 1 Matrix Metalloproteinase In Marfan Syndrome Related Thoracic Aortic Aneurysm

Objectives: Marfan Syndrome (MFS) is a connective tissue disorder commonly associated with Thoracic Aortic Aneurysms (TAAs). TAAs occur because of abnormal remodeling of the aortic extracellular matrix (ECM). The balance between matrix metalloproteinases (MMPs) and the Tissue Inhibitors of MMPs (TIMPs), regulates these processes. The extracellular MMP inducer (EMMPRIN), is a widely expressed glycoprotein capable of stimulating the production of various MMPs, suggesting a direct role in driving pathology. The Membrane Type-1 MMP (MT1-MMP) can cause proteolytic shedding of EMMPRIN. Therefore, we investigated this relationship in MFS TAA tissue and plasma and tested the hypothesis that a unique proteolytic profile may exist. Methods: Levels of EMMPRIN, MT1-MMP, MMP-2, MMP-9, MMP-3, MMP-8, TIMP-1, and TIMP-2 were measured in aortic tissue and plasma from MFS patients with TAA and compared to healthy controls. Following modification of the abundance and location of MT1-MMP in isolated primary aortic fibroblasts, secreted EMMPRIN was measured in conditioned culture media. Results: Compared to controls, EMMPRIN is elevated in MFS TAA tissue and reduced in the plasma. Elevation of EMMPRIN abundance in tissue does not induce MMP-3, MMP-8, or TIMP-1 expression. MT1-MMP and TIMP-2 are elevated in MFS TAA tissue. MMP-2 and MMP-9 are reduced in MFS TAA tissue while increased in the plasma. In aortic fibroblasts we found that MT1-MMP must be internalized from the plasma membrane for EMMPRIN secretion. Conclusions: Our findings indicate that the pathological consequences of MFS cause the shedding of EMMPRIN and its secretion from cells to be blocked, which leads to higher EMPRINN abundance in MFS TAA tissue. This process depends upon the cellular location of MT1-MMP. Taken together, these observations suggest that internalization of MT1-MMP from the plasma membrane is required for the cellular secretion of EMMPRIN from aortic fibroblasts.

Novel Curcumin Analogue L6H4 in Treating Liver Fibrosis and Type 2 Diabetes.

The objective of this study was to evaluate the therapeutic efficacy of the curcumin analogue L6H4 in attenuating liver fibrosis and alleviating insulin resistance in streptozotocin-induced diabetic rats. Male Sprague-Dawley rats were fed a high-fat diet to induce insulin resistance, followed by streptozotocin injection to induce diabetes. The rats were then treated with L6H4 for eight weeks. Body weight, metabolic parameters, liver function, and liver histopathology were evaluated. Immunohistochemistry was performed to assess the expression of TGF-β1, TIMP-2, and MMP-2 in liver tissues. Statistical analysis was conducted using one-way ANOVA and Spearman rank correlation test. L6H4 treatment effectively reversed the weight gain associated with a high-fat diet and improved metabolic parameters in diabetic rats. Liver function markers, such as ALT and AST, were reduced after L6H4 treatment. Histological analysis showed improved liver morphology and reduced fibrosis in L6H4-treated rats. Electron microscopy revealed improved ultrastructural features of hepatocytes. Immunohistochemistry demonstrated downregulation of TGF-β1 and TIMP-2 expression and restoration of MMP-2 expression in the liver tissue of L6H4-treated rats. Correlation analysis showed a significant positive correlation between TGF-β1 and TIMP-2 expression. The findings suggest that L6H4 has therapeutic potential in attenuating liver fibrosis and alleviating insulin resistance in streptozotocin-induced diabetic rats. The hepatoprotective effect of L6H4 may be attributed to its anti-inflammatory properties and its ability to target molecules involved in fibrosis. Further research is warranted to explore the potential of L6H4 as a treatment option for nonalcoholic fatty liver disease and type 2 diabetes.

Ursolic Acid Ameliorated Neuronal Damage by Restoring Microglia-Activated MMP/TIMP Imbalance in vitro.

The oxygen and glucose deprivation-reoxygenation (OGDR) model is widely used to evaluate ischemic stroke and cerebral ischemia-reperfusion (I/R) injury in vitro. Excessively activated microglia produce pro-inflammatory mediators such as matrix metalloproteinases [MMPs] and their specific inhibitors, tissue inhibitors of metalloproteinases [TIMPs], causing neuronal damage. Ursolic acid (UA) acts as a neuroprotective agent in the rat middle cerebral artery occlusion/reperfusion (MCAO/R) model keeping the MMP/TIMP balance with underlying mechanisms unclear. Our study used OGDR model to determine whether and how UA reduces neuronal damage by reversing MMP/TIMP imbalance caused by microglia in I/R injury in vitro. SH-SY5Y cells were first cultured with 95% N2 and 5% CO2 and then cultivated regularly for OGDR model. Cell viability was tested for a proper UA dose. We established a co-culture system with SH-SY5Y cells and microglia-conditioned medium (MCM) stimulated by lipopolysaccharide (LPS) and interferon-gamma (IFNγ). MMP9 and TIMP1 levels were measured with ELISA assay to confirm the UA effect. We added recombinant MMP9 (rMMP9) and TIMP1 neutralizing antibody (anti-TIMP1) for reconfirmation. Transmission electron microscopy was used to observe cell morphology, and flow cytometry and Annexin V-FITC and PI labeling for apoptotic conditions. We further measured the calcium fluorescence intensity in SH-SY5Y cells. The MCM significantly reduced cell viability of SH-SY5Y cells after OGDR (p<0.01), which was restored by UA (0.25 µM) (p<0.05), whereas lactate dehydrogenase activity, intraneuronal Ca2+ concentration, and apoptosis-related indexes were showed significant improvement after UA treatment (p<0.01). UA corrected the MMP/TIMP imbalance by decreasing MMP9 expression and increasing TIMP1 expression in the co-culture system (p<0.01) and the effects of UA on SH-SY5Y cells were mitigated by the administration of rMMP9 and anti-TIMP1 (p<0.01). We demonstrated that UA inhibited microglia-induced neuronal cell death in an OGDR model of ischemic reperfusion injury by stabilizing the MMP9/TIMP1 imbalance.

Scoparone attenuates cholestatic liver injury through regulating hepatic bile acid metabolism, ameliorating periductal fibrosis and inhibiting inflammatory response

BackgroundCholestatic liver disease (CLD) encompasses a broad spectrum of etiologically heterogeneous hepatobiliary disorders with common features, including ductal reaction, periductal fibrosis, and inflammatory response. Scoparone (SCO) is a bioactive component of the herbal medicine Artemisia capillaris Thunb that is used clinically for the treatment of hepatic dysfunction, cholestasis and jaundice. AimTo investigate the therapeutic effect of SCO against 3,5-diethoxycarbonyl- 1,4- dihydrocollidine (DDC) induced CLD. Materials and methodsIn the present study, two independent experiments were conducted to comprehensively analyze the hepatoprotective and hepatotoxicity effects of SCO against DDC diet-induced CLD. First, three doses of SCO (5, 15, and 45 mg/kg) were injected intraperitoneal daily for 14 days in DDC diet-induced CLD for the pharmacological research. Then, a high dose of SCO (45 mg/kg) was used for 28 days in negative and CLD model mice to evaluate the therapeutic potential of SCO. ResultsDDC-induced CLD mice exhibited hepatomegaly and weight loss with hepatic pathological features including hepatocyte and bile duct damage, bile acid metabolism abnormalities, peribiliary fibrosis, and inflammatory factor infiltration. Treatment with three doses of SCO (5, 15 and 45 mg/kg) produced dose-dependent hepatoprotective effects, attenuated fine bile duct responses, ameliorated peribiliary fibrosis and suppressed intrahepatic inflammatory responses. SCO inhibited the expression of Cyp2b10, a bile acid phase I metabolizing enzyme, and Sult2a1, a phase II metabolizing enzyme, in DDC diet-induced CLD mice. Meanwhile, SCO inhibited the expression of Tgfb1, aSMA, Timp1 and Col1a1. SCO suppressed liver inflammation by reducing the infiltration of myeloid cells and lymphocytes. SCO suppressed DDC-induced lymphocyte infiltration. In addition, normal control mice treated with SCO showed no significant changes in liver injury, bile duct reaction, peribiliary fibrosis and inflammatory infiltration. ConclusionSCO exerts a pharmacological effect on the reprogramming of hepatic bile acid metabolism, hepatic fibrosis and inflammatory response, thus having potential for the treatment of CLD.

Critical Evaluation of Transcripts and Long Noncoding RNA Expression Levels in Prostate Cancer Following Radical Prostatectomy

Introduction: The clinical course of prostate cancer (PCa) is highly variable, ranging from indolent behavior to rapid metastatic progression. The Gleason score is widely accepted as the primary histologic assessment tool with significant prognostic value. However, additional biomarkers are required to better stratify patients, particularly those at intermediate risk. Methods: In this study, we analyzed the expression of 86 cancer hallmark genes in 171 patients with PCa who underwent radical prostatectomy and focused on the outcome of the 137 patients with postoperative R0-PSA0 status. Results: Low expression of the IGF1 and SRD52A, and high expression of TIMP2, PLAUR, S100A2, and CANX genes were associated with biochemical recurrence (BR), defined as an increase of prostate-specific antigen above 0.2 ng/mL. Furthermore, the analysis of the expression of 462 noncoding RNAs (ncRNA) in a sub-cohort of 39 patients with Gleason score 7 tumors revealed that high levels of expression of the ncRNAs LINC00624, LINC00593, LINC00482, and cd27-AS1 were significantly associated with BR. Our findings provide further evidence for tumor-promoting roles of ncRNAs in PCa patients at intermediate risk. The strong correlation between expression of LINC00624 and KRT8 gene, encoding a well-known cell surface protein present in PCa, further supports a potential contribution of this ncRNA to PCa progression. Conclusion: While larger and further studies are needed to define the role of these genes/ncRNA in PCa, our findings pave the way toward the identification of a subgroup of patients at intermediate risk who may benefit from adjuvant treatments and new therapeutic agents.

The effect of laser and dynamic quadripolar radiofrequency therapies on the expression of collagenogenesis proteins in the vaginal epithelium and tissue elasticity in women with genital prolapse

Background. Genital prolapse dramatically affects the quality of life of patients. Various sources show that the relapse rate after surgical treatment is 30 to 60%.&#x0D; Aim. To determine and compare the expression levels of collagen metabolism proteins before and after exposure to laser and radio frequency in the postoperative period in postmenopausal patients and to compare ultrasound elastography data to justify using these exposure methods as a rehabilitation program.&#x0D; Materials and methods. Histological specimens were collected from 54 patients with postmenopausal POP-Q grade 3 genital prolapse, and an ultrasound examination was made before surgery and after rehabilitation therapy. A real-time polymerase chain reaction method was used to assess the expression level of type I collagen, matrix metalloproteinases (MMP-2, MMP-9), and tissue inhibitors of metalloproteinases (TIMP-1, TIMP-2). Ultrasound examination was performed using the technology of compression elastography with the analysis of the color scale and the coefficient of tissue deformation.&#x0D; Results. Data were obtained on the increase in the expression of TIMP-1 by 7 times and TIMP-2 by 4 times in the radiofrequency therapy group compared to the group before treatment, as well as a 64-fold decrease in TIMP-2 expression in the vaginal tissue of women in the group after laser therapy compared to the group before treatment. A significant increase in the expression of TIMP-1 and TIMP-2 genes in vaginal tissue after radiofrequency exposure and a decrease in TIMP-2 after laser exposure demonstrate the compensatory effect of biological markers in response to energy therapy, as well as the remodeling effect of these methods. According to ultrasound, tissue elasticity in patients increased after laser and radiofrequency exposure.&#x0D; Conclusion. The results support using radiofrequency and laser therapy in the postoperative period as rehabilitation methods.

Significance of Tissue Inhibitors of Matrix Metalloproteinases Expression in Pathogenesis and Differential Diagnosis of Periodontal Pathology

Background. The balance of metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) is crucial for the manifestation and progression of periodontal diseases and is one of the promising areas of scientific research in terms of developing methods for selective inhibition of MMPs. Aims to determine the significance of TIMP1 and TIMP2 expression parameters in gingival biopsies to identify the type of periodontitis at the stage of disease manifestation. Methods. A prospective study with parallel inclusion of patients with different forms of periodontitis in the compared groups was performed. The object of this study were patients with various forms of periodontitis. Pathological examination with morphometric analysis of TIMPs expression and subsequent statistical data analysis was performed using AperioImageScope v12.4.0.5043, Statistica 10.0, MedCalc19.6. Results. The study included 67 patients with aggressive (AgP, Grade C, n = 19), chronic simplex (CSP, Grade В, n = 10) and chronic complex (CCP, in the presence of occlusal trauma, Grade В, n = 38) periodontitis and 15 conditionally healthy patients (control group). The expression of TIMP1 and TIMP2 was detected with variable intensity both in the epithelium and in the stroma of the gingiva, and was significantly higher in periodontitis groups compared with the control group (MeTIMP1/2 = 32%/70%). Parameters of TIMP1 and TIMP2 expression had no significant differences in the groups with AgP (MeTIMP1/2 = 84%/98%) and CCP (MeTIMP1/2 = 83%/94%), except for higher parameters of the positivity of epithelial expression of TIMP2 (U = 61 372; p 0.05) and lower levels of intensity of its expression in AgP (MeAgP/CCP=180/171; U = 56 491; p 0.001). Correlation analysis revealed an inverse relationship of TIMPs expression parameters with those MMPs, including those most significant for the development of AgP MMP1 ( = 0.40), MMP8 ( = 0.34), and MMP14 ( = 0.24). ROC analysis established acceptable informativeness of all studied parameters of total expression and positivity of epithelial expression of TIMP1, as well as positivity of total expression and intensity of epithelial expression of TIMP2 to distinguish between aggressive and chronic forms of periodontitis at the stage of disease manifestation. Conclusion. Our results show an increase in the expression of TIMP1 and TIMP2 in various forms of periodontitis and an inverse relationship with the MMPs inhibited by them. This complements both the fundamental knowledge of the development and progression of periodontal pathology and may have applied significance in terms of using the studied TIMP1 and TIMP2 expression criteria to establish the aggressive course of periodontitis already at the stage of disease manifestation, which will allow to individualize treatment, prevent or slow down tooth loss in order to preserve and/or improve the quality of life of this patient group.