Tissue Inhibitors Of Matrix Metalloproteinases Research Articles

Physiological Properties, Functions, and Trends in the Matrix Metalloproteinase Inhibitors in Inflammation-Mediated Human Diseases.

Matrix metalloproteinases (MMPs), also known as metalloproteinases, are enzymes that degrade proteins and require the presence of active metal atoms. There are more than 20 types of MMPs, and they promote cell migration through the proteolytic degradation of the extracellular basement. MMPs are upregulated in cancers and inflamed regions. MMPs have three conservation regions: pro-MMP, catalysis, and hemopexin. Through these domains, MMPs cleave matrixes and cell-cell barriers. Consequently, MMPs cleave the whole extracellular matrix (ECM). In other words, they decompose most of the components related to the ECM, in their roles as key enzymes in cellular and pathophysiological events in the body. Zn2+-containing endo-type peptidases directly degrade and remodel the ECM region in the progression of various diseases. MMPs are frequently found in abnormal disease status of inflammatory responses, periodontal lesion, inflammatory pulmonary lesion, arteriosclerotic smooth muscles, arthritis, and tumor metastasis and invasion. They are also known to participate in aging processes-such as wrinkle formation-by destroying collagen in the dermis. In particular, the onset of diseases via the MMP-dependent inflammatory response is caused by the breakdown of proteins in the ECM and the basement membranous region, which are the supporting structures of cells. This review describes the developments in the research examining the general and selective inhibitors for MMP associated with various human diseases over the past 20 years in terms of structure remodeling, substrate-recognizing specificities, and pharmacological applicability. Among two similar types of MMPs, MMP-2 is known as gelatinase-A with a 72 kDa, while MMP-9 is termed gelatinase-B with a 92 kDa. Both of these play a key role in this action. Therefore, both enzymatic expression levels coincide during the onset and progression of diseases. Endogenous tissue inhibitors of matrix metalloproteinases (TIMPs) are highly specific for each MMP inhibitor type. The intrinsic factors regulate various MMP types by inhibiting the onset of various diseases mediated by MMP-dependent or independent inflammatory responses. The MMP- 9 and MMP-2 enzyme activity related to the prognosis of diseases associated with the inflammatory response are selectively inhibited by TIMP1 and TIMP2, respectively. The major pathogenesis of MMP-mediated diseases is related to the proliferation of inflammatory cells in various human tissues, which indicates their potential to diagnose or treat these diseases. The discovery of a substance that inhibits MMPs would be very important for preventing and treating various MMP-dependent diseases. Considerable research has examined MMP inhibitors, but most of these have been synthetic compounds. Research using natural products as MMP inhibitors has only recently become a subject of interest. This review intends to discuss recent research trends regarding the physiological properties, functions, and therapeutic agents related to MMPs.

IRISIN STIMULATES ANABOLISM AND MATRIX SYNTHESIS IN HUMAN NUCLEUS PULPOSUS CELLS IN VITRO: NEW INSIGHTS INTO A CROSS-TALK BETWEEN THE MUSCLE AND THE INTERVERTEBRAL DISC

This study aimed to investigate the effect of irisin on human nucleus pulposus cells (hNPCs) in vitro. Our hypothesis was that irisin would improve hNPC metabolism and proliferation.hNPCs were isolated from intervertebral discs and cultured in alginate beads. hNPCs were exposed to phosphate-buffered saline (PBS) or recombinant irisin (r-irisin) at 5, 10 and 25 ng/mL (n=4). Each experiment was performed in triplicate. Cell proliferation was assessed with trypan blue staining-automated cell counting and PicoGreen assay. Glycosaminoglycan (GAG) content was measured using the DMMB assay. Metabolic activity was assessed with the MTT assay and the Griess Reagent System. Gene expression of collagen type II (COL2), matrix metalloproteinase (MMP)-13, tissue inhibitor of matrix metalloproteinase (TIMP)-1 and −3, aggrecan, interleukin (IL)-1β, a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-5 was measured by RT-PCR. MTT assay and ADAMTS-5, COL2, TIMP-1 and IL-1β gene expression were evaluated following incubation with 5, 10 and 25 ng/mL r-irisin for 24 hours and subsequent culture with 10 ng/ml IL-1β and vice versa (incubation for 24 hours with IL-1β and subsequent culture with r-irisin).Irisin increased hNPC proliferation (p<0.001), metabolic activity (p<0.05), GAG content (p<0.01), as well as COL2 (p<0.01), aggrecan (p<0.05), TIMP-1 and −3 (p<0.01) gene expression, while decreasing MMP-13 (p<0.05) and IL-1β (p<0.001) mRNA levels. r-irisin pretreatment of hNPCs cultured in pro-inflammatory conditions resulted in a rescue of metabolic activity (p<0.001) and a decrease of IL-1β (p<0.05) levels. Similarly, incubation of hNPCs with IL-1β and subsequent exposure to r-irisin increased hNPC metabolic activity (p<0.001), COL2 gene expression (p<0.05) and decreased IL-1β (p<0.05) and ADAMTS-5 levels (p<0.01).Irisin stimulates hNPC proliferation, metabolic activity, and anabolism by reducing IL-1β and catabolic enzyme expression while promoting matrix synthesis.

Evaluation of Matrix Metalloproteases by Artificial Intelligence Techniques in Negative Biopsies as New Diagnostic Strategy in Prostate Cancer.

Usually, after an abnormal level of serum prostate-specific antigen (PSA) or digital rectal exam, men undergo a prostate needle biopsy. However, the traditional sextant technique misses 15-46% of cancers. At present, there are problems regarding disease diagnosis/prognosis, especially in patients' classification, because the information to be handled is complex and challenging to process. Matrix metalloproteases (MMPs) have high expression by prostate cancer (PCa) compared with benign prostate tissues. To assess the possible contribution to the diagnosis of PCa, we evaluated the expression of several MMPs in prostate tissues before and after PCa diagnosis using machine learning, classifiers, and supervised algorithms. A retrospective study was conducted on 29 patients diagnosed with PCa with previous benign needle biopsies, 45 patients with benign prostatic hyperplasia (BHP), and 18 patients with high-grade prostatic intraepithelial neoplasia (HGPIN). An immunohistochemical study was performed on tissue samples from tumor and non-tumor areas using specific antibodies against MMP -2, 9, 11, and 13, and the tissue inhibitor of MMPs -3 (TIMP-3), and the protein expression by different cell types was analyzed to which several automatic learning techniques have been applied. Compared with BHP or HGPIN specimens, epithelial cells (ECs) and fibroblasts from benign prostate biopsies before the diagnosis of PCa showed a significantly higher expression of MMPs and TIMP-3. Machine learning techniques provide a differentiable classification between these patients, with greater than 95% accuracy, considering ECs, being slightly lower when considering fibroblasts. In addition, evolutionary changes were found in paired tissues from benign biopsy to prostatectomy specimens in the same patient. Thus, ECs from the tumor zone from prostatectomy showed higher expressions of MMPs and TIMP-3 compared to ECs of the corresponding zone from the benign biopsy. Similar differences were found for expressions of MMP-9 and TIMP-3, between fibroblasts from these zones. The classifiers have determined that patients with benign prostate biopsies before the diagnosis of PCa showed a high MMPs/TIMP-3 expression by ECs, so in the zone without future cancer development as in the zone with future tumor, compared with biopsy samples from patients with BPH or HGPIN. Expression of MMP -2, 9, 11, and 13, and TIMP-3 phenotypically define ECs associated with future tumor development. Also, the results suggest that MMPs/TIMPs expression in biopsy tissues may reflect evolutionary changes from prostate benign tissues to PCa. Thus, these findings in combination with other parameters might contribute to improving the suspicion of PCa diagnosis.

Activity of matrix metalloproteinase type 9 and its relationship with calcification of the thoracic aorta in patients with resistant hypertension: cross-sectional comparative study

Background. The extracellular matrix of vessels is degraded under the influence of matrix metalloproteinases (MMPs). MMP type 9 is of particular interest in arterial hypertension (AH) since its activity is regulated by a tissue inhibitor of matrix metalloproteinase type 1 (TIMP1). The balance of their ratio can shift in either direction depending on the disease stage and provoke vascular wall calcification.
 Objective. This cross-sectional comparative study investigated the level of MMP9 and its type 1 tissue inhibitor and their relationship with the severity of thoracic aorta calcification in patients with resistant hypertension, depending on antihypertensive therapy effectiveness.
 Material and methods. The study included 92 patients with resistant hypertension. All patients underwent 24-hour blood (BP) pressure monitoring (ABPM), and had their index of the thoracic aorta (ТАС) and the plasma levels of MMP9 and TIMP1 assessed.
 Results. Patients were divided into groups based on whether they achieved the BP target level according to the results of ABPM: controlled (n=44) and uncontrolled (n=48) resistant AH. In the second group, most estimated parameters of ABPM were higher, and the variants of the daily BP profile for non-dipper and night-peaker systolic blood pressure (SBP) were more common than in the first group. In the second group, a more pronounced increase in TAC, TIMP1, and TIMP1/MMP9 levels was revealed. Evaluation of the relationship between the studied parameters revealed a positive relationship between TAC and age (r=0.683, p 0.00001), hypertension duration (r=0.610, p 0.00001) and a negative relationship with regular antihypertensive therapy duration (r=0.822, p 0.00001). ТАC was higher in individuals with higher values of SBP (r=0.513, р 0.00001), pulse BP (r=0.805, р 0.00001), rate of morning rise in BP (r=0.678, р 0.00001) and insufficient decrease in SBP at night (r=-0.822, р 0.00001). There was a positive correlation of moderate strength TAC with the level of low-density lipoproteins (r=0.490, p=0.0002), TIMP1 (r=0.344, p=0.005) and the TIMP1/MMP9 ratio (r=0.481, p 0.00001).
 Conclusion. In patients with resistant hypertension, an increase in the TAC was revealed, which has a statistically significant direct correlation with the value of pulse BP, the morning rise in BP, and an insufficient decrease in SBP at night. An imbalance in the TIMP1/MMP9 ratio, rather than an isolated increase in MMP9, can indicate the intensity of the formation and degradation of the extracellular matrix components in patients with resistant AH, which triggers the vascular wall calcification process.

Abstract 4456: Inducible nitric oxide synthase (iNOS) and cycloxygenase-2 (COX2) inhibition reprogram the tumor microenvironment and suppress tumor growth in hepatocellular carcinoma

Abstract Hepatocellular carcinoma is a common cancer worldwide and a leading cause of cancer-related death. Chronic inflammation, accumulation of genetic changes and alternations of the liver microenvironment are critical during the process of carcinogenesis and development of HCC. Inducible nitric oxide synthase (iNOS or NOS2)-derived NO and cyclooxygenase-2 (COX2) are important parts of the neoplastic inflammatory environment. Continuous exposure to moderate to high concentrations of NO, produced by inducible NO synthase (iNOS) and COX2, promotes neoplastic transformation, chemotherapeutic resistance, enhanced cell proliferation, increased inflammation and immune resistance. Our previous study reported that iNOS promoted stem-like characteristics and activation of liver cancer stem cells through Notch signaling pathway. iNOS activity in both the liver cancer stem cells and the microenvironment contributes to tumor progression in mice, suggesting that targeting iNOS in the entire tumor could have therapeutic benefit in HCC. iNOS-derived NO and COX2-derived prostaglandin E2 (PGE2) were shown to promote feed-forward iNOS/COX2 crosstalk because NO induced COX2 and PGE2 induced NOS2. Our research showed that iNOS inhibition with the iNOS inhibitor (1400W) and COX2 inhibitor (Celebrex) diminished HCC tumor growth. The cytokine analysis revealed considerably lower levels of inflammatory cytokines including, IL-6, tissue inhibitor of matrix metalloproteinase-1 (TIMP1), macrophage inflammatory protein-3alpha (MIP-3alpha), IL-1B, CC chemokine subfamily of eosinophil chemotactic proteins eotaxins (CCL11), M-CSF, thymus and activation-regulated chemokine (TARC, CCL17) and leukemia inhibitory factor (LIF) in HCC with iNOS/COX2 inhibition. iNOS/COX2 levels influence the polarization and spatial location of lymphoid cells including CD8+ T cells. Targeting iNOS/COX2 blockade improved CD8+ T cell penetration into the tumor core. We also found that iNOS/COX2 blockade result in more CD4+ T helper cells and CD8+ tumor infiltrating lymphocyte (TIL), but reduce the number of exhausted CD4+ T cells and CD8+ T cells (PD1-high, Lag3+, CD39+ Tex). The results suggesting that iNOS/COX2 inhibitor therapy may alleviate HCC growth by promoting a anti-tumorigenic TME, modifying lymphoid spatial localization and gene expression phenotypes and decreasing T-cell exhaustion. Citation Format: Ronghua Wang, Christof Kaltenmeier, Ruiqi Yang, Tony Haykal, Hamza Yazdani, Celine Tohme, David Geller, Samer Tohme, Timothy Billiar. Inducible nitric oxide synthase (iNOS) and cycloxygenase-2 (COX2) inhibition reprogram the tumor microenvironment and suppress tumor growth in hepatocellular carcinoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4456.

Ursolic acid promotes microglial polarization toward the M2 phenotype via PPARγ regulation of MMP2 transcription

Microglia, which are the primary inflammatory cells of the brain, can undergo phenotypic switching between M1 and M2 polarization, which have opposing effects on inflammation. Peroxisome proliferator-activated receptor gamma (PPARγ) is a member of the nuclear receptor family of ligand-inducible transcription factors, and PPARγ is known to regulate M2 macrophage polarization. Previous studies have shown that the natural pentacyclic triterpenoid ursolic acid (3β-hydroxy-urs-12-en-28-oic acid; UA) influences microglial activation. Additionally, UA increases tissue inhibitor matrix metalloproteinase 1 (TIMP1), while greatly reducing the release of matrix metalloproteinase 2 (MMP2) and MMP9 in a PPARγ-dependent manner. Here, we examined the anti-inflammatory properties of UA by observing how well it promotes the phenotypic transition of lipopolysaccharide (LPS) and interferon gamma (IFNγ)-activated BV2 microglia from M1 to M2 polarization. To determine if PPARγ is involved in the underlying molecular pathway, we treated rats with UA and the PPARγ inhibitor BADGE. We also investigated the mechanisms by which PPARγ controls transcription from the MMP2 promoter. The in-vitro experiments showed that UA shifted LPS/IFNγ-activated BV2 microglia from the M1 to the M2 phenotype, which was associated with a reduction in the neurotoxic factors MMP2 and MMP9, and an increase in the anti-inflammatory factor TIMP1. Co-treatment with increased MMP2 and MMP9 synthesis while decreasing TIMP1 release, indicating that UA has anti-inflammatory effects on LPS/IFNγ-activated BV2 cells via activation of PPARγ. Next, we found that PPARγ directly influences MMP2 transcriptional activity by identifying the crucial peroxisome proliferator response element (PPRE) among five potential PPREs in the MMP2 promoter. These results suggest that UA has a protective anti-inflammatory effect against neuroinflammatory toxicity, which is exerted by direct activation of PPARγ and selectively modulates microglial polarization and suppresses MMP2 formation.

Assessing clinical and molecular outcomes of prophylactic thermal pulsation therapy on ocular surface health following refractive surgery.

To study ocular surface signs, symptoms, and tear film composition following prophylactic thermal pulsation therapy (TPT) prior to refractive surgery, and to compare these outcomes with those who underwent TPT after refractive surgery. Patients with mild-to-moderate evaporative dry eye disease (DED) and/or meibomian gland dysfunction (MGD) undergoing refractive surgery were included. Group 1 patients received TPT (LipiFlow) prior to laser-assisted in situ keratomileusis (LASIK; n = 32, 64 eyes), and Group 2 patients received TPT three months after LASIK (n = 27, 52 eyes). Ocular Surface Disease Index (OSDI) score, Schirmer's test (ST1, ST2), Tear Breakup Time (TBUT), meibography, and tear fluid were obtained preoperatively and at three months postoperatively in Groups 1 and 2. Additional postoperative evaluation was performed three months after TPT in Group 2. Tear soluble factor profile was measured by multiplex enzyme-linked immunosorbent assay (ELISA) using flow cytometry. Postoperative OSDI score was significantly lower and TBUT was significantly higher when compared with matched preoperative values of Group 1 participants. On the other hand, the postoperative OSDI score was significantly higher and TBUT significantly lower when compared with matched preoperative values of Group 2 participants. TPT significantly reduced the postoperative elevation in OSDI and significantly reduced the postoperative reduction in TBUT in Group 2 participants. Tear Matrix metalloproteinase-9/ Tissue inhibitor matrix metalloproteinase 1 (MMP-9/TIMP1) ratio was significantly higher, postoperatively, when compared with matched preoperative levels in Group 2. However, MMP9/TIMP1 ratio remained unaltered in Group 1 participants. TPT prior to refractive surgery improved postsurgical ocular surface signs and symptoms and reduced tear inflammatory factors, thereby suggesting the plausibility of reduced post-refractive surgery DED in patients.

Urinary levels of potentially toxic elements (PTEs) in female beauticians and their association with urinary biomarkers of oxidative stress/inflammation and kidney injury

The present research was aimed to assess the urinary levels of potentially toxic elements (PTEs) in female beauticians and its correlation with oxidative stress/inflammation and kidney injury. To this end, the urine samples were collected from 50 female beauticians from beauty salons (exposed group) and 35 housewives (control group), and then, the level of PTEs was determined. The mean levels of the sum of urinary PTEs (∑PTEs) biomarkers in before and after exposure and control group were 83.55, 114.27 and 13.61 μg/L, respectively. Results also showed that the urinary level of PTEs biomarkers is significantly higher in women occupationally exposed to cosmetics compared to control group. The urinary levels of arsenic (As), cadmium (Cd), lead (Pb), and chromium (Cr) biomarkers have high correlation coefficients with early oxidative stress effects such as 8-Hydroxyguanosine (8-OHdG), 8-isoprostane and Malondialdehyde (MDA). Moreover, As and Cd biomarker levels were positively and significantly associated with kidney damages such as urinary kidney injury molecule-1 (uKIM-1) and tissue inhibitor matrix metalloproteinase 1 (uTIMP-1) (P < 0.01). Therefore, women who working in beauty salons can probably be categorized as high – exposure and high-risk workers in terms of DNA oxidative and kidney damages.

Comparison of MMP-2 and TIMP-2 expressions in yak testes at different ages.

This study aimed to investigate the distribution and expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) in yak testes. The testes of healthy yaks at different ages: newborn [3 days], young [1 year], adult [4 years], and old [9 years] were collected for microscopic analyses using hematoxylin and eosin staining, immunohistochemistry and immunofluorescence, as well as western blot to compare the expression of MMP-2 and TIMP-2. Furthermore, the levels of MMP-2mRNA and TIMP-2mRNA was detected by real-time quantitative polymerase chain reaction (qPCR). The results of immunohistochemistry and immunofluorescence demonstrated that MMP-2 and TIMP-2 were mainly located in gonocytes of newborn, Sertoli cells of young, spermatozoa of adult and Leydig cells of old. The protein levels of MMP-2 and TIMP-2 exhibited a downward from newborn to adult, but increased again in old yaks. The analysis of qPCR showed that MMP-2 was higher in young compared with newborn or adult(**P<0.01), but a lower expression was detected in adult compared with old yak testicular tissues (*P<0.05). Compared with adults, TIMP-2 was significantly higher in newborn and young yaks (**P<0.01), and slightly higher in old yaks (*P<0.05). Hence, The location of MMP-2 and TIMP-2 in gonocytes were associated with the development of newborn yak testes. The expression of MMP-2 and TIMP-2 in Sertoli cells at young and adult yaks suggested that they provided a clue for the regulation of spermatogenesis. The positive labeling of MMP-2 and TIMP-2 in Leydig cells in old yaks suggested that both may be involved in the interstitial metabolism of the testes during this period. This study revealed the possible role of MMP-2 and TIMP-2 in testicular functionality of yaks at different ages.

The TGFβ system and TIMP1 and 3 genotypes in Turner syndrome-Relation with aortic congenital malformations.

Cardiovascular complications and congenital malformations are known traits in Turner syndrome (TS), which increases mortality. Women with TS have varying phenotype and cardiovascular risks. A biomarker assessing the risk for cardiovascular complications could potentially reduce mortality in high-risk TS and reduce screening in TS participants with low cardiovascular risk. As part of a study initiated in 2002, 87TS participants and 64 controls were invited to magnetic resonance imagingof the aorta, anthropometry, and biochemical markers. TS participants were re-examined thrice lastly in 2016. The focus of this paper is the additional measurements of transforming growth factor beta (TGFβ), matrix metalloproteinase (MMP's), tissue inhibitor of matrix metalloproteinase (TIMP), peripheral blood DNA and their associations with TS and the cardiovascular risk and congenital heart disease. TS participants had lower TGFβ1 and TGFβ2 values compared to controls. snp11547635 heterozygosity was not associated with any biomarkers but was associated with increased risk of aortic regurgitation. TIMP4 and TGFβ1 were correlated with the aortic diameter at several measuring positions. During follow-up, the antihypertensive treatment decreased the descending aortic diameter and increased TGFβ1 and TGFβ2 levels in TS. TGFβ and TIMP's are altered in TS and may play a role in the development of coarctation and dilated aorta. snp11547635 heterozygosity was not found to impact biochemical markers. Further studies should investigate these biomarkers to further unravel the pathogenesis of the increased cardiovascular risk in TS participants.

Effect of Homocysteine on the Expression of Matrix Metalloproteinase-9 and Apoptosis in Trabecular Meshwork Cells

Purpose: In retinal endothelial cells, homocysteine (Hcy) activates matrix metalloproteinase (MMP)-9, which results in apoptosis. This study investigated these effects of Hcy in human trabecular meshwork cells (HTMC).Methods: HTMC cultures using 5 mM low or 20 mM high glucose (HG)-containing media were exposed to 100 μM Hcy for 3 days. Cell viability was assessed with the MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) assay. The MMP-9 and tissue inhibitor of MMP (TIMP)-1 levels were measured by western blotting and the degree of apoptosis was analyzed with flow cytometry using Annexin-propidium iodide double staining.Results: Exposure to Hcy in HG decreased cell viability compared to HG alone (&lt;i&gt;p&lt;/i&gt; = 0.036). Compared to HG alone, co-exposure to Hcy with HG decreased the TIMP-1 levels, albeit not significantly (&lt;i&gt;p&lt;/i&gt; = 0.094), and did not affect the MMP-9 levels (&lt;i&gt;p&lt;/i&gt; = 0.413). In addition, co-exposure to Hcy with HG produced no difference in the degree of apoptosis compared to HG alone (&lt;i&gt;p&lt;/i&gt; = 0.437).Conclusions: Unlike in retinal endothelial cells, Hcy did not affect the activities of TIMP-1, MMP-9, or the degrees of apoptosis significantly in HTMC. Thus, the effects Hcy may be limited in HTMC.

Histopathological evaluation of uterine structure and immunohistochemical examination of MMP-1 and TIMP-1 in cows recovered from metritis

Objective: The aim of this study is to compare the results of histopathological examination of the uterus of healthy cows and the uterus of cows that have recovered from metritis. &#x0D; Materials and Methods: The study materials obtained from the slaughterhouse were divided into two groups: the uterus of healthy cows (no metritis; Group N; n=10) and uterus of cows that had recovered from metritis about 45 days ago (metritis; Group M; n=10). Sections were taken from the uterus of both groups and were stained with hematoxylin and eosin (H&amp;E) for pathological comparison. In addition, the sections were stained immunohistochemically for the examination of Matrix metalloproteinases 1 (MMP-1) and the tissue inhibitor of matrix metalloproteinases (TIMP-1) levels. &#x0D; Results: The uterus showed pathological condition in Group M than in Group N. MMP1 immunopositivity was higher in the luminal epithelium (p

TIMP1 represses sorafenib-triggered ferroptosis in colorectal cancer cells by activating the PI3K/Akt signaling pathway

Background Ferroptosis is involved in the drug resistance mechanisms of some tumors. The present study aimed to explore the role of tissue inhibitor of matrix metalloprotease 1 (TIMP1) in sorafenib-triggered ferroptosis in colorectal cancer (CRC). Methods HCT-8 CRC cell lines were generated that were sorafenib-resistant or that under- or overexpressed TIMP1. The levels of reactive oxygen species (ROS), iron, and malondialdehyde (MDA) were compared across the different cell lines. The half-maximal inhibitory concentration of sorafenib against the different lines was determined based on cell viability. Expression of ferroptosis-related genes and the corresponding proteins was determined by quantitative RT-PCR or western blotting. Results TIMP1 overexpression induced sorafenib resistance in HCT-8 cells. TIMP1 knockdown repressed the activation of the PI3K/Akt pathway and reduced levels of glutathione peroxidase 4 (GPX4), enhancing sorafenib-induced ferroptosis. This led to accumulation of ROS, iron, and MDA. Giving sorafenib and the GPX4 inhibitor RSL3 to sorafenib-resistant HCT-8 cells induced ferroptosis, leading to elevated levels of iron and lipid peroxides, ultimately reducing cell viability. TIMP1 depletion in CRC cells enhances sorafenib-triggered ferroptosis by reducing PI3K/Akt axis signal transduction. Conclusion The combination of sorafenib and GPX4 inhibitors such as RSL3 may be a promising therapy against CRC.

Effect study of exosomes derived from platelet-rich plasma in the treatment of knee cartilage defects in rats

BackgroundThe repair of articular cartilage defects has always been a difficult problem. We aimed to investigate the therapeutic effect of intra-articular injection of platelet-rich plasma (RPR) and PRP-derived exosomes (PRP-Exos) on cartilage defects in rat knee joints and then provide experience for the use of PRP-exos in cartilage defect repair.MethodsRat abdominal aortic blood was collected, and PRP was extracted by two-step centrifugation. PRP-exos were obtained by kit extraction, and PRP-exos were identified by various methods. After the rats were anesthetized, a cartilage defect subchondral bone was created at the proximal end of the origin of the femoral cruciate ligament with a drill. SD rats were divided into 4 groups, including PRP group, 50 μg/ml PRP-exos group, 5 μg/ml PRP-exos group, and control group. One week after the operation, 50 μg/ml PRP, 50 μg/ml PRP-exos, 5 μg/ml PRP-exos and normal saline were injected into the knee joint cavity of rats in each group, once a week. A total of two injections were given. On the 5th and 10th week after drug injection, the serum levels of matrix metalloproteinase 3 (MMP-3) and tissue inhibitor of matrix metalloproteinase 1 (TIMP-1) were detected by each treatment method, respectively. The rats were killed at the 5th and 10th weeks, respectively, and the cartilage defect repair was observed and scored. The defect repair tissue sections were used for HE staining and type II collagen immunohistochemical staining.ResultsThe histological results showed that both PRP-exos and PRP could promote cartilage defect repair and type II collagen formation, and the promoting effect of PRP-exos was significantly better than that of PRP. In addition, enzyme-linked immunosorbent assay (ELISA) results showed that compared with PRP, PRP-exos could significantly increase serum TIMP-1 and decrease serum MMP-3 in rats. And the promoting effect of PRP-exos was concentration dependent.ConclusionIntra-articular injection of PRP-exos and PRP can promote the repair of articular cartilage defects, and the therapeutic effect of PRP-exos is better than the same concentration of PRP. PRP-exos are expected to be an effective treatment for cartilage repair and regeneration.

Interleukin-17 and Tumor Necrosis Factor Show a Functional Hierarchy to Regulate the Production of Matrix Metalloproteases by Monocytes from Patients with Psoriasis.

Interleukin-17 (IL-17) and tumor necrosis factor (TNF) regulate tissue remodeling through matrix metalloproteinases (MMPs). It is not yet clear whether these cytokines have a functional hierarchy in psoriasis. Serum levels of TNF (1,403 versus 1,058 pg/mL), IL-17 (1,528 versus 820 pg/mL), MMP-1 (1,999 versus 1,039 pg/mL), and MMP-9 (1,950 versus 1,561 pg/mL) were higher in psoriasis subjects (n = 60) than in control subjects (n = 60). Tissue inhibitor of MMPs (TIMP-1; 1,374 versus 1,218 pg/mL) was lower in psoriasis subjects. Serum IL-17 was correlated with MMP-2 (rs = 0.40) and TIMP-1 (rs = -0.26) levels. Unstimulated production of MMP-1, MMP-2, and MMP-9 by monocytes was higher in psoriasis subjects, whereas TIMP-1 production was lower. TNF stimulation increased all MMPs, whereas TIMP-1 production was unchanged. IL-17 stimulation increased all MMPs, whereas TIMP-1 production was decreased in psoriasis subjects. MMP-9 production was higher in monocytes stimulated with IL-17 compared with TNF. TIMP-1 production was decreased more by IL-17 than by TNF, but only in psoriasis cells. MMP-1/TIMP-1, MMP-2/TIMP-1, and MMP-9/TIMP-1 ratios were higher after IL-17 stimulation (compared with TNF stimulation) in psoriasis subjects; this occurred in controls only for the MMP-2/TIMP-1 ratio. IL-17 has a greater ability than TNF to dysregulate the MMPs/TIMP-1 balance, supporting IL-17 blockade as first-line treatment in cutaneous psoriasis.

Lung function and collagen 1a levels are associated with changes in 6 min walk test distance during treatment of TB among HIV-infected adults: a prospective cohort study

BackgroundPatients with tuberculosis (TB) and HIV often present with impairments in lung function and exercise capacity after treatment. We evaluated clinical and immunologic variables associated with a minimum clinically important difference (MCID) in the change in the 6 min walk test distance during the first 24 weeks of antiretroviral (ART) and anti-tubercular therapy.MethodsAdults initiating ART and anti-TB treatment in the setting of newly-diagnosed HIV and pulmonary TB were enrolled in a prospective cohort study in South Africa. Patients underwent 6 min walk tests and spirometry at weeks 0, 4, 12, and 24 and biomarker level measurements early during treatment, at weeks 0, 4, and 12, when inflammation levels are typically elevated. Biomarkers included matrix metalloproteinases-1 (MMP-1), tissue inhibitor of MMP (TIMP)-1, collagen 1a, IL-6, IL-8, vascular cell adhesion molecule 1 (VCAM-1), C-X-C motif chemokine 10 (CXCL-10), CXCL-11, macrophage colony-stimulating factor (M-CSF), plasminogen activator, vascular endothelial growth factor, and chemokine (C-C) motif-2 (CCL-2). An MCID was derived statistically, and achievement of an MCID was modeled as the outcome using logistic regression model.ResultsEighty-nine patients walked an average of 393 (± standard deviation = 69) meters at baseline, which increased by an average of 9% (430 ± 70 m) at week 24. The MCID for change in walk distance was estimated as 41 m. Patients experiencing an MCID on treatment had worse lung function, lower 6 min walk test distance, higher levels of proinflammatory biomarkers including TIMP-1 and M-CSF, and lower levels of collagen 1a at baseline. Experiencing an MCID during treatment was associated with increases in forced expiratory volume in 1-s [odds ratio (OR) = 1.17, 95% confidence interval (CI) = 1.05–1.33] and increases in blood collagen 1a levels (OR = 1.31, 95%CI 1.06–1.62).ConclusionsART and TB treatment are associated with substantial improvements in 6 min walk test distance over time. Achievement of an MCID in the 6 min walk test in this study was associated with more severe disease at baseline and increases in collagen 1a levels and lung function during therapy.

TIMP-1 as a biomarker in obstructive sleep apnea: screening, monitoring, risk stratification, and a step towards precision medicine.

The diagnosis of obstructive sleep apnea (OSA) is a complex time- and resource-intensive diagnostic procedure. Since tissue inhibitors of matrix metalloproteinases (TIMP's) are involved in various pathophysiological processes and are correlated with a high cardiovascular risk, TIMP's appear to be a suitable candidate for an OSA-biomarker. In a prospective controlled diagnostic study, TIMP-1 serum levels of 273 OSA-patients and controls were analyzed for correlation with OSA severity, BMI, age, sex, cardio-/ cerebrovascular comorbidities. Furthermore, longitudinal medium- and long-term effects of CPAP-treatment (n=15) on TIMP-1-levels were investigated. TIMP-1 was clearly linked to OSA as well as to disease severity (mild, moderate, severe; each p<0.001) and was not influenced by age, gender, BMI, or cardio-/cerebrovascular comorbidities. ROC curve analysis revealed an AUC of 0.91 ± 0.017 SE (p<0.001), suggesting a TIMP-1 cut-off value of 75 ng/ml (sensitivity 0.78; specificity 0.91) being especially sensitive for patients with severe OSA (sensitivity 0.89; specificity 0.91). The likelihood ratio was 8.88, while the diagnostic odds ratio was 37.14. CPAP-treatment led to a significant decrease of TIMP-1 after 6-8 months (p=0.008). TIMP-1 seems to fulfill the preconditions for a circulating OSA-biomarker: disease-specific with a mandatory presence in affected patients, reversible on treatment, reflects disease severity and provides a cutoff value between the healthy state and disease. In the clinical routine, TIMP 1 may help to stratify the individual OSA-associated cardiovascular risk and to monitor the treatment response to CPAP-therapy as a further step towards providing a personalized therapy.

СРАВНИТЕЛЬНЫЙ АНАЛИЗ КЛИНИКО-АНАМНЕСТИЧЕСКИХ И ЛАБОРАТОРНО-ИНСТРУМЕНТАЛЬНЫХ ХАРАКТЕРИСТИК ПАЦИЕНТОВ С ИШЕМИЧЕСКОЙ БОЛЕЗНЬЮ СЕРДЦА, ПРОЖИВАЮЩИХ В УСЛОВИЯХ КРАЙНЕГО СЕВЕРА И УМЕРЕННОЙ КЛИМАТИЧЕСКОЙ ЗОНЫ

Introduction.Climatic conditions of the Far North are a factor in the early development and progression of coronary artery disease. Aim. To study the clinical-anamnestic and laboratory-instrumental characteristics of patients with coronary artery disease living in the Far North and the temperate climate zone. Material and methods. An observational analytical study of 302 patients with coronary artery disease admitted to the hospital for coronary angiography was performed. Group 1 (n=168) included patients living in the Yamalo-Nenets Autonomous Okrug, group 2 (n=134) included patients living in the temperate climate zone. The clinical status of patients, drug therapy, laboratory data, and coronary angiography results were assessed. Student’s t-test, Mann-Whitney U-test, χ2 test or Fisher’s exact test were used when comparing indicators between the groups.. Results and discussion. In group 1, despite the younger age of patients, a history of myocardial infarction and chronic heart failure with a low left ventricular ejection fraction were more often recorded, which may be due to the so-called “polar tension syndrome”, which triggers pathological mechanisms of remodeling of the cardiovascular system. In elderly patients in group 2, hypertensive crises were more often recorded, and systolic blood pressure was higher and did not reach the target values. There were no differences in the nature of the lesion of the coronary bed. In both groups, a high incidence of hyperlipidemia was established and elevated levels of homocysteine, matrix metalloproteinase-9 and tissue inhibitor of matrix metalloproteinase-1, which are markers of atherosclerotic plaque instability and rapid progression of atherosclerosis, were registered. There were no differences in adherence to statin therapy, but the average dose of atorvastatin in group 2 was 2 times lower than in group 1. Group 1 patients were more adherent to taking antiplatelet agents, mainly due to middle-aged patients. Conclusion. In patients with coronary artery disease living in the Far North, despite their younger age, myocardial infarction and severe heart failure were more often found. All patients had a high incidence of hyperlipidemia and elevated levels of immune inflammation biomarkers. Regional differences in prescribed therapy and in the formation of patients’ adherence to treatment were revealed.

The Relationship Between Thiamin Administration And Lactic Acid In Sepsis Patients In The Icu Of H. Adam Malik Hospital

Background: Thiamine may be considered as adjunctive therapy in patients with sepsisThiamine administration reduced the Matrix metalloproteinase 9 / Tissue inhibitors of matrix metalloproteinase 1 (TIMP-1/MMP-9) ratio, which could be associated with better survival.

Doxycycline-Induced Changes in Circulating MMP or TIMP2 Levels Are Not Associated with Skeletal-Related Event-Free or Overall Survival in Patients with Bone Metastases from Breast Cancer.

Doxycycline is often used as a promoter of inducible gene expression in preclinical models; however, it can also have direct effects on tumor growth and survival. This is due in part to its ability to inhibit cell invasion and regulate matrix metalloproteinase (MMP) expression. Given that doxycycline is also osteotropic, a clinical study to assess its effects on modulation of tumor progression or prevention of skeletal-related events (SRE) in patients with bone metastases from breast cancer (the Achilles trial) was undertaken. Patients received 100 mg of oral doxycycline twice daily for 12 weeks, with serum obtained at baseline and 4, 8 and 12 weeks post-initiation of doxycycline treatment. Exploratory analysis of the effects of doxycycline on circulating levels of MMP or tissue inhibitor of matrix metalloproteinase 2 (TIMP2) was performed in enrolled patients. Statistically significant associations were observed between MMP2, MMP9 and TIMP2 at baseline with significant associations maintained between absolute levels and changes in levels of MMP2 and TIMP2 at weeks 4-12 post initiation of doxycycline. Treatment with doxycycline generally resulted in decreases in MMP2 and MMP9 levels with concurrent upregulation of TIMP2 at 12 weeks post-initiation of doxycycline treatment. Despite this, we observed no association with the levels of any of these factors with either SRE-free or overall survival in this patient cohort. In summary, despite observing hypothesized effects of doxycycline administration on surrogate markers of its anti-tumor activity, measures of circulating levels of these biomarkers were not prognostic in this patient population.