Content Of TIMP-1 Research Articles

Liposome-encapsulated dextrazide modifies spleen extracellular matrix composition in mice with chronic BCG-induced inflammation

Fibrosis of parenchymal organs is a common complication of tuberculosis. In a model of BCG-induced inflammation in mice, changes in the metabolism of the extracellular matrix (ECM) of the spleen were demonstrated with the introduction of a liposome-encapsulated dextrazide (LEDZ) containing isoniazid and oxidized dextran.The mice were divided into 4 groups: 1 – intact animals; 2 – infected mice after a single intravenous injection of BCG vaccine. 6 mo after infection, a solution of LEDZ was administered intraperitoneally to mice of group 3 for 3 mo, and inhaled to mice of group 4. Group 2 mice showed the signs of pronounced spleen fibrosis (increased content of hyaluronan, hydroxyproline fractions) with activation of hyaluronidases, matrix metalloproteinases (MMP), α2-macroglobulin and an increased content of tissue inhibitors of MMP (TIMP-1 and TIMP-2) with respect to group 1 data. In group 3, changesin the structure of proteoglycans were noted (an increase in the content of uronic acids and galactose), a decrease in the content of hyaluronan and free hydroxyproline, an increase in the activity of hyaluronidases. The MMP activity and the TIMP content corresponded to the data of group 2. In group 4, the content of uronic acids and galactose in proteoglycans also increased, but peptide-bound hydroxyproline decreased and the hyaluronan content more noticeably decreased. The activity of all enzymes regulating the ECM metabolism reduced with respect to the data of group 2.Thus, intraperitoneal administration of LEDZ to infected mice led to activating hyaluronidases, changing the structure of proteoglycans, and decreasing the free hydroxyproline content. Inhalation administration of LEDZ, along with changes in the structure of proteoglycans, reduced the activity of MMP, hyaluronidases, α2-macroglobulin, the content of TIMP-1 and TIMP-2, peptide-bound hydroxyproline. The antifibrotic effect of LEDZ with inhalation administration was manifested in a decrease in peptide-bound hydroxyproline and in a more significant decrease in hyaluronan compared with intraperitoneal administration.Thus, intraperitoneal administration of LEDZ to infected mice led to activating hyaluronidases, changing the structure of proteoglycans, and decreasing the free hydroxyproline content. Inhalation administration of LEDZ, along with changes in the structure of proteoglycans, reduced the activity of MMP, hyaluronidases, α2-macroglobulin, the content of TIMP-1 and TIMP-2, peptide-bound hydroxyproline. The antifibrotic effect of LEDZ with inhalation administration was manifested in a decrease in peptide-bound hydroxyproline and in a more significant decrease in hyaluronan compared with intraperitoneal administration.

Evaluation of the influence of Inosine pranobex on the matrix protein system in patients with chronic viral cervicitis

The reproductive potential of both women and men is declining every year. Many factors contribute to the violation of the reproductive function – chemical, physical, mechanical, psychogenic, however, biological factors have the most pronounced effect on reproduction. Chronic viral cervicitis can be not only the cause of infertility and reproductive losses, but also the development of intraepithelial dysplasia, as well as cervical cancer. PVI, as a monoinfection, is quite rare along with HPV. Other UGIs (urogenital infections) act as common routes of transmission and entry gates. The most common association with PVI is herpesvirus infection. An increase in MMP, both systemically and at the local level, may indicate a violation of cell modeling processes, which contributes to the development of autoimmune inflammation with further destruction of the tissues of the reproductive tract. Activation of MMP promotes the release of HSV from the nerve ganglia and reactivation of the infection. Therapy for HPV and HVI (herpes virus infections) are debatable. There is no single standard of treatment, but there are a number of drugs that have antiviral and immunomodulatory effects. Currently, there are no studies on the dynamics of the effect of HPV and HSV infection on the state of MMPs and TIMPs during Inosine pranobex therapy. Objective: to evaluate changes in matrix metalloproteinases 2 and 9 and their tissue inhibitors types 1 and 2 in patients with human papillomavirus and herpes infections after Inosine pranobex therapy. 6 patients with papillomavirus and herpetic infections were examined and treated with drugs containing the active ingredient Inosine pranobex. The levels of MMP-2, MMP-9 and TIMP-1, TIMP-2 in blood serum were determined using specific reagents from R&D Diagnostics Inc. (USA). The dynamics of indicators in the blood serum of patients with PVI showed a decrease in the level of MMP-2, MMP-9, TIMP-1 with a simultaneous increase in TIMP-2 relative to the values before therapy. In patients with PVI and HVI, Inosine pranobex therapy showed a decrease in MMP-2 and MMP-9 levels, no changes in the content of TIMP-1, but an increase in the serum content of TIMP-2. Prior to the use of therapy, an increase in the ratio in the main groups in comparison with the control group was found, however, the largest increase was found in the group with the association of infections. After therapy, positive dynamics was established in the main groups. Thus, the ratio in group I decreased and became equal to the control values. In the II group of patients, the ratio, despite the decrease, remained higher than the control values and higher in comparison with the I group of women.

Abstract 2356: Aberrant TIMP-1 secretion by tumor-associated fibroblasts is a major contributor to the selective positive response to nintedanib in lung adenocarcinoma

Abstract Adenocarcinoma (ADC) and Squamous Cell Carcinoma (SCC) are the most frequent histologic subtypes of lung cancer, and both are rich in activated/myofibroblast-like tumor-associated fibroblasts (TAFs). Nintedanib is a potent antifibrotic drug that targets the tumor stroma and is clinically approved to treat advanced lung ADC patients owing to the survival benefits observed in the LUME-1 clinical trial in ADC but not SCC patients. Although the mechanism underlying the ADC-selective therapeutic effects of nintedanib remained poorly understood, we have previously reported that nintedanib abrogates the pro-tumoral traits of the secretome of ADC-TAFs but not SCC-TAFs, suggesting that secreted factor(s) in ADC-TAFs may be implicated. In addition, we recently unveiled an ADC-specific tumor-promoting crosstalk between TAFs and cancer cells driven by TIMP-1 and CD63, whereas others have shown that TIMP-1 is a molecular target of nintedanib. However, it remains unknown if TIMP-1 is involved in the ADC-selective benefits of nintedanib. To address this question, we used patient-derived TAFs obtained with the patient’s informed consent and using protocols approved by the Ethics Committee of the Hospital Clinic. We treated TGF-β1-activated ADC-TAFs and SCC-TAFs with nintedanib and determined the content of TIMP-1 in their conditioned medium by ELISA. Moreover, we used cell-based functional assays and tumor xenografts after knocking-down TIMP-1 in TAFs by siRNA to examine how silencing TIMP-1 altered their response to nintedanib. By analyzing TCGA data and the human protein atlas, we found that TIMP-1 is consistently upregulated in ADC compared to SCC tumors both at the mRNA and protein levels. Moreover, TCGA data revealed that TIMP-1 is increased in tumors compared to paired control tissue in ADC but not SCC. In culture, we observed that TIMP-1 secretion was higher in ADC-TAFs than SCC-TAFs, and that this secretion was downregulated by nintedanib to a larger extent in ADC-TAFs compared to SCC-TAFs. In vivo analyses revealed that ADC cells co-injected with fibroblasts silenced for TIMP-1 as in SCC-TAFs into immunocompromised mice exhibited a less invasive growth pattern compared to tumors bearing control fibroblasts. In addition, we observed that the inhibition of the pro-tumoral secretome of ADC-TAFs elicited by nintedanib was abrogated upon knocking-down TIMP-1 in TAFs both in vitro and in vivo. Collectively, these results reveal that the high secretion of TIMP-1 in ADC-TAFs render them more responsive to nintedanib, whereas the low TIMP-1 secretion of SCC-TAFs may be a major contributor to the selective resistance of SCC patients to nintedanib. Moreover, our results identify TIMP-1 as a promising predictive biomarker of nintedanib responses. Citation Format: Paula Duch, Natalia Díaz-Valdivia, Marta Gabasa, Rafael Ikemori, Frank Hilberg, Noemí Reguart, Derek Radisky, Jordi Alcaraz. Aberrant TIMP-1 secretion by tumor-associated fibroblasts is a major contributor to the selective positive response to nintedanib in lung adenocarcinoma [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 2356.

Dynamics of matrix metalloproteinases when correcting immature hypertrophic scars with pulsed dye laser and fermencol phonophoresis

The article analyzes the role of matrix metalloproteinases (MMP) in the pathogenesis of immature hypertrophic scars. The revealed decrease in the MMP level in the patients’ initial state against the background of an increased content of TIMP1 indicates a disorder in the mechanisms of regulation of collagen formation in a hypertrophic scar, as a result of which its synthesis significantly predominates over decay, contributing to the formation of a fibrous process. The obtained results convincingly prove that reduced expression of MMP against the background of an increased TIMP1 level is an important pathogenetic mechanism providing excessive deposition of extracellular matrix components in pathological skin scarring. The use of a pulsed dye laser (PDL) in combination with Fermencol phonophoresis promotes an increase in the MMP content in the blood serum of patients with immature hypertrophic scars. The reliable dynamics of the studied biomarkers indicates an increase in the processes of catabolism of the extracellular matrix components and an increase in the regenerative potential of the skin defect, which determine the intensity of the clinical effect during the treatment of immature hypertrophic scars. The correlation analysis between the level of membrane metalloproteinases and the parameters of the Vancouver Scar Scale revealed a set of significant relationships that confirm the clinical and pathogenetic significance of these biochemical variables in the development of fibroplastic processes and also act as informative criteria for the effectiveness of the therapy.

Content of matrix metalloproteinases in the blood of hypertensive patients with a high cardiovascular risk receiving statin therapy

Aim. To compare the effect of atorvastatin and rosuvastatin as part of complex therapy in patients with hypertension (HTN) with a high cardiovascular risk on the level of matrix metalloproteinases -1, -9 (MMP-1, MMP-9) and tissue inhibitors of metalloproteinases -1, -4 (TIMP-1, TIMP-4).Material and methods. The study included 140 hypertensive patients who received atorvastatin (Liprimar) 20 mg/day in addition to antihypertensive therapy for a year, which was later replaced by rosuvastatin (Rosucard) in the following doses: 10 mg/day (n=96), 20 mg/day (n=24), 40 mg/day (n=26). Patients underwent standard clinical and paraclinical investigations. In the blood serum of patients, the levels of MMP-1, MMP-9 and TIMP-1, TIMP-4 were determined.Results. Patients who used rosuvastatin at a dose of 40 mg/day had a more pronounced decrease in MMP-1 than those treated with rosuvastatin at a dose of 10 and 20 mg/day (p<0,05), while there were no differences in MMP-1 when using low and medium doses. Rosuvastatin had a less pronounced effect on MMP-9 than on MMP-1, while increasing the dose of rosuvastatin did not affect the intensity of MMP-9 reduction (p>0,05). The content of TIMP-1 and TIMP-4 increased when taking rosuvastatin, while a more pronounced dose-dependent increase in TIMP-1 was observed with rosuvastatin 20 mg/day and 40 mg/day. In addition, the largest increase in TIMP-4 was observed when using rosuvastatin at a dose of 40 mg/day. Atorvastatin had no significant effect on MMP-1 and MMP-9, as well as TIMP-1 and TIMP-4.Conclusion. Long-term rosuvastatin therapy (10 mg/day, 20 mg/day, 40 mg/day) as part of the complex therapy of cardiovascular patients affects the metabolism of vascular wall elastin and collagen, reducing the level of MMP-1, MMP-9 and increasing the content of TIMP-1, TIMP-4 in the blood.

Specific Parameters of Extracellular Matrix Remodeling of Liver and Lungs of Mice with BCG Granulomatosis during Chronic Inflammation Depending on the Method of Administration of Liposomal Oxidized Dextran

The objective: to study the effect of liposomal oxidized dextran (LOD) on remodeling of the extracellular matrix of organs of mice during the period of chronic BCG granulomatosis and effects of LOD on this process depending on the route of administration. Subjects and Methods. The liver and lungs of mice with BCG granulomatosis were studied using different methods of LOD administration (intraperitoneally and by inhalation). The content of glycosaminoglycans, hydroxyproline fraction, the activity of matrix metalloproteinases (MMPs), hyaluronidases and a2-macroglobulin, and the content of tissue inhibitors of MMPs (TIMP-1 and -2) were evaluated. Results. The administration of LOD to mice infected with the BCG vaccine suffering from chronic granulomatosis with severe fibrosis (6 months after infection for 3 months) resulted in the aggravation of collagen degradation in the liver. In the lungs, along with increased collagen degradation, decreased collagen synthesis was observed. It was apparently due to suppressed activity of a2-macroglobulin and decreased content of TIMP-1 and -2. In the liver, with intraperitoneal administration of LOD, signs of suppressed fibrogenesis and fibrolysis were observed versus the data obtained for inhalation administration. There were no differences in the content of hydroxyproline fractions in the lungs depending on the method of LOD administration. Thus, administration of LOD to mice led to lower severity of fibrosis while the mechanisms of fibrolysis in the lungs and liver differed.

Matrix metalloproteinases in children with uncomplicated compression fractures of the spine

The aim of the study was to determine changes in the content of matrix metalloproteinases (MMP) and their tissue inhibitor in children with uncomplicated compression fractures of the spine (UCFS). Materials and methods. Eighty-five children, including 69 patients with UCFS (average age 12.3 ± 2.6 years), were comprehensively examined. The reference group consisted of 16 children (average age 11.8 ± 2.7 years) without spinal pathology. During the diagnostic period for 1-3 days, changes in the MMP content and their tissue inhibitor (TIMP-1) in blood serum were determined by the enzyme immunoassay method in all children after trauma. Results. It was found that in the acute period after spinal injury, the blood levels of gelatinases (MMP-2 and MMP-9), stromelysin (MMP-3), and collagenases (MMP-8) significantly increased compared to their levels in children of the reference group. At the same time, the levels of TIMP-1 and the ratio of MMP/TIMP-1 concentrations in the blood of patients with UCFS significantly decreased compared to the control, which indicates the predominance of the proteolytic effect of MMP. Analysis of changes in the content of MMP in the blood in UCFS boys and girls did not reveal significant differences in the levels of the studied MMP and TIMP-1, except for a significant increase in the concentrations of stromelysin (MMP-3) in the blood serum of boys compared with its level in girls and the control. With different severity of the course of UCFS in children, a significant increase in MMP concentrations associated with an increase in the severity of the injury was revealed, and a substantial decrease in the content of TIMP-1 in the blood of patients compared to its levels in children with 1-2 degrees of severity and control. Conclusion. The established patterns indicate that the determination of the content of MMR and TIMP-1 in the blood in UCFS children allows monitoring the course of the reparative process after injury to the vertebral bodies in children.

Effect of Gancao Fuzi Decoction on osteoarthritis and proteomics of articular cartilage in rats

The rat osteoarthritis model was replicated by injection of sodium iodoacetate into the knee joint cavity, and the effects of Gancao Fuzi Decoction on rat osteoarthritis and the proteome of articular cartilage were investigated. Sixty SD rats weighing 230-250 g were randomly divided into normal group, model group, glucosamine sulfate group, and Gancao Fuzi Decoction high, medium and low dose groups. Osteoarthritis model was induced by intra-articular injection of sodium iodoacetate(3 mg on each leg) in all groups except the normal group. After modeling, each administration group was given intragastric administration for 1 month. During the administration period, joint pain test and joint width measurement were performed every week to observe the autonomous behavior of rats. Enzyme linked immunosorbent assay(ELISA) method was used to detect the contents of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), matrix metalloproteinase-3(MMP-3), and matrix metalloproteinase tissue inhibitor(TIMP-1) in rat joint lavage fluid. Hematoxylin-eosin(HE) staining was used to observe bone and joint morphology. Nano-LC-LTQ-Orbitrap system was used to detect arti-cular cartilage proteins. The results showed that, compared with the model group, Gancao Fuzi Decoction could significantly improve joint pain and joint swelling in osteoarthritis rats, significantly reduce the contents of TNF-α, IL-1β and MMP-3 in the joint cavity la-vage fluid, increase the content of TIMP-1, and relieve inflammatory diseases such as enlarged joint space, rough cartilage edge, different thickness of cartilage layer, and disordered arrangement of chondrocytes. After comparing the proteins between the groups, 273 differential proteins were screened out. KEGG analysis found that the above differential proteins involved 43 signaling pathways such as systemic lupus erythematosus, among which 11 signaling pathways were related to osteoarthritis. The above results indicated that Gancao Fuzi Decoction had a preventive effect on osteoarthritis, and its mechanism of action may be accomplished by regulating the protein expression of osteoarthritis-related signal pathways.

Activity of matrix metalloproteinases and their tissue inhibitors in serum in chronic granulating periodontitis

Aim. The aim of the work is to study the activity of matrix metalloproteinases and their tissue inhibitors in chronic granular periodontitis. Materials and methods. We examined serum of 15 patients with chronic granulating periodontitis and 15 healthy male blood donors aged 32 to 37 years. Determination of MMP-8, MMP-9 and TIMP-1 activity was performed by solid-phase enzyme immunoassay using test systems (R&D Diagnostics Inc., USA) on the automatic immunoenzyme complex “GBG Star Fax 2100” produced by “Awareness Technology Inc.” (USA). Statistical evaluation of the results was carried out using the generated database in the program Statistica v. 10.0 (StatSoft. Inc., USA), № STA999K347156-W. Results. It was established that in the phase of exacerbation of chronic granulating periodontitis in the blood serum of patients there was an increase in concentrations of MMP-8 and MMP-9, which exceeded the content of TIMP-1, which was accompanied by a decrease in the ratio of MMP-8/TIMP-1 and MMP-9/TIMP-1, compared with those for healthy individuals. In the phase of early convalescence, 3–5 days after treatment of patients with exacerbation of chronic granulating periodontitis, concentrations of MMP-8 and MMP-9 in the blood serum of the examined patients decreased more dynamically than the concentration of TIMP-1, that was accompanied by an increase in the ratio of MMP-8/TIMP-1 and MMP-9/TIMP-1, at the same time the full normalization of MMP-8, MMP-9, TIMP-1 and the ratio of MMP-8/TIMP-1 and MMP-9/TIMP-1 did not happen. Conclusions. Chronic granulating periodontitis is accompanied by changes in the blood serum of patients with MMP-8, MMP-9 and TIMP-1, which depend on the phase of the pathological process. The highest concentrations of MMP-8, MMP-9 and TIMP-1 are recorded in the acute phase of the disease. In the phase of early convalescence, 3–5 days after the remediation of the focus of inflammation, the concentrations of MMP-8, MMP-9 and TIMP-1 in serum are decreased, while the dynamics of decreasing of the content of MMP-8 and MMP-9 prevails over that for TIMP-1. The coefficients MMP-8/TIMP-1 and MMP-9/TIMP-1, which characterize the balance in the system of MMP/TIMP, allow assessing the state of the system in the dynamics of the disease.

Effect of Liposomal Delivery of Oxidized Dextran with Isonicotinic Acid Hydrazide on Component Profile of Pulmonary Extracellular Matrix in Mice with BCG-Induced Granulomatosis.

Intraperitoneal injections of isonicotinic acid hydrazide (INH), dextrazide (oxidized dextran+INH), or liposomes loaded with dextrazide (INH dose of 14 mg/kg) over 2 months to mice with BCG-induced granulomatosis started from postinfection day 90 induced qualitative and quantitative changes in composition of pulmonary extracellular matrix. Both dextrazide and its liposomal form decreased the levels of sulfated glycosaminoglycans and uronic acids. In contrast to INH, both preparations did not decrease the levels of total glycosaminoglycans, proteins, and galactose. This difference is explained by the fact both free and liposomal dextrazide activated MMP, but did not increase the content of TIMP-1 and TIMP-2, whereas injection of INH was followed by an increase in TIMP-2 content and a decrease in the level of free hydroxyproline, which attested to down-regulation of collagen degradation and maintenance of the conditions for pulmonary fibrosis in mice of this group.

MATRIX METALLOPROTEINASES IN CHILDREN WITH A COMBINED BONE INJURY

There were comprehensively examined 55 children, including 35 children with combined bone injury (average age of 12.6 ± 2.3 years), reference group consisted of 20 apparently healthy children (average age of 11.8 ± 2.7 years) without pathology of the motor system. on the 1-3rd and 30th day after the combined bone injury, changes in the content of matrix metalloproteinases (MMP) and cytokines - transforming growth factor-beta (TGF-β), monocytic chemotactic factor (MCP-1) and macrophage inflammatory protein (MIP-1β) were followed in the dynamics by serum immunoassay determination. It was established that after combined bone injury at the stage of post-traumatic formation of the regenerate by 30th days the serum concentrations of gelatinases (MMP-2, MMP-9) and collagenases (MMP-8) significantly increased, the levels of stromelysins (MMP-3) did not change, and the content of TIMP-1 decreased. Early detection of changes in the blood content of bone biomarkers during the recovery process after a combined trauma in children allows the timely implementing the correction of disturbances and the choice of the optimal individual treatment tactics for a particular patient, taking into account the features of his bone metabolism.

CHANGES IN THE BLOOD SERUM CONTENT OF BONE BIOMARKERS AND CYTOKINES IN CHILDREN WITH COMBINED TRAUMA

Twenty-nine children (mean age of 12.6 ± 2.3 years) with combined bone trauma were examined. The reference group consisted of 20 conditionally healthy children (mean age of 11.8 ± 2.7 years) without the pathology of the locomotor system. The content of bone biomarkers - osteoprotegerin (OPG), bone isoenzyme of alkaline phosphatase (AP), osteocalcin (OC), hyaluronic acid (HA), as well as matrix metalloproteinases (MMPs) and cytokines - TGF-β, MCP-1 and MIP-1β in serum was determined by the enzyme immunoassay in dynamics: on the 1-3rd, 7-th, 14-th and 30-th days after the trauma. Remodeling of bone tissue after a combined trauma at the stage of formation of the regenerate was established to be characterized by diverse changes in the serum content of bone biomarkers, which are not substantially dependent on the severity of the trauma. At the same time, a significant increase in the concentrations of OPG, AP and HA was combined with a pronounced decrease in the content of OC. At 7-14th days after the injury OC levels were lower by more than 3 times compared with the control, indicating a slowdown in the mineralization of the osteoid and a disturbance in the formation of bone tissue during this period. By 30 days after trauma serum concentrations of gelatinases (MMP-2, MMP-9) and collagenases (MMP-8) increased significantly, stromelysin levels (MMP-3) did not change. By 30th day after the injury serum concentrations of gelatinases (MMP-2, MMP-9) and collagenases (MMP-8) increased significantly, stromelysin levels (MMP-3) did not change, and the TIMP-1 content declined. Early detection of changes in blood levels of bone biomarkers during the process of the recovery after combined trauma in children makes it possible to ensure timely correction of disturbances and choice of optimal individual treatment tactics for the management of a particular patient, taking into account the peculiarities of his bone metabolism

Age-Related Dynamics of the Content of Matrix Metalloproteinases (MMP-1, -2, -3, and -9) and Tissue Inhibitors of Matrix Metalloproteinases (TIMP-1, -2, and -4) in Blood Plasma of Residents of the European Part of Russia’s Arctic Zone

This work presents the results of the first ever study of the enzymes involved in the regulation of extracellular matrix metabolism in the European part of Russia’s Arctic zone. The content of matrix metalloproteinases (MMP-1, -2, -3, and -9) and tissue inhibitors of matrix metalloproteinases (TIMP-1, -2, and -4) in blood plasma was evaluated in 91 men, residents of the European part of the Russian Arctic (67° N) and 14 men, residents of Western Siberia (55° N). The content of MMP-1 and MMP-9, as well as TIMP-1 and TIMP-4 in the blood plasma was higher in northerners relative to the Western Siberia residents. The age-related dynamics of MMPs and TIMPs in northerners had a multidirectional trend. We observed the maximum content of MMP-1 and MMP-9 in the group of 30–39 year olds; MMP-3, in the group of 40–49 year olds. The content of these enzymes tended to decrease with age. We found the maximum content of TIMP-1 and TIMP-2 in the group of 50–59 year olds; TIMP-4, in the group of 60–69 year olds; whereas we observed the minimum content of these enzymes in the group of young men under 29 years old. The hyaluronidase activity in plasma was minimal in the group of 30–39 year olds; it increased with age, reaching a maximum value in the group of 50–59 year olds. We can assume that the age-related imbalance in the MMP/TIMP system (a decrease in MMP content with age and vice versa, an increase in TIMP content in older age groups) is one of the reasons for the age-related increase in interstitial fibrosis and premature aging of northerners.

Abstract 105: Association of Viscoelastic Material Properties and Extracellular Matrix Remodeling in Human Abdominal Aortic Aneurysmal Tissue

Objectives: Predicting rupture of abdominal aortic aneurysm (AAA) requires knowledge of both the rate of extracellular matrix (ECM) degradation and the pulsatile stress on the aortic tissue. The activity of matrix metallopeptidase 9 (MMP9) and its inhibitor, TIMP1, are associated with alterations in aortic ECM but it is unknown if these changes effect the dynamic viscoelastic properties. We hypothesize that increased levels of MMP9 within AAA tissue will be associated with a greater dynamic modulus (E*), as a surrogate of increased aortic wall stress. Methods: Human aneurysmal aortic tissue was obtained at the time of open AAA repair (n=11) and age-matched non-aneurysmal cadavers (n=10). Uniaxial viscoelastic material properties were measured in the circumferential orientation under physiologic preload (110 mmHg) and cyclic strain (± 5%@1Hz). Quantitative histologic and immunohistochemistry were preformed using Fiji imaging software. Aortic MMP9 and TIMP1 content and activity were quantified using western blot and zymography. Results: E* was greater (1862±464 vs 1362±405 kPa, p=0.02) in the AAA tissue as compared to non-aneurysmal tissue. AAA tissue contained less elastin (6.7±6.7 vs 23.4±8.7%, p=0.01) and a greater collagen/elastin ratio (19.9±20.6 vs 2.3±2.5%, p=0.05). Immunohistochemistry revealed 200% greater MMP9 content in the AAA tissue (Figure A & B, 0.61 vs 0.03%, p=0.03). Increased MMP9 content was confirmed using a western blot (0.43 vs 0.06 AU, p<0.01). No difference in relative MMP9 activity (4307 vs 2324 AU, p=0.25) or level of TIMP1 (0.03 vs 0.02, p=0.6) were observed. There was a positive linear correlation (Figure C, r 2 =0.47) between E* and MMP9 as determined by quantitative immunohistochemistry. Conclusions: Our data suggests a positive relationship between E* and MMP9 content. Increased tissue stiffness may trigger MMP9 production resulting in a positive-feedback loop, progressively increasing aortic wall stress and rupture risk.

APPLICATION IMMUNOHISTOCHEMICALLY RESEARCH METHODS IN THE DIAGNOSIS OF PROSTATE CANCER

Introduction. The actual problem of modern urology remains differential diagnostics of various diseases of the prostate gland. The purpose of the study. Increasing the effectiveness of differential diagnosis of diseases of the prostate. Materials and methods. In the biopsy specimens of patients with benign prostatic hyperplasia (BPH) and prostate cancer (PCa), an immunohistochemical study of the production of the Ki-67 proliferation marker, matrix metalloproteinase-9, the matrix metalloproteinase inhibitor TIMP-1, and the distribution of collagen type IV was performed. Results. A moderate positive correlation was found between Gleason gradation and the cell proliferation index for Ki 67 (rs = 0.674) and a moderate negative correlation of Gleason gradation with the level of production of matrix metalloproteinase-9 (rs = -0.660). A weak significant negative correlation was established between the level of proliferative cell activity and the production of MMP-9 tumor cells (rs = -0.369). A significant decrease in the level of MMP-9 and TIMP-1 in adenocarcinoma of different grades was revealed. The invasive properties of tumor cells, expressed in the destruction of collagen of the IV type of the basal membrane and connective tissue prostatic stroma, are mediated by the imbalance between MMP-9 and the protein blocking this enzyme - TIMP-1, whose production decreases in adenocarcinomas of different grades compared with BPH. Conclusions: 1. BPH is characterized by high production of MMP-9 type, which destroys the collagen of the basal membranes and stroma, the proteolytic action of which is blocked by the high content of TIMP-1.

Age-Related Changes in the System Metalloproteinases/Tissue Metalloproteinase Inhibitors and Proteoglycan Components in Mouse Organs.

Activity of MMP and content of TIMP and some proteoglycan components were measured in the liver, lungs, and spleen of BALB/c mice aging 2, 6, and 12 months. The increase in the content of proteoglycan components in mouse organs was associated with age-related changes in MMP activity and TIMP-1 and TIMP-2 levels. The ratio between MMP activity and content of TIMP-1 and TIMP-2 differed in the studied organs. The levels of TIMP-1 and TIMP-2 in mouse blood serum and their concentration in studied organs changed asynchronously with age. They are important regulators that determine MMP activity in the analyzed age periods in mice, which can be determined as periods of growth, development, and ageing.

MARKERS OF CONNECTIVE TISSUE REMODELING IN GENITAL PROLAPSE

Purpose: to expand the conception of molecular and biochemical changes in genital prolapse (GP) based on the study of morphological and immunohistochemical features in connective tissue structures of the ligamentous apparatus of the pelvic floor and their dependence on genetic polymorphisms MMP / TIMP. Materials and Methods: the study involved 178 women aged 35 to 65, 134 of them with GP relapses (after hysterectomy by vaginal access because of a total and partial uterus and vaginal walls prolapse). Patients were randomized into the following groups: I – with manifestations of undifferentiated connective tissue dysplasia (CTD) (11.7 points on average) (n = 86); II – with no CTD signs (n = 48). Control group lll consisted of healthy women without any GP signs (among 15 patients abdominal hysterectomy was performed in connection with uterus hyperplastic processes) (n = 44). Used: morphological method of studies, immunohistochemical (to assess tissue biopsies of sacrum-uterine and round uterine ligaments), the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs), genotyping by polymerase chain reaction of MMP / TIMP polymorphisms. Results: the morphological study of women’s ligamentous apparatus in cases with GP revealed significant fibrosis, coarser collagen septa among bundles of smooth muscle fibers and degenerative changes in individual smooth muscle cells. The group with GP and CTD features showed diffuse atrophy, hyaline or mucinous degeneration of smooth muscle tissue and evident edema of extracellular matrix in 65% of samples. Pathobiochemical disorders in cases of pelvic descencia were determined by an imbalance in collagen type I and III content, with the predominance of the latter, less durable; a decrease in elastin levels and its considerable fragmentation. The greatest expression of tissue degradation was observed among women with GP and CTD manifestations on account of increased MMP-1 and -2 levels; TIMP-1 content was lowest in the group. Associations with GP development have been established among women with CTD signs for genetic polymorphisms: rs3918242 СT gene MMP9 (0,54) (p = 0,007; OR = 3,2; 95% CI 1,3-7,6), rs17576 AG gene MMP9 (0.62 vs. 0,32, p = 0,01; OR = 2,9; 95% CI 1,2-7,0); rs3025058 5A6A gene MMP3 (0.52 vs. 0.45, p = 0.009; OR = 3.7; 95% CI 1,3-10,1); rs2285053 (rs2285052) CT gene MMP2 (0.44 vs. 0.27, p = 0,007; OR = 3,2; 95% CI 1,3-7,5). Statistical significance for the groups was preserved after the correction for multiple comparisons. Summary: the data obtained reveal pathogenetic aspects of genital prolapse – the prevalence of extracellular matrix degradation in a dysplastic morphogenesis. Genetic predictors of pelvic floor remodeling including the formation of its insolvency were established, allowing to extend the range of diagnostic possibilities of the disease progression at early stages or detection the risk of recurrence after surgical treatment. Personification of high-risk groups conducting provides for the exclusion or modification of all the factors predisposing to the development of the disease and performing timely treatment and preventive measures.

Effect of laminarin polysaccharide on activity of matrix metalloproteinase in photoaging skin

To study the effect of laminarin polysaccharide (LP) on the activity of matrix metalloproteinase of photoaging skins. Kunming SPF mice were prepared with back hair shaved, and randomly divided into the control group, the model group, the LP low does group (LP-L, 1 mg x kg(-1)), the LP high dose group (LP-H, 5 mg x kg(-1)) and the Vit E (100 mg x kg(-1)) group. They were abdominally injected with drugs twice on a daily basis. Except for the control group, all groups were exposed to ultraviolet rays for 1 hour every day, five times on a weekly basis, with accumulated exposure dose of UVB being 21.60 J x cm(-2) and accumulated exposure dose of UVA being 84.02 J x cm(-2). Eight weeks later, exposed back skins were collected to detect thickness of dermis by HE stain, content of hydroxyproline (Hyp) by chemical colorimetry, and serum MMP-1 and TIMP-1 content by ELISA. In addition, matrix metalloproteinase-1 (MMP-1) mRNA and relative content of tissue inhibitor of metalloproteinase-1 (TIMP1) mRNA was analyzed with Real-time PCR. Compared with the model group, the LP-H group could significantly increase the thickness of dermis, skin Hyp content and serum TIMP-1 level, and decrease relative content of MMP-1 mRNA in skin and MMP-1 content in serum. LP can regulate the metabolism of collagen photoaging skins by adjusting the activity of matrix metalloproteinase.

Tissue Inhibitors of Matrix Metalloproteinases 1 and 2 and Matrix Metalloproteinase Activity in the Serum and Lungs of Mice with Lewis Lung Carcinoma

We studied the content of tissue inhibitors of matrix metalloproteinases 1 and 2 (TIMP-1 and TIMP-2) and activities of matrix metalloproteinases (MMP) in the serum and lungs of mice with Lewis lung carcinoma metastasizing into the lung. Metastasizing was associated with increased serum content of TIMP-1 and TIMP-2 (only on day 20 at the terminal stage of the tumor process). These data confirm the hypothesis on pro-tumorigenic role of TIMP-1 in the serum. Locally, the development of metastases was associated with a decrease in TIPM-1 concentration (day 7), an increase in TIMP-2 concentration (days 7 and 20), and elevated activity of MMP at all terms of the study (days 7, 15, and 20). Increased concentration of TIMP-2 in the lungs (but not in the serum) can be regarded as an indicator of Lewis lung carcinoma metastasizing.

Menstrual activity of matrix metalloproteinases is decreased in endometrium regenerating after thermal ablation

Menstruation is associated with a striking increase in matrix metalloproteinase (MMP) activity. However, it is still unknown whether the level of MMP activity correlates with the amount of menstrual bleeding. We used histochemistry to investigate the degradation of the extracellular matrix (ECM), and immunohistochemical labelling and zymographic analysis to determine the level of expression and activity of MMP-2 and -9, and of their tissue inhibitors (TIMPs) -1, -2 and -3, in endometria sampled during menstruation in 14 women experiencing excessive menstrual bleeding and in 10 women successfully treated for menorrhagia by thermal ablation of the endometrium. After thermal ablation, regenerated menstrual endometria showed reduced areas of collagen fibre lysis and increased content of TIMP-1 and TIMP-2 compared with endometria from non-treated menorrhagic women. Surprisingly, treated endometria contained more latent gelatinase A (proMMP-2) but a lower proportion of the active form of gelatinase B (MMP-9) than non-treated endometria. These results suggest that ECM degradation is decreased at menstruation in the endometrium regenerated after thermal ablation, mostly because of an increased TIMP expression. This represents the first molecular explanation for the decreased amount of menstrual bleeding.