Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.4.24.35 (matrix metalloproteinase 9)
 2,207 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The literature has revealed variations in the protocols for myoblast cultures, and little information is available on myoblast and fibroblast proliferation. Therefore, the purposes of this study were to establish a prudent protocol for myoblast cultures by comparing a variety of culturing procedures used in previous research and to quantitate myoblast proliferation and fusion under different culture conditions. In addition, the growth status of myoblasts and fibroblasts was investigated. Results indicate that the requirements for an ideal myoblast culture should include a combined enzyme of 0.25% trypsin and 0.2% collagenase type IV (1:1), a preplating time of approximately 15-20 minutes, and a seeding density at 1 x 10(5) cells/ml. Furthermore, the mouse sample should be those of newborns. A better proliferative capacity of myoblasts was noted in an incubator of 10% CO2, coupled with Dulbecco's MEM plus 15% fetal calf serum. The doubling times of myoblasts were shorter than those of fibroblasts, and myoblast number reached its highest at 4 and 5 days. The findings of this study are valuable in understanding the growth status of myoblasts and fibroblasts in primary cultures. Moreover, the establishment of requirements for a good growth of myoblast cultures will facilitate myoblast transfer therapy.
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PMID:Primary culture of mouse myoblasts. 762 15

For diabetes mellitus, little research has been done on the tissue-based or cell-based drug screening model, which has advantages over traditional animal diabetic model in high specificity, high screening volume, low cost and simple manipulation. Considering that the maintenance of complete islet tissue structure is the prerequisite for islet cells to perform their functions normally, an in vitro islet-based drug screening model for diabetes mellitus was established and evaluated. Pancreatic islets were isolated from 3 weeks old mice of either sex by collagenase digestion and density gradient centrifugation as prescribed by Ramanadham S. The volume of 0.1% (W/V) collagenase IV, 0.1% (W/V) Hyaluroridase and 0.1% (W/V) DNase I were 4 times, 2 times and 1 times that of the islets to be digested. And a 2 hours' cold digestion at 4 degrees C was followed by a 10 minutes' warm digestion at 37 degrees C. Under the optimized digestion condition, the islet recovery could be increased by 10%. The isolated islets could survive 6 weeks in vitro and show stable insulin secretion in the first 10 days after inoculation. The obtained islets were cultured in RPMI-1640 medium at 37 degrees C with 5% CO2. Then a diabetic model was established by selecting streptozotocin (STZ) as the evocator and nitric oxide (NO) as the responding index. After 1 day's inoculation, islets culture was treated with STZ, whose concentration ranged from 0 to 5.0 mmol/L. NO was measured by a colorimetric assay at 540nm based on the Griess reaction for 10 min with 0.1 mL Griess reagent and 0.1 mL culture supernatants. Insulin secretion was assayed by RIA methods. Due to the islets-related inoculation variations, NO release and insulin content were both expressed as a percentage of the value recorded in basal experiment which was in the only presence of Krebs culture medium. It was testified that the amount of NO released from islet itself remained steady at 30-35 mmol/L regardless of the changes of STZ concentration from 0 to 5.0 mmol/L. However the NO content in the supernatants of islets culture had close relationship with STZ concentration. This indicated that in this STZ-induced islet diabetic model, NO mainly comes from STZ when it dissolves in water. On the other hand, when STZ changed from 0 to 5.0 mmol/L, the dose-dependent relationship between NO content and insulin secretion showed that the increase of NO came along with the decrease of insulin secretion, which is an important symbol of islet function. As a kind of oxidative free radical, NO is capable of impair islet cells. Thus, NO is a reliable responding index of the model. The optimal STZ concentration in the model is finally determined to be 5.0 mmol/L, under which condition the NO content and insulin secretion is 10.81 times and 0.43 times that in the medium before STZ is added. So if anything is effective in lowering the NO content in the culture, it could protect islets cells from the oxidative attacks of NO. Finally, as an application of the model, the scavenging effect of KOSCr on NO was studied. In a series of KOSCr with different chromium content, all had shown better NO scavenging effects than KOS itself, which could give us an enlightenment of the influence of chromium ion on oligosaccharide. And 1 g/mL KOSCr with 3.519% chromium content can significantly inhibit the NO formation. This has lain a theoretic basis for the research of KOSCr bioactivity and quality control. These results suggested that the STZ-induced diabetic islet model which is impaired by NO free radical can be used effectively, fast and conveniently when screening potential diabetes drugs.
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PMID:[Establishment and application of the model of islet impaired by NO free radical released from streptozotocin]. 1596 20

An abnormal increase in proteolytic enzymes is thought to play a key role in pulmonary emphysema. Alveolar macrophage proteolytic enzymes include cathepsin L, cathepsin S, matrix metalloproteinase 1, 9, and 12, and a number of studies have implicated these proteinases in the alveolar destruction that characterizes emphysema. The aim of this study was to investigate cathepsin L, cathepsin S, matrix metalloproteinase 1, 9, and 12 mRNA expression in alveolar macrophages isolated from patients with varying degrees of emphysema and to correlate their level of expression with measures of emphysema. Alveolar macrophages were isolated from fifty-four patients who underwent surgical resection for lung carcinoma. The level of mRNA expression was determined using real-time PCR. Emphysema was quantified using high-resolution CT scans. Alveolar macrophages were also cultured for 24 h and 48 h; the effect of proinflammatory mediators and promoter polymorphisms on expression was analyzed. There was a significant correlation between matrix metalloproteinase 1 mRNA expression and emphysema. A higher level of matrix metalloproteinase 1 mRNA was associated with more severe emphysema. Matrix metalloproteinase 12 mRNA expression was increased in current smokers as compared with former smokers. Furthermore, there was a negative correlation between matrix metalloproteinase 12 gene expression and carbon monoxide diffusing capacity. The matrix metalloproteinase 9 C-1562T polymorphism significantly influenced matrix metalloproteinase 9 mRNA expression in alveolar macrophages. These results suggest that alveolar macrophage matrix metalloproteinase 1 and 12 may have a role in the lung structural changes leading to the development of emphysema. Furthermore, these data provide evidence to support the concept that multiple proteinases, causing both elastin and collagen degradation, are important in the pathogenesis of pulmonary emphysema.
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PMID:Matrix metalloproteinase expression by human alveolar macrophages in relation to emphysema. 1825 71

Treatment of keloids (K) and hypertrophic scars (HTS) is challenging. A few case reports reported good results in HTS treated by fractional CO2 laser. The aim of the present study was the assessment of the clinical response as well as histological changes in K and HTS treated by fractional CO2 laser and the role of matrix metalloproteinase 9 (MMP9) in the response. A randomized half of the scar was treated by fractional CO2 laser in 30 patients (18 K, 12 HTS) for a total of four sessions 6 weeks apart. Vancouver scar score (VSS) was done before and 1, 3, and 6 months after the last laser session by a blinded observer. Biopsies taken from normal skin, untreated scar, and treated scar tissue 1 and 3 months after the laser sessions were stained by HX & E for histological changes and Masson trichrome for collagen fiber arrangement. Immunohistochemical staining for MMP9 was done in before and 1 month after samples. Quantitative morphometric analysis was done for collagen and MMP9 by image analyzer. Nineteen patients completed the 6-month follow-up period (12 K, 7 HTS). VSS score was significantly lower in the treated compared to untreated areas after 3 and 6 months in both K and HTS but was mainly due to improved pliability of the scar. Histologically, dense inflammatory infiltrate and increased vascularity was apparent 1 month after laser sessions and disappeared at 3 months. Thinner better organized collagen bundle could be seen in 3 months after samples. MMP9 was significantly increased in after treatment samples but without significant correlation with VSS. Fractional CO2 resurfacing is safe but affects mainly pliability of K and HTS with collagen remodeling apparent 3 months after therapy. MMP9 may play a role in mechanism of action of CO2 laser in K and HTS.
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PMID:Treatment of hypertrophic scars and keloids by fractional carbon dioxide laser: a clinical, histological, and immunohistochemical study. 2649 51

Carbonic anhydrase 8 (CA8), a member of the carbonic anhydrase family, is one of the three isozymes that do not catalyze the reversible hydration of carbon dioxide due to the lack of one important histidine. In the present study, we observed increased expression of CA8 in more aggressive types of human osteosarcoma (OS) cells and found that CA8 expression is correlated with disease stages, such that more intense expression occurs in the disease late stage. We also demonstrated that overexpression of CA8 in human OS (HOS) cells significantly increased cell proliferation both in vitro and in vivo. Downregulated CA8 sensitized cells to apoptotic stress induced by staurosporine and cisplatin, suggesting a specific role of CA8 to protect cells from stresses. In addition, downregulation of CA8 in HOS cells reduced cell invasion and colony formation ability in soft agar and further decreased matrix metalloproteinase 9 and focal adhesion kinase expression, indicating that CA8 might facilitate cancer cell invasion via the activation of FAK-MMP9 signaling. Interestingly, HOS cells with CA8 knockdown showed a significant decrease in glycolytic activity and cell death under glucose withdrawal, further indicating that CA8 may be involved in regulating aerobic glycolysis and enhancing cell viability. Knockdown of CA8 significantly decreased phosphorylated Akt expression suggesting that the oncogenic role of CA8 may be mediated by the regulation of Akt activation through p-Akt induction. Importantly, the inhibition of glycolysis by 2-deoxyglucose sensitized CA8 HOS-CA8-myc cells to cisplatin treatment under low glucose condition, highlighting a new therapeutic option for OS cancer.
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PMID:Oncogenic roles of carbonic anhydrase 8 in human osteosarcoma cells. 2671 83

Hypertrophic scarring is a fibroproliferative process that occurs following a third-degree dermal burn injury, producing significant morbidity due to persistent pain, itching, cosmetic disfigurement, and loss of function due to contractures. Ablative fractional lasers have emerged clinically as a fundamental or standard therapeutic modality for hypertrophic burn scars. Yet the examination of their histopathological and biochemical mechanisms of tissue remodeling and comparison among different laser types has been lacking. In addition, deficiency of a relevant animal model limits our ability to gain a better understanding of hypertrophic scar pathophysiology. To evaluate the effect of ablative fractional lasers on hypertrophic third-degree burn scars, we have developed an in vivo Red Duroc porcine model. Third-degree burn wounds were created on the backs of animals, and burn scars were allowed to develop for 70 days before treatment. Scars received treatment with either CO2 or erbium: yttrium aluminum garnet (YAG) ablative fractional lasers. Here, we describe the effect of both lasers on hypertrophic third-degree burn scars in Red Duroc pigs. In this report, we found that Er:YAG has improved outcomes versus fractional CO2. Molecular changes noted in the areas of dermal remodeling indicated that matrix metalloproteinase 2, matrix metalloproteinase 9, and Decorin may play a role in this dermal remodeling and account for the enhanced effect of the Er:YAG laser. We have demonstrated that ablative fractional laser treatment of burn scars can lead to favorable clinical, histological, and molecular changes. This study provides support that hypertrophic third-degree burn scars can be modified by fractional laser treatment.
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PMID:Assessment of Ablative Fractional CO2 Laser and Er:YAG Laser to Treat Hypertrophic Scars in a Red Duroc Pig Model. 2971 76