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OBJECTIVE@#To explore the effect of miR-143 regulating matrix metalloproteinase(MMP)-13 expression on migration and invasion of osteosarcoma cells.@*METHODS@#The mouse osteosarcoma cell line 143B cells were cultured in 96-well plates, and blank group, negative group, positive group, and intervention group were set up. Then, the blank group did no treatment 50 μg miR-143 mimic was added to positive group, negative group added equal mimic NC (control sequence of miR-143 mimic), the intervention group was added 50 μg miR-143 mimic and 10 μg MMP-13 protein, all groups continued to culture for 3 to 6 hours, and finally the serum was aspirated to treat for half an hour. The protein expressions of miR-143 and MMP-13 in each group were measured by fluorescence quantitative PCR experiment and Western blot experiment, respectively, and the invasion and migration abilities of cells were measured by Transwell and scratch experiments.@*RESULTS@#The expression of MMP-13 protein in the positive group and the intervention group was significantly lower than that in the blank group, and the positive group was lower than the intervention group (P<0.05);The mean numbers of invasive cells in blank group, negative group, positive group and intervention group were (1 000.01±44.77), (959.25±46.32), (245.04±4.33), (634.06±33.78) cells/field, respectively;the scratch healing rate of the positive group and the intervention group was significantly lower than that of the blank group, and the positive group was lower than the intervention group (P<0.05).@*CONCLUSION@#MMP-13 is a target of miR-143, which can reduce the migration and invasion ability of osteosarcoma cells by inhibiting the expression of MMP-13.
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Animals , Mice , Osteosarcoma/pathology , MicroRNAs/genetics , Matrix Metalloproteinase 13/genetics , Neoplasm Invasiveness , Cell Line, Tumor , Cell MovementABSTRACT
Objective: inflammation may play a role in bone loss by altering the boné remodelling process, favouring bone resorption by osteoclasts over bone synthesis by osteoblasts. Matrix metalloproteinase 13 (MMP-13) has the ability to activate osteoclasts, leading to bone resorption. Regenerative treatments have been widely used in periodontology. When combined with Platelet-rich fibrin (PRF), xenografts will give better results in bone regeneration. The aim of this study was to evaluate the effect of xenograft combined with PRF on MMP-13 expression in a bone defect using an experimentally created bone defect. Material and Methods: eighteen New Zealand rabbits were assigned to three groups. Each group consisted of six New Zealand rabbits. A critical bone defect with a diameter size of 5 mm was created in the right tibia of each rabbit in group 1 (application: xenograft), group 2 (application: PRF), and group 3 (application: xenograft and PRF). The PRF was produced from 5 ml of blood taken from each rabbit's ears. After 30 days, the rabbits were euthanized. The tissue samples were evaluated by immunohistochemical staining. Results: group 3 showed the lowest mean expression of MMP-13 (4.50) compared to group 1 (20.50) and group 2 (11.70). Group 3 showed a significant difference in the MMP-13 expression compared to group 1 and group 2 (P = 0.000) (P < 0.05). Conclusion: this research showed that the combination of xenograft and PRF had the lowest expression of MMP-13. The application of a xenograft and PRF has better osteogenesis ability in bone regeneration.(AU)
Objetivo: inflamação pode interferir na perda óssea através de alterações no processo de remodelação, favorecendo a reabsorção óssea pelos osteoclastos ao invés da síntese pelos osteoblastos. A metaloproteinases de matriz 13 (MMP-13) ativa osteoclastos causando reabsorção óssea. Tratamentos regenerativos têm sido amplamente usados na periodontia. Quando combinamos Plasma rico em plaquetas (PRP) e xenoenxerto levam a melhores resultados de regeneração óssea. O objetivo deste estudo foi avaliar os efeitos de xenoenxerto combinado com PRP na expressão de MMP-13 em defeitos ósseos experimentais. Material e Métodos: dezoitos coelhos Nova Zelândia foram distribuídos em 3 grupos de 6 coelhos cada. Um defeito ósseo de 5 mm de diâmetro foi feito na tíbia direita dos animais do grupo 1 (xenoenxerto), grupo 2 (PRP) e grupo 3 (Xenoenxerto+PRP). O PRP foi obtido pela coleta de 5mL de sangue das orelhas dos coelhos. Após 30 dias, os coelhos foram eutanasiados. As amostras foram submetidas a coloração imuno-histoquímica. Resultados: o grupo 3 apresentou a menor expressão de MMP-13 (4.50) quando comparado ao grupo 1 (20.50) e ao grupo 2 (11.70). O grupo 3 mostrou diferença estatística significante em relação a expressão de MMP-13 quando comparado aos grupos 1 e 2 (p=0.000) (p< 0.05). Conclusão: esta pesquisa mostra que a combinação de xenoenxerto e PRP teve a menor expressão de MMP-13. A combinação de xenoenxerto e PRP têm maior habilidade de osteogênese na regeneração óssea (AU)
Subject(s)
Animals , Rabbits , Bone Regeneration , Platelet-Rich Plasma , Matrix Metalloproteinase 13 , Heterografts , InflammationABSTRACT
Objective: To detect the expression of MMP-13 in the skin of mice treated by near-infrared (IRA) and ultraviolet (UV) so as to observe their effects on skin photoaging and their interrelation and explore molecular mechanisms in IRA-induced skin photoaging. Methods: Male ICR mice were randomly divided into five groups: control group (without ray exposure), IRA group, UV group, IRA/UV group, and UV/IRA group. The mice in the latter four groups had their dorsal skin exposed to different radiated ray respectively. The levels of MMP-13 protein and mRNA in the exposed skin were detected by HE, immunohistochemical method, Western blot and real-time PCR. Results: Both skin lesions by visual inspection and H&E staining results showed that mice in the other four groups had skin photoaging in the exposed skin area compared with the control group. The levels of MMP-13 protein and mRNA in the exposed skin in IRA/UV and UV/IRA groups were significantly higher than those in the control mice (P<0.05). In addition, mice in IRA/UV group showed higher levels of MMP-13 protein and mRNA than those in UV/IRA group (P<0.05). Conclusion: ① IRA causes skin photoaging in mice. ② UV and IRA interact with each other, up-regulate the expression of MMP-13, and promote each other in the process of photoaging. ③ The effects of IRA and UV in combination on skin photoaging are closely related to order of exposure. Taken together, avoiding IRA exposure and the expression of MMP-13 play an important role in preventing skin wrinkle formation and treatment of photoaging in mice.
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Objective: To observe the effects of He-Ne laser on the expression of extracellular matrix and its regulatory factors in degenerated temporomandibular joint(TMJ) of rabbits. Methods: 40 New Zealand adult white rabbits were randomly divided into normal group,sham model group,TMJOA model group and laser treatment group(n = 10),the rats in treatment group were treated by He-Ne laser irradination at acupoints. The rats in each group were divided into 1 d and 11 d groups(n = 5). The animals were respectively killed 1 d and 11 d after operation and HE staining was used to observe the histomorpholy. The protein expression of extracellular matrix and its regulatory factors were examined by Western blot. Results: After He-Ne laser treatment,the fiber layer of condylar cartilage was slightly loose,part of the fiber was newly produced. The level of Col-2,PRG-4,TIMP-1,BMP-2 was up-regulated and the of MMP-13 was down-regulated in the 11 d treatment group. Conclusion: He-Ne laser irradiation on acupoints may up-regulate the expression of extracellular matrix (Col-2 and PRG-4) and its regulatory factors (TIMP-1 and BMP-2),down-regulate the expression of MMP-13.
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Arthritis is a common chronic disease characterized by the destruction of joint cartilage and inflammation in the sur-rounding tissues. Although it is known that the pathogenesis of arthritis is influenced by a series of factors, the underlying mechanisms remain unclarified. Recently, increasing attention has been paid to the increase of matrix metalloproteinases (MMPs) in articular cartilage,resulting in an inevitable degra-dation of cartilage and extracellular matrix (ECM). MMP-13, the major functioning enzyme during arthritis development,plays a vital role in the cartilage destruction, thus contributing to the decomposition of type Ⅱ collagen irreversibly. A variety of cellu-lar cytokines such as IL-1β and TNF-α,and Runx2 are assumed to affect the expression of MMP-13 in chondrocytes. The hypom-ethylation of the promoter region of the MMP-13 may induce its expression, while it can be reduced by inhibiting histone acety-lation. Meanwhile, microRNA can reduce the expression of MMP-13. In conclusion, MMP-13 can be used as an important therapeutic target in arthritis. In this review, we focus on the role of MMP-13 in arthritis and its underlying regulatory mecha-nisms.
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Objective To investigate the feasibility of osteoarthritis (OA) model in rabbits by injuring medial meniscus so as to understand the role of interleukin-1 beta (IL-1β) and matrix metalloproteinase-13 (MMP-13) in pathological mechanism of articular cartilage and synovial fluid of OA model in rabbits.Methods We randomly divided 40 New Zealand white rabbits into experimental group (n =30) and control group (n =10).Pathological changes in articular cartilage of the femoral condyle were scored at weeks 2,6 and 12 after surgery.We detected the expressions of IL-1β and MMP-13 by immunohistochemistry.The cell fractions of IL-1β and MMP-13 were recorded by ELISA.Results The articular cartilage score and HE staining significantly differed at various time points of gross and pathological observation between control group and experimental group (P＜ 0.05).Immunohistochemistry showed that IL-1β was expressed in both groups and that the cell fraction differed significantly at weeks 6 and 12 (P＜0.05),but not at week 2 between the two groups.MMP-13 protein expression was not detected in articular condrocytes in control group,but was detected in experiment group with a significant difference (P＜0.05).IL-1β expression was consistent in articular cartilage and synovial fluid.Conclusion The reasonable rabbit animal OA model could be established by knee meniscus injury caused by surgical method.Expressions of IL-1β and MMP-13 change obviously in the pathomechanism of OA.Further clinical studies are needed to determine whether they can be used as markers in early diagnosis of OA.
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Objective:To analysis the correlation of Wnt/β-catenin signaling pathway,OPN and MMP-13 in rabbit model with different degree of osteoarthritis (OA).Methods:Forty New Zealand white rabbits were randomly divided into four groups of 10 each,then we constructed the rabbit model with different degree of osteoarthritis (OA) by different concentrations of papain.The mRNA and protein levels of β-catenin,OPN,MMP-13,type Ⅱ collagen and proteoglycan were detected by Real-Time PCR and ELISA,respectively.Results:We have successfully established OA rabbit model by different concentrations of papain,and these OA rabbits model could be divided into mild,moderate and severe three level by Mankin scoring system,chondrocytes.Compared with the normal control group,the protein levels of β-catenin,OPN,and MMP-13 in the osteoarthritis were significantly increased,while the protein levels of type Ⅱ collagen and proteoglycan were significantly decreased (P＜0.05).With the increasing severity of OA,the results were consistent (P＜0.05).Conclusions:These results indicated that Wnt/β-catenin signaling pathway might regulate the expression of OPN,thereby affecting the expression of MMP-13,and ultimately have an impact on the occurrence ofosteoarthritis.
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Objective To study the effects of supplemental Dangguibeimukushen pill prescription on the expressions of MMP13 and bFGF in tumor tissues on H22 hepatoma bearing mice.Methods H22 hepatoma bearing mice was employed to carry out anti-tumor experiment in vivo in this study.A total of 60 mice were randomly divided into model group,DDP positive control group,low and high dosage of supplemental Dangguibeimukushen pill treatment group,supplemental Dangguibeimukushen pill combined with DDP group.In each gruop,gastric infusion was performed continuously 14 d.The tumor growth and the general conditions of mice were recorded.After the last administration of gavages orally treatment,all mice were anaesthetized and killed by cervical disloca-tion method to obtain completely tumor tissue for further HE staining measure and detection of MMP13 and bFGF in tumor tissue with the method of RT-qPCR and immunohistochemistry.Results The tumor volume of model group and low dosage of supple-mental Dangguibeimukushen pill treatment group were larger than that of other groups,futhermore,ulceration were found inside tumor,and basic integrity were observed of tumor capsule.However,in all the treatment groups,the tumor volume was relatively smaller,and the capsule of tumor was incomplete,ulceration of different degrees and bleeding when tumor was peeled off.In combi-nation groups,the tumor volume was much smaller,and the tumor had incomplete capsule and less ulceration inside.Pathological observation showed that connective tissue was found inside tumor,cells were sparsely spread and the number of necrosis cancer cells were increased in all treatment groups.RT-qPCR detection showed that the mRNA expression of MMP13 and bFGF in each treat-ment group were decreased comparing with the model group,and the difference was statistically significant(P <0.05).Immunohis-tochemical detection showed that the expression of MMP13 and bFGF in the combined group were less colored and weakly positive expression comparing with the positive group or the pure Chinese medicine treatment group,the difference was statistically signifi-cant(P <0.05).Conclusion Dangguibeimukushen pill could downregulate the expression of MMP13 and bFGF of tumor tissue in H22 hepatocarcinoma xenograft mice at mRNA and protein levels,and played a vital role in inhibiting cancer invasion and angiogen-esis,reducing toxics and side effects and enchancing its efficiency.
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Purpose To analyze the MMP-9,MMP-13,HIF-1α expression in lung adenocarcinoma tissue and to explore the relationship with clinical pathologic features,EGFR mutation and prognosis of the patients.Methods The expression of MMP-9,MMP-13,HIF-1α in 629 cases of lung adenocarcinoma were detected by using immunohistochemical of SP method.50 cases of normal tissue adjacent to carcinoma and 50 cases of pneumonia pseudotumor hyperplasia tissues were selected as controls.629 patients with lung adenocarcinoma were detected by real-time fluorescence quantitative PCR and all of them were followed-up.Results The MMP-9,MMP-13,HIF-1α expression in lung adenocarcinoma tissues were higher than controls (P ＜ 0.001).The MMP-9 expression was correlated with lymph node metastasis and the size of the tumor (P ＜ 0.05).The MMP-13 expression was correlated with smoking history,TNM stage,lymph node metastasis and the size of the tumor (P ＜0.05).The HIF-1α expression were correlated with smoking and lymph node metastasis.Statistically significant differences were found in all these above groups (P ＜ 0.05).But the expression of all has nothing to do with the EGFR mutation (P ＞0.05).The Kaplan-Meier survival analysis results showed that MMP-9,MMP-13,HIF-1α positive expression groups of 12,24 and 36 months cumulative survival were significantly lower than the negative expression groups.COX multiple factor analysis results showed that EGFR mutation,the expression of MMP-9,MMP-13,HIF-1α,tumor size,TNM stage were independent risk factors for the development of lung adenocarcinoma patients living conditions.Statistically significant differences were found in these groups (P ＜0.05).Conclusion MMP-9,MMP-13,HIF-1α are overexpressed in lung adenocarcinoma tissues,and its expression are associated with lymph node metastasis,but has nothing to do with EGFR mutations.Patients with positive expression of MMP-9,MMP-13,HIF-1α and with no EGFR mutation have lower survival rates,and they are independent risk factors for the development of lung adenocarcinoma patients living conditions.
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Objective To observe the effects of intra-articular injection of sinomenine on morphology of synovium and cartilage as well as contents of MMP-13 and cartilage oligomeric matrix protein(COMP) in serum and synovial fluid of rabbit knee osteoarthritis model.Methods A total of 38 New Zealand white rabbits were randomly divided into 4 groups: control group, model group, hyaluronate group and sinomenine group. Model group, hyaluronate group and sinomenine group established knee osteoarthritis model by intra-articular injection of papain enzyme, and were treated with saline, sodium hyaluronate and sinomenine respectively except for control group. 5 weeks after treatment, all rabbits were sacrificed for HE staining and histological grading on cartilago articularis and synovium, and ELISA method was used to detect the contents of MMP-13 and COMP in rabbit serum and synovial fluid.Results Mankin's scores of articular cartilage and histological scores of synovium in model group were significantly higher than those of control group (P<0.01), and were markedly lower in sinomenine group than those of model group (P<0.01); the contents of MMP-13 and COMP in serum and synovial fluid of model group were significantly raised compared with control group (P<0.01), and were reduced obviously in sinomenine group compared with model group (P<0.05,P<0.01).Conclusion Intra-articular injection of sinomenine can reduce the levels of MMP-13 and COMP in serum and synovial fluid of rabbit knee osteoarthritis model, and improve synovial inflammation, as well as delay the degradation of articular cartilage.
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<p><b>OBJECTIVE</b>To observe the effects of close-to-bone needing combined with electroacupuncture (EA) on cartilage collagen type Ⅱ/discoidin domain receptor/matrix metalloproteinase 13 (collagen type Ⅱ/DDR2/MMP 13) signaling pathway in rabbits with knee osteoarthritis (KOA), and to explore the possible action mechanism of this method on repair of extracellular matrix of knee cartilage.</p><p><b>METHODS</b>Forty New Zealand white rabbits were randomly assigned into a normal group (10 rabbits) and a model establishing group (30 rabbits). The Hulth-Telhag technique was applied to establish the model of KOA, and X-ray was used for outcome assessment. The rabbits with successful modeling were randomly assigned into a model group, a close-to-bone needing group, a regular acupuncture group, 10 rabbits in each one. The rabbits in the close-to-bone needing group were treated with close-to-bone needing and EA; the rabbits in the regular acupuncture group were treated with regular acupuncture and EA. "Neixiyan" (EX-LE 4), "Dubi" (ST 35), "Yinlingquan" (SP 9), "Zusanli" (ST 36) and "Liangqiu" (ST 34) were selected in the two groups. The intervention was given for 20 min, once a day; the intervention of 5 days made 1 session, 2 days as the interval and totally 4 sessions were given. Rabbits in normal and model group were immobilized without any treatment. After the treatment, western blotting method was applied to evaluate the expression of DDR2 and collagen type Ⅱ; the activity of collagen type Ⅱ, DDR2 and MMP 13 was assessed by immunohistochemistry method; the mRNA expression of DDR2 and MMP 13 was determined by reverse transcription-polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>Compared with the normal group, the activity expression of collagen type Ⅱ were significantly reduced in the other groups (all<0.01),while the activity and mRNA expression of DDR2 and MMP 13 were notably increased (all<0.01). Compared with the model group, the activity expression of collagen type Ⅱ in the close-to-bone needing group and regular acupuncture group were increased (all<0.01), while the activity and mRNA expression of DDR2 and MMP 13 were reduced (all<0.01). Compared with the regular acupuncture group, the activity and mRNA expression of MMP 13 and DDR2 in the close-to-bone needing group were reduced (all<0.01), while the activity expression of collagen type Ⅱ were increased (<0.01).</p><p><b>CONCLUSIONS</b>The close-to-bone needing combined with EA and regular EA could both promote the repair of knee cartilage, where closing-to-bone needing combined with EA shows a superior efficacy. The mechanism may be associated with the blocking effect of collagen type Ⅱ/DDR2/MMP13 signaling pathway and the inhibiting effect of degradation in extracellular matrix of cartilage.</p>
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We aimed to investigate the effects of an anti-tumor necrosis factor-α antibody (ATNF) on cartilage and subchondral bone in a rat model of osteoarthritis. Twenty-four rats were randomly divided into three groups: sham-operated group (n=8); anterior cruciate ligament transection (ACLT)+normal saline (NS) group (n=8); and ACLT+ATNF group (n=8). The rats in the ACLT+ATNF group received subcutaneous injections of ATNF (20 μg/kg) for 12 weeks, while those in the ACLT+NS group received NS at the same dose for 12 weeks. All rats were euthanized at 12 weeks after surgery and specimens from the affected knees were harvested. Hematoxylin and eosin staining, Masson's trichrome staining, and Mankin score assessment were carried out to evaluate the cartilage status and cartilage matrix degradation. Matrix metalloproteinase (MMP)-13 immunohistochemistry was performed to assess the cartilage molecular metabolism. Bone histomorphometry was used to observe the subchondral trabecular microstructure. Compared with the rats in the ACLT+NS group, histological and Mankin score analyses showed that ATNF treatment reduced the severity of the cartilage lesions and led to a lower Mankin score. Immunohistochemical and histomorphometric analyses revealed that ATNF treatment reduced the ACLT-induced destruction of the subchondral trabecular microstructure, and decreased MMP-13 expression. ATNF treatment may delay degradation of the extracellular matrix via a decrease in MMP-13 expression. ATNF treatment probably protects articular cartilage by improving the structure of the subchondral bone and reducing the degradation of the cartilage matrix.
Subject(s)
Animals , Female , Adalimumab/pharmacology , Antirheumatic Agents/pharmacology , Bone and Bones/drug effects , Cartilage, Articular/drug effects , Osteoarthritis/drug therapy , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Arthroplasty, Subchondral , Anterior Cruciate Ligament/surgery , Arthritis, Experimental/drug therapy , Bone and Bones/metabolism , Cartilage, Articular/metabolism , Extracellular Matrix/drug effects , Hindlimb/pathology , Hindlimb/surgery , Immunohistochemistry , Injury Severity Score , /drug effects , /metabolism , Osteoarthritis/surgery , Protective Factors , Random Allocation , Rats, Sprague-DawleyABSTRACT
OBJECTIVE@#To study the effect of low intensity pulsed ultrasound (LIPUS) on the expression of tissue inhibitor of metalloproteinase-2 (TIMP-2) in the serum and expression of matrix metallopeptidase 13 (MMP-13) in the articular cartilage cells of rabbits with knee osteoarthritis (OA).@*METHODS@#Inner patellar ligament defect method was used to establish the model of knee OA. Four weeks after the modeling, the arterial blood was drawn from the ear of each rabbit, while ELISA was employed to detect the expression of TIMP-2 in the serum. The chondrocytes were separated from animals in each group and then cultured in vitro. All rabbits were divided into control group, OA model group and OA + LIPUS group. Cells in the control and OA groups were not treated, while cells in the OA + LIPUS group were treated with LIPUS (40 mW/cm(2), 1 time/day). Cells were collected 7 d later and the RNA and total protein were extracted respectively. Real-time PCR and Western blotting were employed to analyze the expression of MMP-13 in chondrocytes at the mRNA and protein level, respectively.@*RESULTS@#The success rate of establishment of OA model was 83%. The results of ELISA showed that the content of TIMP-2 in the serum of animals with OA was 22.3%, lower than the one in the control group (P < 0.05). Compared with the normal control group, the expression of TIMP-2 in the OA model group was significantly increased, while the expression of MMP-13 was significantly increased (P < 0.05). After the stimulation of LIPUS, the expression of TIMP-2 and MMP-13 was close to the one in the normal control group.@*CONCLUSIONS@#The inner patellar ligament defect method is a mature method to establish the rabbit OA model, with high success rate. The expression of serum TIMP-2 in the OA model group is significantly decreased. LIPUS can up-regulate TIMP-2 and down-regulate MMP-13.
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Objective: To study the effect of low intensity pulsed ultrasound (LIPUS) on the expression of tissue inhibitor of metalloproteinase-2 (TIMP-2) in the serum and expression of matrix metallopeptidase 13 (MMP-13) in the articular cartilage cells of rabbits with knee osteoarthritis (OA). Methods: Inner patellar ligament defect method was used to establish the model of knee OA. Four weeks after the modeling, the arterial blood was drawn from the ear of each rabbit, while ELISA was employed to detect the expression of TIMP-2 in the serum. The chondrocytes were separated from animals in each group and then cultured in vitro. All rabbits were divided into control group, OA model group and OA + LIPUS group. Cells in the control and OA groups were not treated, while cells in the OA + LIPUS group were treated with LIPUS (40 mW/cm
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Objective To investigate the expression levels of Interleukin(IL)?6 and matrix metalloproteinases(MMP)?13 in synovial fluid by in?vigorating kidney and activating blood formulae in treating rabbit knee osteoarthritis model. Methods A total of 30 New Zealand rabbit were ran?domly divided into blank group,model group,invigorating kidney group,activating blood group and invigorating kidney and activating blood group. Rabbits model with knee osteoarthritis were established by improved Hulth method. To give corresponding respectively the medicinal broth,model group was given saline,knee joint synovial fluid was collected after 4,8 and 12 weeks. Enzyme linked immunosorbent assay(ELISA)was used to measure the levels of IL?6 and MMP?13. Results The levels of IL?6 in rabbit knee osteoarthritis were obviously higher than that of normal control group at both 4 weeks and 8 weeks(P<0.001). But there was no statistical difference on the levels of IL?6 compared with controls in 12 weeks. In addition,the level of MMP?13 at 4 weeks,8 weeks and 12 weeks were significantly higher than the blank control group(P<0.001). After 8 weeks of Chinese medicine administration,the levels of IL?6 in synovial fluid were significantly decreased in invigorating kidney group,activating blood group and invigorating kidney and activating blood group(P<0.001),but there was no statistical difference among groups in 12 weeks. The MMP?13 levels of synovial fluid was significantly lower than the model group(P<0.001). Conclusion Our results indicate that IL?6 and MMP?13 par?ticipate in the pathological development of the rabbit knee osteoarthritis. Invigorating kidney and activating blood formulae could reduce the expres?sion of IL?6 and MMP?13 and alleviate osteoarthritis progression,and which is superior to the pure invigorating kidney formulae and activating blood formulae.
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Purpose To explore the role of expression of MMP-9 and MMP-13 in non-small cell lung cancer ( NSCLC) , and to investi-gate their association with the clinicopathologic feactures and prognosis of NSCLC. Methods The expression of MMP-9 and MMP-13 was detected in 88 NSCLC tissues and 18 adjacent normal tissues by immunohistochemistry SP method. Results The expression of MMP-9 and MMP-13 was higher in NSCLC than that in adjacent normal tissues (P<0. 05), which was positively correlated with dif-ferentiation and lymph node metastasis (P<0. 05). The expression of MMP-9 was positively correlated with age (P<0. 05), which was positively correlated with the expression of MMP-13 in NSCLC ( P <0. 05 ) . The survival rates in positive expression group of MMP-9 and MMP-13 were significantly lower than that of the negative expression group by log-rank method comparing survival curves (P<0. 05). Cox model analysis showed that the tumor size, lymph node metastasis and MMP-13 positive expression were closely relat-ed with the prognosis of NSCLC (P<0. 05). Conclusion MMP-9 and MMP-13 both are associated with the metastasis, invasion and prognosis of NSCLC, and MMP-13 may mainly activate MMP-9 to participate the invasion and metastasis of NSCLC.
ABSTRACT
Os miofibroblastos são células que apresentam um fenótipo híbrido exibindo características morfológicas de fibroblastos e de células musculares lisas, sendo a aquisição de tal fenótipo denominada diferenciação, passando então a expressar a a-SMA, a qual é importante na identificação dessas células. Estudos têm sugerido que os miofibrobíastos apresentam relação com a agressividade de diversas lesões e que o seu processo de diferenciação estaria relacionado à expressão do TGF-pl e do IFN-y atuando, respectivamente, no estímulo e na inibição dessa diferenciação. O objetivo deste trabalho foi investigar o papel dos miofibroblastos em lesões odontogênicas epiteliais, relacionando-os à agressividade das lesões e analisar por meio da imuno-histoquímica. a expressão do TGF-pl e IFN-y no processo de diferenciação, além da análise da MMP-13 que é ativada por miofibroblastos e do indutor de metaloproteinases de matriz (EMMPRIN) como precursor desta MMP. A amostra foi constituída por 20 ameloblastomas sólidos, 10 ameloblastomas unicfsticos, 20 ceratocistos odontogênicos e 20 tumores odontogênícos adenomatóides. Para a avaliação dos miofibroblastos, foram quantificadas as células imunorreativas ao anticorpo a-SMA presentes no tecido conjuntivo, próximo ao tecido epitelial. As expressões de TGF-pl, IFN-y, MMP-13 e EMMPRIN, foram avaliadas no componente epitelial e no conjuntivo, estabelecendo-se o percentual de imunorreatividade e atribuindo-se escores de 0 a 4. A análise dos miofibroblastos evidenciou maior concentração nos ameloblastomas sólidos (média de 30,55), seguido pelos ceratocistos odontogênicos (22,50), ameloblastomas unicísticos (20,80) e tumores odontogênicos adenomatóides (19,15) com valor de p= 0,001. Não foi encontrada correlação significativa entre TGF-pl e IFN-y no processo de diferenciação dos miofibroblastos, bem como na relação entre a quantidade de miofibroblastos e a expressão da MMP-13. Constatou-se, correlação estatística entre MMP-13 e TGF-pi (r= 0,087; p= 0,011) além de significante correlação entre MMP-13 e IFN-y (r=0,348; p=0,003). Entre EMMPRÍN e MMP-13 verificou-se significância (r= 0,474; p<0,001) assim como entre EMMPRIN e IFN-y (r=0,393; p=0,001). A maior quantidade de miofibroblastos evidenciada nos ameloblastomas sólidos, ceratocistos odontogênicos e ameloblastomas unicísticos sugere que estas células podem ser um dos fatores responsáveis para um comportamento biológico mais agressivo destas lesões, embora a população de miofibroblastos não tenha apresentado correlação com TGF- -pi, IFN-y ,MMP-13 e EMMPRIN. Quanto a correlação evidenciada entre MMP-13 e TGF-pl, isto pode sugerir um papel indutor do TGF-pl para a expressão da MMP-13, assim como os resultados deste estudo reforçam a relação bem estabelecida do EMMPRIN como indutor da MMP-13. Constatou-se também relação entre EMMPRIN e IFN-y assim como entre MMP-13 e IFN-y sugerindo, dessa forma, um sinergismo na ação anti-fibrótica desses marcadores.
Myofibroblasts are cells that exhibit a hybrid phenotype, sharing the morphoíogical characteristics of fibroblasts and smooth muscle cells, which is acquired during a process called differentiation. These cells then start to express a-SMA, a marker that can be used for their identification. Studies suggest that myofibroblasts are related to the aggressiveness of different tumors and that TGF-pl and IFN-y play a role in myofibroblast differentiation, stimulating or inhibiting this differentiation, respectively. The objective of this study was to investigate the role of myofibroblasts in epithelial odontogenic tumors, correlating the presence of these cells with the aggressiveness of the tumor. Immunohistochemistry was used to evaluate the expression of TGF-pl and IFN-y in myofibroblast differentiation, as well as the expression of MMP-13, which is activated by myofibroblasts, and of EMMPRIN (extracellular matrix metalloproteinase inducer) as a precursor of this MMP. The sample consisted of 20 solid ameloblastomas, 10 unicystic ameloblastomas, 20 odontogenic keratocysts, and 20 adenomatoid odontogenic tumors. For evaluation of myofibroblasts, anti-a-SMA-immunoreactive cells were quantified in connective tissue close to the epithelium. Immunoexpression of TGF-pl, IFN-y, MMP-13 and EMMPRIN was evaluaíed in the epithelial and connective tissue components, attributing scores of 0 to 4. The results showed a higher concentration of myofibroblasts in solid ameloblastomas (mean of 30.55), followed by odontogenic keratocysts (22.50), unicystic ameloblastomas (20.80), and adenomatoid odontogenic tumors (19.15) (p=0.00). No significant correlation between TGF-pl and IFN-y was observed during the process of myofibroblast differentiation. There was also no correlation between the quantity of myofibroblasts and MMP-13 expression. Significant correlations were found between MMP-13 and TGF-pi (r=0.087; p=0.01 1), between MMP-13 and ÍFN-y (r=0.348; p=0.003), as well as between EMMPRIN and MMP-13 (r=0.474; /xO.001) and between EMMPRIN and IFN-y (r=0.393; p=0.00). The higher quantity of myofibroblasts observed in solid ameloblastomas, odontogenic keratocysts and unicystic ameloblastomas suggests that these cells are one of the factors responsible for the more aggressive biological behavior of these tumors, although the myofibroblast population was not correlated with TGF-01, IFN-y, MMP-13 or EMMPRIN. The correlation between MMP-13 and TGF-pl suggests that the latter induces the expression of this metalloproteinase. The present results also support the well-established role of EMMPRIN as an inducer of MMP-13. Furthermore, the relationship between EMMPRIN and IFN-y and between MMP-13 and IFN-y suggests synergism in the antifibrotic effect of these markers.
Subject(s)
Ameloblastoma/pathology , Odontogenic Cysts/etiology , Odontogenic Cysts/pathology , Extracellular Matrix/pathology , Myofibroblasts/physiology , Myofibroblasts/pathology , Transforming Growth Factors , Odontogenic Tumor, Squamous/diagnosis , Odontogenic Tumor, Squamous/pathology , Immunohistochemistry , Statistics, NonparametricABSTRACT
Objective To observe the in vitro effect of leptin, alone or in combination with tumor necrosis factor-alpha (TNF-α) on inducible nitric oxide (NO) and on inducible matrix metallo-proteinase-1 3 (MMP-13) in rabbit articular chondrocytes. Methods The chondrocytes from the articular cartilage of 2-month-old rabbits were cultivated and identified, and the second filial generation chondrocytes were cocultured on plates with different concentrations of leptin alone or in combination with TNF-α for 48 h or 96 h after 12 h starvation. The concentration of NO and MMP-13 was measured in the chondrocytes culture supernatant fluid. The results were statistically analyzed. Results There was no significant difference in the concentrations of NO between the different concentrations of leptin alone groups and the blank control group (P ＞ 0. 05). In combination with the same concentration of TNF-α (10 ng/mL), leptin could dose-dependently increase the concentration of NO in the chondrocytes culture supernatant fluid in vitro. There was significant value in average concentration of MMP-13 on the main effect of both time and dose (P ＜0. 05) . No MMP-13 was detected in the blank control group. Conclusion Leptin can induce MMP-13 and have synergistic induction effect on NO with TNF-α in rabbit articular chondrocytes in vitro.
ABSTRACT
PURPOSE: This study investigated the effect of COX-2 inhibitor on the expression of MMP-13 in the healing process of fracture. MATERIAL AND METHODS: Adult Sprague-Dawley rats were divided into two groups of twenty five rats each. Unilateral femoral shaft fractures were created artificially under displacement in all two groups. COX-2 inhibitor was only given to the experimental group from the postoperative day 1. At 2 weeks after fracture the rats were sacrificed and the callus from each group was used for histologic examination and real time RT-PCR for MMP-13 expression. RESULTS: Histologically, proliferation of osteoblasts and formation of osteoid was less abundant in the experimental group. In real time RT-PCR, the mean expression of MMP-13 is 2.84+/-2.50 in the control group compared with 1.16+/-1.05 in the experimental group. CONCLUSION: In the early stage of fracture healing, COX-2 inhibitor suppress the expression of MMP-13.
Subject(s)
Adult , Animals , Humans , Rats , Bony Callus , Displacement, Psychological , Fracture Healing , Osteoblasts , Rats, Sprague-DawleyABSTRACT
Summary: The aim of this study was to investigate the effect and possible mechanism of all-trans retinoic acid (ATRA) on liver fibrosis induced by common bile duct ligation (CBDL) in rats. Fifty-three female Wistar rats were randomly divided into 5 groups: sham operation group (group J, 5 animals) and groups A, B, C and D (12 animals in each group). The rats in groups A, B, C and D were subjected to CBDL to induce liver fibrosis, while those in group J to sham operation. From the 3rd week the rats in groups B, C and D respectively received daily administration of ATRA via gastric tube at three different doses [0.1, 1.5 and 7.5 mg/kg body weight (BW)]. Animals were sacrificed at 6th week. Rats' liver tissues were observed for pathologic changes under a light microscope. The protein levels of type Ⅰ collagen (COL Ⅰ), matrix metalloproteinase-2 (MMP2), MMP13 and tissue inhibitors of metalloproteinase-1 (TIMP-1) in liver tissues were determined by immunohistochemical techniques. The expression levels of TGF-β1 and CTGF mRNA in liver tissues were detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The results showed that loss of normal hepatic architecture and formation of obvious fibrosis were observed in group A, while ATRA treatment for 4 weeks notably alleviated the pathological changes of hepatocytes. The expression of COL Ⅰ and TIMP-1 proteins in group A was increased, while decreased in ATRA-treated CBDL groups (P<0.05). ATRA (1.5 and 7.5 mg/kg BW) reduced the expression levels of COL Ⅰprotein more greatly than that of 0.1 mg/kg BW (P<0.05). ATRA treatment increased the protein levels of MMP2 and MMP13. The expression levels of TGF-β1 and CTGF mRNA in group A were increased. In comparison with group A, the mRNA levels of TGF-β1 and CTGF in ATRA-treated CBDL groups were significantly decreased (P<0.05). It was concluded that ATRA could inhibit CBDL-induced liver fibrosis in rats by suppressing the expression of TGF-β1 and CTGF so as to diminish the inhibition of TIMP-1 on MMP2 and MMP13 and increase the activity of MMP2 and MMP13.