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ABSTRACT Introduction: The current study about transition of oral epithelial dysplasia, present in lesions such as leukoplakia, for squamous cell carcinoma (SCC) involves not only the histopathological aspects, but also the analysis of the presence of biomarkers which influence the microenvironment where cells are embedded. Objective: To evaluate the tissue inhibitor of metalloproteinase-1 (TIMP-1) profile in cases of leukoplakia and SCC classified into different degrees of dysplasia and histological grading, respectively. The immunohistochemical findings were confronted with microscopic features adopted in the classification of each lesion. Material and methods: Cases of leukoplakia and SCC were recovered from files of The Oral Pathological Anatomy Service of the Dental School at the Universidade Federal do Espírito Santo (SAPB-UFES), between the years 2004 and 2010. New slides were obtained and submitted to immunohistochemical assay to determine TIMP-1 expression profile. Parenchyma, as well as the different layers of the epithelium and stroma was evaluated. Results: In all cases the presence of TIMP-1 was detected in the stroma and parenchyma. In mild leukoplakia, the basal layer with hyperplasia showed intense immunolabeling, whereas cells with loss of polarity presented weaker expression. In moderate leukoplakia, all epithelium layers, except the cornea, were labeled. Severe leukoplakia had the spinous layer most intensely labeled, with no variation in areas with pleomorphism. Stage I SCC showed the deepest islands with intense labeling in cells with pleomorphism and mitoses. In the tumor islands, less differentiated cells were weakly labeled, and in keratin pearl, labeling was weak or absent in central cells. In stage II SCC, labeling was observed in basal cell with hyperplasia and in cells of the spinous layer, however, the parabasal layer was not labeled. Also, on tumor islands, less differentiated cells did not express the protein and keratin pearls were not labeled. Conclusion: It was possible to detect TIMP-1 immunolabeling in all specimens, ranging in intensity and location. The absence of expression in less differentiated cell suggests that more aggressive lesions present reduced enzyme expression. The microenvironment is important for the various cellular activities, and TIMP is an enzyme that participates in matrix remodeling, therefore changes in its expression can be a valuable tool in the better understanding oral carcinogenesis.
RESUMEN Introducción: El estudio actual de la transición de displasia epitelial oral, presente en lesiones como la leucoplasia, hacia carcinoma epidermoide, implica no solo aspectos histopatológicos, sino también el análisis de la presencia de biomarcadores que influyen en el microambiente en el que se insertan las células. Objetivo: Evaluar el perfil del inhibidor tisular metaloproteinasa-1 (TIMP-1) en casos de leucoplasia y carcinoma de células escamosas (CCE) clasificados en diferentes grados de displasia y grados histopatológicos, respectivamente. Confrontar los hallazgos inmunohistoquímicos con los aspectos microscópicos adoptados en la clasificación de lesiones. Material y métodos: Casos de leucoplasia y CCE fueron recuperados del Servicio de Anatomía Patológica Bucal del Curso de Odontología de la Universidad Federal de Espírito Santo (SAPB-UFES), entre los años 2004-2010. Se obtuvieron nuevos portaobjetos y se los sometieron a ensayos inmunohistoquímicos para determinar el perfil de expresión de TIMP-1. Se evaluó el parénquima, así como las diferentes capas del epitelio y estroma. Resultados: En todos los casos se detectó TIMP-1 en estroma y parénquima. En la leucoplasia leve, la capa basal y con hiperplasia mostró inmunotinción intensa, mientras que las células con pérdida de polaridad tuvieron menos expresión. En la leucoplasia moderada, todas las capas del epitelio, excepto la córnea, mostraron inmunotinción. En la leucoplasia grave la capa espinosa tuvo inmunotinción más intensa, sin variación en áreas con pleomorfismo. El CCE grado I mostró las islas más profundas con tinción intensa en células con pleomorfismo y mitosis. En las islas tumorales, las células menos diferenciadas tuvieron tinción menor, y en las perlas de queratina la tinción fue débil o ausente en las células centrales. En el CCE grado II, se observó tinción en células basales con hiperplasia y, en células de la capa espinosa, la capa parabasal no fue marcada. También en las islas, las células menos diferenciadas no expresaron la proteína y no hubo tinción en las perlas de queratina. Conclusión: Fue posible detectar inmunotinción para TIMP-1 en todos los especímenes, con variación en intensidad y ubicación. La ausencia de expresión en células menos diferenciadas sugiere que las lesiones más agresivas tienen enzima reducida. El microambiente es importante para las diversas actividades celulares, y el TIMP es una enzima que participa en la remodelación de la matriz; por lo tanto, la alteración en su expresión puede ser una herramienta valiosa en el mejor entendimiento de la carcinogénesis de la mucosa oral.
RESUMO Introdução: O estudo atual da transição da displasia epitelial oral, presente em lesões como a leucoplasia, para o carcinoma de células escamosas (CCE), envolve não somente aspectos histopatológicos, como também a análise da presença de biomarcadores, os quais influenciam o microambiente em que as células estão inseridas. Objetivo: Avaliar o perfil da expressão do inibidor tecidual de metaloproteinase 1 (TIMP-1) em casos de leucoplasias e CCE classificados em diferentes graus de displasia e graus histopatológicos, respectivamente, e confrontar os achados imuno-histoquímicos com os aspectos microscópicos adotados na classificação das lesões. Material e métodos: Foram resgatados casos de leucoplasia e CCE do Serviço de Anatomia Patológica Bucal do curso de Odontologia da Universidade Federal do Espírito Santo (SAPB-UFES), entre os anos 2004-2010. Novas lâminas foram obtidas ao serem submetidas ao ensaio imuno-histoquímico para determinação do perfil de expressão de TIMP-1. Foram avaliados parênquima, bem como as diferentes camadas do epitélio e estroma. Resultados: Em todos os casos, foi detectada a presença de TIMP-1 no estroma e no parênquima. Na leucoplasia leve, a camada basal e com hiperplasia apresentou imunomarcação intensa; as células com perda de polaridade tiveram expressão menor. Na leucoplasia moderada, todas as camadas do epitélio, exceto a córnea, apresentaram marcação. A leucoplasia severa teve a camada espinhosa marcada mais intensamente, sem variação em áreas com pleomorfismo. O CCE grau I apresentou as ilhas mais profundas com marcação intensa em células com pleomorfismo e mitoses. Nas ilhas tumorais, células menos diferenciadas tiveram marcação menor, e em pérolas córneas a marcação foi fraca ou ausente nas células centrais. No CCE grau II, foi observada a marcação em células basais com hiperplasia e, em células da camada espinhosa, a camada parabasal não foi marcada. Também nas ilhas, células menos diferenciadas não expressaram a proteína e não houve marcação em pérolas córneas. Conclusão: Foi possível detectar imunomarcação para TIMP-1 em todos os espécimes, com variação em intensidade e localização. A ausência de expressão em células menos diferenciadas sugere que lesões mais agressivas possuem redução da enzima. O microambiente é importante para as diversas atividades celulares, e TIMP é uma enzima que participa da remodelação da matriz. Portanto, alteração na sua expressão pode ser uma valiosa ferramenta para um melhor entendimento da carcinogênese da mucosa bucal.
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Objective: To observe the effect of deuterium depleted water combined with platelet-rich plasma on wound healing of diabetic ulcer in rats, and to explore its possible mechanism. Methods: Male SD rats were randomly divided into two groups, normal control group (group A, n=20) and diabetic group (n=80). Rats in the diabetic group were fed with high-fat diet for 4 weeks, and the rat diabetic ulcer model was replicated by intraperitoneal injection of streptozotocin (STZ) + skin full-thickness resection; then randomly divided into diabetic model group (group B), platelet-rich plasma group (group C), deuterium depleted water group (group D), and deuterium depleted water combined platelet-rich plasma group (group E), with 18 rats for each group. Group A with common feed was fed for 4 weeks after intraperitoneal injection of an equal volume of citric acid-sodium citrate buffer + skin full-thickness resection to replicate the normal ulcer model. The animals were sacrificed after treatment for 3, 7 and 14 d, and the random blood glucose was measured at each corresponding time point. The wound surface and wound margin tissue were taken to observe the wound healing and local histomorphology of each group. The contents of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the wound tissue of each group were detected by enzyme-linked reaction adsorption method. Results: Random blood glucose in group D and group E was lower than that before intervention. The inflammatory response of the wounds in each diabetic group was slower than that in group A. The granulation ripening effect of group E was faster than that of group B, C, and D. The effect was best in each intervention group, and the neovascularization and fibroblasts appeared earlier and in large quantities. The content of TIMP-1 in group A was significantly higher than that in group B, C, D and E (P<0.05). The content of TIMP-1 in group B was significantly lower than that in group C, D and E (P<0.05). The content of TIMP-1 was significantly higher than that of group C and D (P<0.05). The content of MMP-9 in group B was significantly higher than that in group A, C, D and E (P<0.05). The content of MMP-9 in group E was significantly lower than that in group C and D (P<0.05). Conclusion: Deuterium depleted water can promote the healing of diabetic ulcer wounds. Deuterium depleted water combined with platelet-rich plasma can significantly promote the healing of diabetic ulcer wounds, which may be related to the decrease of random blood glucose, the increase of TIMP-1 and the inhibition of MMP-9 expression.
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BACKGROUND: In the development of osteoarthritis, the mechanism underlying cartilage damage is still unclear. Matrix metalloproteinases have been shown to play important roles in cartilage matric degradation. OBJECTIVE: To observe the correlation between the expression of matrix metalloproteinase 3, tissue inhibitor of metalloproteinase-1 and the pathological degree in knee osteoarthritis. METHODS: The study was approved by the Ethics Committee of Shenyang Orthopedic Hospital. The tibial plateaus of 40 patients with knee osteoarthritis who underwent total knee arthroplasty were collected, and all patients signed the informed consents. The classification of osteoarthritis was evaluated according to Kellgren-Lawrence (KL) scale system: 11 cases of KL grade 2, 15 cases of KL grade 3, and 14 cases of KL grade 4. Control group contained six cases. Samples received hematoxylin-eosin staining. The pathological changes of knee osteoarthritis cartilage were evaluated by Mankin score. Immunohistochemical staining was used to detect the contents of matrix metalloproteinase 3 and tissue inhibitor of metalloproteinase-1 in cartilage. RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining results showed that in the control group, the layer of cartilage was thick, and there were abundant chondrocytes that arranged regularly. The subcutaneous layer of cartilage in the KL grade 2 group was rugged, and fissure was observed occasionally. Fibrosis of cartilage layer was visible in the KL grade 3 group, and the chondrocytes arranged in disorder. In the KL grade 4 group, the structure of cartilage layer was lost, and there were few chondrocytes that arranged irregularly. (2) Immunohistochemical staining results showed that the expression level of matrix metalloproteinase 3 in the osteoarthritis group was significantly higher than that in the control group. The expression level of matrix metalloproteinase 3 was on a rise in the KL grade 2, 3 and 4 groups (all P < 0.05). The expression level of tissue inhibitor of metalloproteinase-1 in the osteoarthritis group was significantly lower than that in the control group. The expression level of tissue inhibitor of metalloproteinase-1 was on a descent in the KL grade 2, 3 and 4 groups (all P < 0.05). (3) There was significantly positive correlation between matrix metalloproteinase 3 expression level and Mankin score (r=0.899, P < 0.001), and tissue inhibitor of metalloproteinase-1 expression level was negatively correlated with Mankin score (r=-0.903, P < 0.001). There was significantly negative correlation between tissue inhibitor of metalloproteinase-1 expression level and matrix metalloproteinase 3 expression level (f=-0.881, P < 0.001). (4) These results indicate that in the articular cartilage of patients with osteoarthritis, the expression levels of matrix metalloproteinase 3 and tissue inhibitor of metalloproteinase-1 are correlated with the pathological changes, which can be used as an effective index to evaluate the progress of knee osteoarthritis.
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BACKGROUND: Liver fibrosis has higher morbidity and mortality. Activation and proliferation of hepatic stellate cells is a key link in the progression of liver fibrosis. At present, there are still no effective anti-fibrosis agents targeting single links or targets. OBJECTIVE: To analyze the effect of human adipose stem cells derived exosomes on rats with liver fibrosis induced by carbon tetrachloride. METHODS: Human adipose stem cells were obtained from healthy people by enzyme dissolution method. After in vitro culture, human adipose stem cells derived exosomes were obtained by multiple ultrafiltration. Different concentrations of exosomes were used to treat the hepatic stellate cells activated by transforming growth factor β1. The human adipose stem cells activated by transforming growth factor β1 were treated with different concentrations of exosomes. The expression of α-smooth actin in the cells was detected by quantitative PCR, and the growth and apoptosis of activated hepatic stellate cells were detected by CCK-8 and flow cytometry respectively. Rat models of liver fibrosis were established by intraperitoneal injection of carbon tetrachloride and treated by tail vein injection of exosomes. Rat liver function, serum levels of type III procollagen and type IV collagen, and Ishak score were determined. Semi-quantitative analysis of liver fibrosis was performed. The expression levels of tissue inhibitor of matrix metalloproteinase-1, matrix metalloproteinase 9 and α-smooth actin in liver tissue were measured by immunofluorescence method. The study protocol was approved by the Animal Ethics Committee and Medical Ethics Committee, Tongji University, China in January, 2017. RESULTS AND CONCLUSION: Human adipose stem cells derived exosomes inhibited the proliferation of activated hepatic stellate cells. The possible mechanism is to inhibit the proliferation of activated macrophages, reduce the production of collagen fibers, α-smooth actin actin, and tissue inhibitor of matrix metalloproteinase-1, and to increase the expression of matrix metalloproteinase 9. These findings suggest that exosomes can be used to treat carbon tetrachloride induced liver fibrosis.
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OBJECTIVE@#To investigate the effect of Modified Xiaochaihu Decoction (MXD, ) on collagen degradation in rats with chronic pancreatitis (CP).@*METHODS@#Rats were injected dibutyltin dichloride (DBTC, 7 mg/kg of body weight) into the right caudal vein to induce CP model. Thirty heallhy male Wistar rats were randomly divided into three groups by a random number table: the control, the model and the treatment groups. Rats of treatment group were administered MXD (10 g/kg of body weight) orally once daily starting from the day post-model establishment. Pancreatic tissues were harvested after 28-day feeding and fibrosis was evaluated by picro-sirius red staining. The contents of collagen type I and III were detected using enzymelinked immunosorbent assay (ELISA), the expression of matrix metalloproteinase 13 (MMP13) and tissue inhibitor of metalloproteinase 1 (TIMP1) was analyzed by Western blot and real-time polymerase chain reaction (PCR).@*RESULTS@#The fibrosis scoring of pancreatic tissues, the concentrations of collagen type I and III, the expression levels of MMP13 and TIMP1 proteins and mRNA in the model group were all increased compared with the control group (P0.05).@*CONCLUSIONS@#MXD could promote collagen degradation and reverse pancreatic fibrosis in CP rats via a mechanism involve up-regulation of MMP13 expression.
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BACKGROUND: Liver fibrosis has higher morbidity and mortality. Activation and proliferation of hepatic stellate cells is a key link in the progression of liver fibrosis. At present, there are still no effective anti-fibrosis agents targeting single links or targets.OBJECTIVE: To analyze the effect of human adipose stem cells derived exosomes on rats with liver fibrosis induced by carbon tetrachloride. METHODS: Human adipose stem cells were obtained from healthy people by enzyme dissolution method. After in vitro culture, human adipose stem cells derived exosomes were obtained by multiple ultrafiltration. Different concentrations of exosomes were used to treat the hepatic stellate cells activated by transforming growth factor β1. The human adipose stem cells activated by transforming growth factor β1 were treated with different concentrations of exosomes. The expression of α-smooth actin in the cells was detected by quantitative PCR, and the growth and apoptosis of activated hepatic stellate cells were detected by CCK-8 and flow cytometry respectively. Rat models of liver fibrosis were established by intraperitoneal injection of carbon tetrachloride and treated by tail vein injection of exosomes. Rat liver function, serum levels of type III procollagen and type IV collagen, and Ishak score were determined. Semi-quantitative analysis of liver fibrosis was performed. The expression levels of tissue inhibitor of matrix metalloproteinase-1, matrix metalloproteinase 9 and α-smooth actin in liver tissue were measured by immunofluorescence method. The study protocol was approved by the Animal Ethics Committee and Medical Ethics Committee, Tongji University, China in January, 2017. RESULTS AND CONCLUSION: Human adipose stem cells derived exosomes inhibited the proliferation of activated hepatic stellate cells. The possible mechanism is to inhibit the proliferation of activated macrophages, reduce the production of collagen fibers, α-smooth actin actin, and tissue inhibitor of matrix metalloproteinase-1, and to increase the expression of matrix metalloproteinase 9. These findings suggest that exosomes can be used to treat carbon tetrachloride induced liver fibrosis.
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AIM:To investigate the effects of Maxing-Shigan decoction on airway remodeling and expression of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the lung tissues of asthmatic mice, and to explore its possible mechanism in treatment of asthma.METHODS:The BALB/c mice were divided into blank control group, model group, low-dose Maxing-Shigan decoction group, middle-dose Maxing-Shigan decoction group, high-dose Maxing-Shigan decoction group and positive control group.The mice were sensitized and challenged with ovalbumin to establish asthma model.The mice in blank control group and model group were given saline by oral administration before 30 min of suscitation.The mice in low-dose, middle-dose and high-dose Maxing-Shigan decoction groups were given Maxing-Shigan decoction at 5.0 g/kg, 10.0 g/kg and 20.0 g/kg, respectively, by oral administration before 30 min of suscitation.The mice in positive control group was given dexamethasone at 0.005 g/kg by oral administration before 30 min of suscitation.After consecutive administration for 7 d, the variations of airway responsiveness, the percentage of the goblet cells, the collagen deposition, and the eosinophil (EOS) counts in bronchoalveolar lavage fluid (BALF) of each group were observed.The protein levels of MMP-9 and TIMP-1 in the lung tissues were determined by ELISA and Western blot.The mRNA expression of MMP-9 and TIMP-1 was detected by RT-qPCR.RESULTS:Compared with blank control group, the airway responsiveness, the goblet cell percentage, the collagen deposition, the EOS counts in BALF, the protein levels of MMP-9 and TIMP-1, and the mRNA expression of MMP-9 and TIMP-1 were significantly increased in model group (P<0.01).Compared with model group, all of the indexes were reversed in low-dose, middle-dose and high-dose Maxing-Shigan decoction groups and positive control group (P<0.05 or P<0.01).CONCLUSION:Maxing-Shigan decoction improves airway remodeling in asthma model mice by down-regulating the expression of MMP-9 and TIMP-1.
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Objective To investigate the effect of anisodamine combined with blood purification on pulmonary fibrosis and levels of serum matrix metalloproteinase-9 (MMP-9) and matrix metalloproteinase 1 (TIMP1) in patients with paraquat poisoning.Methods From March 2013 to February 2017,84 patients with paraquat poisoning admitted to our hospital were enrolled in the observation group (anisodamine +blood purification) or control group (blood purification),42 cases in each group.The curative effect,indexes related to pulmonary fibrosis,serum MMP-9 and TIMP1 levels were compared between the two groups.The incidence of acute lung injury/acute respiratory distress syndrome (ALI/ARDS) and intensive care uni (ICU) admission time were recorded.Results There was significant difference (P ＜ 0.05) in the total effective rate (88.1％ vs 69.0％) and the high-resolution computed tomography (CT) score after 7 days of treatment (8.4 ± 0.9 vs 12.3 ± 1.4) between the observation group and the control group.At the same time,the serum levels of hyaluronic acid and type Ⅳ collagen in the two groups showed an upward trend with significant difference (P ＜ 0.05).After 1 day of treatment,the serum levels of MMP-9 and TIMP1 in the observation group reached a peak,then gradually decreased,with significant difference (P ＜0.05).The incidence of ALI/ARDS in the observation group and the control group (11.9％ vs 31.0％)and the duration of ICU treatment [(10.2 ± 3.2) d vs (13.2 ± 2.8) d].Conclusions For patients with acute paraquat poisoning,the combination of anisodamine and blood purification can down-regulate the expressions of MMP-9 and TIMP1 and delay the progression of pulmonary fibrosis,thus improving the clinical therapeutic effect.
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Objective@#To investigate the effect of three-dimensional conformal radiotherapy (3D-CRT) combined with PC chemotherapy (paclitaxel + carboplatin) on non-small cell lung cancer (NSCLC) patients and the serum levels of CA125, tissue inhibitor of metalloproteinase-1 (TIMP-1), serum amyloid A (SAA) and T-lymphocyte subsets.@*Methods@#A total of 100 patients with NSCLC treated in Affiliated Hospital of Guangdong Medical University from May 2015 to December 2017 were selected as the study subjects. They were divided into control group and observation group according to random number table method, with 50 cases in each group. The observation group was treated with 3D-CRT combined with PC chemotherapy, while the control group was treated with PC chemotherapy. The two groups were treated for 4 cycles. The therapeutic effect, serum CA125, TIMP-1, SAA, T-lymphocyte subsets and adverse reactions were compared between the two groups.@*Results@#Four cases were lost to follow-up both in the two groups. The overall response rate in the observation group (43.48%, 20/46) was higher than that in the control group (23.91%, 11/46; χ2=3.941, P=0.047). The serum levels of CA125, TIMP-1 and SAA of the two groups had no significant difference before treatment, and the levels of these indexes decreased after treatment. The serum levels of CA125, TIMP-1 and SAA in the observation group after treatment were (12.31±1.13) U/ml, (275.31±13.69) pg/ml and (47.21±7.21) mg/L, which were lower than those in the control group [(30.36±1.98) U/ml, (320.36±17.23) pg/ml, (65.92±8.36) mg/L], with significant differences (t=53.699, P<0.001; t=13.884, P<0.001; t=11.495, P<0.001). The levels of CD3+ , CD4+ , CD8+ and CD4+ /CD8+ of the two groups had no significant difference before treatment, and the levels of these indexes decreased after treatment. The levels of CD3+ , CD4+ , CD8+ and CD4+ /CD8+ in the observation group were (35.27±10.31)%, (20.27±6.72)%, (15.89±3.37)% and 0.91±0.37, which were higher than those in the control group [(30.77±9.27)%, (15.27±5.73)%, (12.02±2.69)% and 0.75±0.39], with significant differences (t=2.201, P=0.030; t=3.840, P<0.001; t=6.087, P<0.001; t=2.019, P=0.047). There were no significant differences in the adverse reactions such as nausea and vomiting [63.04% (29/46) vs. 43.48% (20/46); χ2=3.537, P=0.060], phlebitis [6.52% (3/46) vs. 4.35% (2/46); χ2=0.000, P>0.999], abnormal liver function [6.52% (3/46) vs. 2.17% (1/46); χ2=0.261, P=0.609] and myelosuppression [8.70% (4/46) vs. 6.52% (3/46); χ2=0.000, P>0.999] between the observation group and the control group.@*Conclusion@#For patients with NSCLC, 3D-CRT combined with PC chemotherapy can improve the overall response rate, decrease the levels of serum CA125, TIMP-1 and SAA, and improve the immune function of patients. The therapeutic effect is remarkable and the safety is good. The therapeutic scheme is suitable for the treatment of NSCLC.
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Objective@#To investigate the effect of anisodamine combined with blood purification on pulmonary fibrosis and levels of serum matrix metalloproteinase-9 (MMP-9) and matrix metalloproteinase 1 (TIMP1) in patients with paraquat poisoning.@*Methods@#From March 2013 to February 2017, 84 patients with paraquat poisoning admitted to our hospital were enrolled in the observation group (anisodamine + blood purification) or control group (blood purification ), 42 cases in each group. The curative effect, indexes related to pulmonary fibrosis, serum MMP-9 and TIMP1 levels were compared between the two groups. The incidence of acute lung injury/acute respiratory distress syndrome (ALI/ARDS) and intensive care uni (ICU) admission time were recorded.@*Results@#There was significant difference (P<0.05) in the total effective rate (88.1% vs 69.0%) and the high-resolution computed tomography (CT) score after 7 days of treatment (8.4±0.9 vs 12.3±1.4) between the observation group and the control group. At the same time, the serum levels of hyaluronic acid and type Ⅳ collagen in the two groups showed an upward trend with significant difference (P<0.05). After 1 day of treatment, the serum levels of MMP-9 and TIMP1 in the observation group reached a peak, then gradually decreased, with significant difference (P<0.05). The incidence of ALI/ARDS in the observation group and the control group (11.9% vs 31.0%) and the duration of ICU treatment [(10.2±3.2)d vs (13.2±2.8)d].@*Conclusions@#For patients with acute paraquat poisoning, the combination of anisodamine and blood purification can down-regulate the expressions of MMP-9 and TIMP1 and delay the progression of pulmonary fibrosis, thus improving the clinical therapeutic effect.
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Objective@#To explore the expression levels of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) protein and the change of MMP-9/TIMP-1 ratio in wound exudates of patients with stages Ⅲ and Ⅳ pressure ulcers during wound healing.@*Methods@#From July 2017 to July 2018, 30 patients with stage Ⅲ pressure ulcers [30 wounds, 16 males and 14 females, aged (65±10) years] and 34 patients with stage Ⅳ pressure ulcers [50 wounds, 17 males and 17 females, aged (65±9) years] admitted to Hebei General Hospital who met the inclusion criteria were enrolled in this prospective cohort study. According to the principle of wound treatment and the characteristics and needs of wound in different periods, individualized intervention measures were formulated for patients and appropriate dressings were selected. At the time of admission and on 7, 14, 21, 28 days of treatment, the healing of pressure ulcer wounds was evaluated by Pressure Ulcer Healing Scale. Afterwards, the wound exudate was collected at each time point to detect the expression levels of MMP-9 and TIMP-1 protein by enzyme-linked immunosorbent assay, and the MMP-9/TIMP-1 ratio was calculated. Data were processed with analysis of variance for repeated measurements of single group and linear trend test.@*Results@#(1) There were significantly statistical differences in wound healing scores of patients with stages Ⅲ and Ⅳ pressure ulcers among the time of admission and on 7, 14, 21, 28 days of treatment within each stage (F=145.382, 153.234, P<0.01), and they all showed a gradually decreasing trend (F=170.466, 284.585, P<0.01). (2) At the time of admission and on 7, 14, 21, 28 days of treatment, the expression levels of MMP-9 protein in wound exudates of patients with stages Ⅲ and Ⅳ pressure ulcers were (171±104), (138±88), (110±70), (85±55), (62±41) ng/L and (193±107), (173±104), (139±83), (114±70), (89±56) ng/L, respectively. There were significantly statistical differences within each stage (F=58.007, 111.680, P<0.01), and they all showed a gradually decreasing trend (F=62.901, 134.628, P<0.01). At the time of admission and on 7, 14, 21, 28 days of treatment, the expression levels of TIMP-1 protein in wound exudates of patients with stages Ⅲ and Ⅳ pressure ulcers were (6.2±3.9), (5.6±3.4), (5.1±3.1), (4.4±2.5), (3.8±2.3) ng/L and (4.8±2.5), (4.7±2.6), (4.4±2.6), (4.6±2.7), (4.1±2.4) ng/L, respectively. There were significantly statistical differences within each stage (F=25.479, 7.778, P<0.01), and there was a gradually decreasing trend in stage Ⅲ (F=62.901, P<0.01) and a decreasing trend in stage Ⅳ (F=134.628, P<0.01). At the time of admission, the expression levels of MMP-9 and TIMP-1 in wound exudates of patients with stage Ⅲ pressure ulcers were similar to those of patients with stage Ⅳ pressure ulcers (t=-1.03, 1.47, P>0.05). (3) At the time of admission and on 7, 14, 21, 28 days of treatment, the MMP-9/TIMP-1 ratios in the wound exudates of patients with pressure ulcers of stages Ⅲ and Ⅳ were 30±13, 25±9, 22±9, 20±8, 17±6 and 43±19, 37±13, 32±10, 26±9, 22±9, respectively. There were significantly statistical differences within each stage (F=37.173, 97.191, P<0.01), and they all showed a gradually decreasing trend (F=54.183, 130.088, P<0.01). At the time of admission, the MMP-9/TIMP-1 ratio in wound exudates of patients with stage Ⅳ pressure ulcers was significantly higher than that of patients with stage Ⅲ pressure ulcers (t=-3.42, P<0.01).@*Conclusions@#During the wound healing process of patients with stages Ⅲ and Ⅳ pressure ulcers, the expression levels of MMP-9 and TIMP-1 protein and the MMP-9/TIMP-1 ratio in wound exudates show a decreasing trend. The stage of wound healing can be predicted according to the expression level of MMP-9 protein and the MMP-9/TIMP-1 ratio.
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OBJECTIVE： To study improvement effects of Panax notoginsenoside（PNS） on cisplatin-induced renal injury model rats and its effects on related factors. METHODS： Totally 72 SD rats were randomly divided into blank group， model group， positive drug group and PNS low-dose， medium-dose， high-dose groups， with 12 rats in each group. Except for blank group， other groups were given cisplatin via tail vein （3 mg/kg×4 times） to establish renal injury model. Since the first day after the first injection of cisplatin， positive group was given anfostine solution intraperitoneally （1.0 mg/kg）； PNS groups were given PNS solution intraperitoneally （15.63， 31.35， 62.70 mg/kg）； blank group and model groups were given constant volume of normal saline 0.2 mL， for consecutive 60 d. The 24 h urine of rats was collected； the contents of β-N-acetylaminoglycosidase（NAG） and 24 h urine protein （Upro/24 h） were detected； the serum contents of Scr and BUN were detected. mRNA and protein expression of CTGF， TGF-β1， Col-1， TIMP-1 and PAI-1 in renal tissue were determined by RT-PCR and immunohistochemistry respectively. RESULTS： Compared with blank group， the contents of NAG and Upro/24 h in urine， serum contents of Scr and BUN， mRNA and protein expression levels of CTGF， TGF-β1， Col-1， TIMP-1 and PAI-1 in renal tissue were increased significantly （P＜0.05）. Compared with model group， the contents of above urine and serum biochemical indicators were decreased significantly in PNS groups； mRNA expression of CTGF and TGF-β1 and protein expression of CTGF， TGF-β1， Col-1 and TIMP-1 in renal tissue of rats in PNS groups， mRNA expression of Col-1 in PNS high-dose group， and mRNA expression of TIMP-1 and protein expression of PAI-1 in PNS medium-dose and high-dose groups were decreased significantly （P＜0.05）. Compared with positive group， the contents of NAG and Upro/24 h in urine were decreased significantly in PNS medium-dose and high-dose groups （P＜0.05）. CONCLUSIONS： PNS can effectively improve the renal function of cisplatin-induced renal injury model rats， and relieve cisplatin-induced renal fibrosis by decreasing the expression of renal fibrosis related factors as CTGF， TGF-β1， Col-1， TIMP-1 and PAI-1 in renal tissue.
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Objective To investigate the effect of three-dimensional conformal radiotherapy (3D-CRT)combined with PC chemotherapy (paclitaxel + carboplatin)on non-small cell lung cancer (NSCLC) patients and the serum levels of CA125,tissue inhibitor of metalloproteinase-1 (TIMP-1),serum amyloid A (SAA)and T-lymphocyte subsets. Methods A total of 100 patients with NSCLC treated in Affiliated Hospital of Guangdong Medical University from May 2015 to December 2017 were selected as the study subjects. They were divided into control group and observation group according to random number table method,with 50 cases in each group. The observation group was treated with 3D-CRT combined with PC chemotherapy,while the con-trol group was treated with PC chemotherapy. The two groups were treated for 4 cycles. The therapeutic effect, serum CA125,TIMP-1,SAA,T-lymphocyte subsets and adverse reactions were compared between the two groups. Results Four cases were lost to follow-up both in the two groups. The overall response rate in the observation group (43. 48％,20 / 46)was higher than that in the control group (23. 91％,11 / 46;χ2 = 3. 941, P = 0. 047). The serum levels of CA125,TIMP-1 and SAA of the two groups had no significant difference be-fore treatment,and the levels of these indexes decreased after treatment. The serum levels of CA125,TIMP-1 and SAA in the observation group after treatment were (12. 31 ± 1. 13)U/ ml,(275. 31 ± 13. 69)pg/ ml and (47. 21 ± 7. 21)mg/ L,which were lower than those in the control group [(30. 36 ± 1. 98)U/ ml,(320. 36 ± 17. 23)pg/ ml,(65. 92 ± 8. 36)mg/ L],with significant differences (t = 53. 699,P < 0. 001;t = 13. 884, P < 0. 001;t = 11. 495,P < 0. 001). The levels of CD3 +,CD4 +,CD8 + and CD4 + / CD8 + of the two groups had no significant difference before treatment,and the levels of these indexes decreased after treatment. The levels of CD3 +,CD4 +,CD8 + and CD4 + / CD8 + in the observation group were (35. 27 ± 10. 31 )％, (20. 27 ± 6. 72)％,(15. 89 ± 3. 37)％ and 0. 91 ± 0. 37,which were higher than those in the control group [(30. 77 ± 9. 27)％,(15. 27 ± 5. 73)％,(12. 02 ± 2. 69)％ and 0. 75 ± 0. 39],with significant differences (t = 2. 201,P = 0. 030;t = 3. 840,P < 0. 001;t = 6. 087,P < 0. 001;t = 2. 019,P = 0. 047). There were no significant differences in the adverse reactions such as nausea and vomiting [63. 04％ (29 / 46)vs. 43. 48％(20 / 46);χ2 = 3. 537,P = 0. 060],phlebitis [6. 52％ (3 / 46)vs. 4. 35％ (2 / 46);χ2 = 0. 000,P >0. 999],abnormal liver function [6. 52％ (3 / 46)vs. 2. 17％ (1 / 46);χ2 = 0. 261,P = 0. 609]and myelo-suppression [8. 70％ (4 / 46)vs. 6. 52％ (3 / 46);χ2 = 0. 000,P > 0. 999]between the observation group and the control group. Conclusion For patients with NSCLC,3D-CRT combined with PC chemotherapy can im-prove the overall response rate,decrease the levels of serum CA125,TIMP-1 and SAA,and improve the im-mune function of patients. The therapeutic effect is remarkable and the safety is good. The therapeutic scheme is suitable for the treatment of NSCLC.
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Objective To explore the expressions and clinical significance of matrix metalloproteinase9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in renal carcinoma.Methods A total of 45 patients with renal carcinoma received treatment at our hospital from January 2015 to January 2017 were enrolled as observation group,and 40 healthy subjects in the same period were enrolled as control group.The expressions of MMP-9 and TIMP-1 in observation group and control group were tested and compared by enzymelinked immunosorbent assay.The relationship between MMP-9 and TIMP-1 and their correlations with the clinical pathological features were analyzed.Results The serum levels of MMP-9 in observation group were higher than those in control group,and the differences were significantly significant [(461.7 ± 53.6) pg/ml vs.(142.9±31.7) pg/ml;t=32.840,P＜0.001],TIMP-1 [(387.9±11.8) pg/ml vs.(136.5 ±21.8) pg/ml;t=13.290,P＜0.001] andMMP-9/TIMP-1 [(1.2±0.2) vs.(1.1 ±0.1);t=3.640,P＜0.001].Correlation analysis showed that the serum expression level of MMP-9 had significantly positive correlation with TIMP-1 for patients in observation group (r =0.847,P ＜ 0.001).The serum expression levels of MMP-9 and TIMP-1 for patients in observation group were related to the lymph node state (t =2.657,P =0.011;t =4.420,P ＜0.001) and primary tumor staging (t =6.200,P ＜0.001;t =5.042,P ＜0.001),but they were not related to pathological type (t =1.130,P =0.265;t =0.109,P =0.914).Conclusion The peripheral blood levels of MMP-9 and TIMP-1 are increased in patients with renal carcinoma,and they are associated with the progression of disease,which can be used to evaluate the progress of disease.
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Objective To study on the relationship between the serum level of highsensitivity c-reaction protein (hs-CR),matrix metalloproteinase 9 (MMT-9),tissue inhibitor of metalloproteinase 1 (TIMP-1) and carotid plaque stability in elderly people.Methods According to the carotid ultrasonography examination,120 old people were divided into vulnerable plaque group (group A,n =45),stable plaque group (group B,n =41) and no plaque group (group C,n =34),and serum levels of hs-CRP,MMP-9,TIMP-1 in each group were also detected.Results The serum levels of hs-CRP and MMP-9 of unstable plaque group and stable plaque group were higher than those of non-plaque group (P ＜ 0.05).hsCRP and MMP-9 were positively correlated with carotid plaque unstability,while TIMP-1 was negatively correlated with carotid plaque unstability,hs-CRP,MMP-9 and TIMP-1 were independent risk factors for carotid plaque stability.Conclusions Serum levels of hs-CRP,MMP-9 and TIMP-1 are closely related to the stability of carotid plaque.Elevated levels of hs-CRP and MMP-9 increase the risk of carotid plaque,and elevated levels of TIMP-1 decrease the risk of carotid plaque.
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AIM:To determine the expression of matrix metalloproteinase-9(MMP-9),tissue inhibitor of me-talloproteinase-1(TIMP-1)and collagen type Ⅳ(Ⅳ-C)in the lung of rats with multiple organ dysfunction syndrome(MODS)and to investigate the mechanism of lung injury in MODS.METHODS:Adult male Sprague-Dawley(SD)rats(n=40)were randomly divided into sham control group and cecal ligation and puncture(CLP)model group.The rats in CLP group were divided into 4 subgroups as different intervals(6 h,12 h,24 h and 48 h),and there were 8 rats in each group.The rat model of MODS was established by CLP.All rats were sacrificed at various intervals.The functions of the liver,kidney and lung were determined by blood biochemical and blood gas analysis.The morphological changes of the lung tissues were observed with HE staining.The serum levels of tumor necrosis factor(TNF)-α,interleukin(IL)-1β,MMP-9 and TIMP-1 were measured by ELISA.The expression of MMP-9 and TIMP-1 in the lung tissues was detected by RT-PCR and immunohistochemistry ,and the expression of Ⅳ-C in the lung tissues was detected by immunofluorescence and Western blot.RESULTS:Compared with sham control group ,the functions of the liver ,kidney and lung were damaged at different degrees in model groups.No histopathological change in the lung tissues of sham control group was found ,and the lung injury was serious in model groups.Compared with sham control group ,the serum levels of TNF-α,IL-1β,MMP-9 and TIMP-1 in model groups increased significantly(P<0.05)and peaked at the interval of 12 ～24 h after modeling(P<0.01).The expression of MMP-9 and TIMP-1 in the lung tissues of model groups increased ,and peaked at 12 and 24 h,respectively(P<0.01).The protein level of Ⅳ-C in MODS 6 h group was not changed as compared with control group,while that at the interval of 12～48 h after modeling was significantly decreased and dropped to the lowest at 24 h(P<0.01).CONCLUSION:MMP-9 and TIMP-1 play important roles in lung injury of MODS rats by regulating the syn-thesis and decomposition of IV-C which is the main component of extracellular matrix.
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Objective To study the clinical value of 16-slice spiral CT combined with MMP-9 and TIMP-1 in assessing carotid atherosclerosis in elderly cerebral infarction(CI) patients.Methods One hundred and eighty-six CI patients served as group A and 190 asymptomatic carotid atherosclerosis patients served as group B with 90 healthy subjects served as a control group in this study.The patients in group A were further divided into unstable plaque(A-1) group,stable plaque(A-2) group and plaque-free(A-3) group while those in group B were further divided into unstable plaque(B-1) group,stable plaque(B-2) group and plaque-free(B-3) group.The patients included in this study underwent multislice spiral CT angiography(MSCTA) and Doppler ultrasonography.Their serum levels of MMP-9 and TIMP-1 were measured.Their carotid artery steno sis and plaques were detected.The efficacy of MMP-9 and TIMP-1 in diagnosis of unstable plaques in CI patients was analyzed.Results The rate of mild stenosis detected by MSCTA was significantly lower while that of severe stenosis detected by MSCTA was significantly higher in group A than in group B(23.1％ vs 48.9％,P=0.000;48.4％ vs 16.8％,P=0.000).The rate of unstable plaques detected by MSCTA and ultrasonography was 81.7％ and 83.2％ respectively in groups A and B.The rate of stable and unstable plaques detected by MSCTA was significantly higher in group A than that detected by CT in group B(147±71 HU vs 11±20 HU,155±81 HU vs 9±21 HU,P=0.000).The serum levels of MMP-9 and TIMP-1 were significantly higher in A-1 group and A-2 group,B-1 group and B-2 group than in control group(P＜0.01).The area under the ROC curve for MMP-9 and TIMP-1 in diagnosis of unstable plaques was 0.891 and 0.814 respectively.Conclusion MSCTA can accurately detect carotid atherosclerosis in elderly CI patients.However,it is easy to misjudge the nature of plaques.Serum levels of MMP-9 and TIMP-1 can effectively diagnose unstable plaques and can thus be used as an important supplement to MSCTA.
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Objective@#To observe the effects of recombinant adenovirus with human tissue inhibitor of metalloproteinase-1(Ad-hTIMP-1) on the inflammatory response in rats with myocardial infarction (MI) and explore the related mechanisms.@*Methods@#The male Wistar rats were randomly divided into sham-operated group, saline group, Ad-Track group and Ad-hTIMP-1 group according to the random number table (n=8 each group). MI was induced by ligation of the left anterior descending coronary artery and MI rats were injected with saline, Ad-Track and Ad-hTIMP-1, respectively. Sham-operated rats received similar surgical procedure without ligation of the left anterior descending coronary artery. After 4 weeks, the cardiac function was measured by echocardiography, then rats were sacrificed and hearts were removed for morphological and biological analysis. The morphology of myocardial tissue in each group was detected by HE staining and Masson staining. The mRNA expressions of tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-10 and C-reactive protein(CRP) were detected by real-time PCR. Immune histochemical staining was performed to observe the protein expression levels of IL-6 and CRP.@*Results@#(1) Left ventricular end systolic dimension derived from echocardiography was increased in saline group ((5.10±0.72) mm) and Ad-Track group ((4.88±0.64) mm) compared to sham-operated group ((4.25±0.46) mm), which was reduced in Ad-hTIMP-1 group ((4.13±0.35) mm, all P<0.05). The left ventricular ejection fraction was (72.46±5.74)%, (64.27±8.52)%, (64.65±3.90)%, and (71.55±6.95)%, the fractional shortening was (36.90±4.97)%, (29.03±3.40)%, (30.95±2.51)%, and (36.31±5.68)% in sham-operated group, saline group, Ad-Track group and Ad-hTIMP-1 group, respectively. The left ventricular ejection fraction and fractional shortening in saline group and Ad-Track group were lower than those in sham-operated group and Ad-hTIMP-1 group (all P<0.05). (2) Necrosis of myocardial cells was not found and a small amount of immune cell infiltration and interstitial fibrosis were observed on HE and Masson stained myocardial sections of Ad-hTIMP-1 group. (3) Real-time PCR showed that mRNA expressions of TNF-α, IL-6, IL-10 and CRP were lower in Ad-hTIMP-1 group than in saline group. mRNA expressions of TNF-α, IL-10 and CRP were lower in Ad-hTIMP-1 group than in Ad-Track group (all P<0.05). (4) Immune histochemical staining showed that protein expressions of IL-6 and CRP were higher in saline group and Ad-Track group than those in Ad-hTIMP-1 group (all P<0.05).@*Conclusion@#Recombinant adenovirus Ad-hTIMP-1 can improve cardiac function in rats with myocardial infarction via inhibiting the inflammatory response and downregulating the expression of TNF-α, IL-6 and CRP.
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Objective To investigate the expressions of metalloproteinase-9(MMP-9)in serum,brain and aorta matrix and tissue inhibitor of metalloproteinase-1(TIMP-1) in renovascular hypertensive rats(RHR),and to evaluate the association between blood pressure and levels of matrix metalloproteinase-9(MMP-9) and tissue inhibitor of metalloproteinase-1(TIMP-1).Methods Eighty healthy male SD rats were randomly divided into RHR group(n=40) and sham-operated group(n=40).Hypertension was induced by two-kidney,two-clip(2K-2C)clamps.Systolic blood pressure(SBP) was measured every 2 weeks during 12 weeks using a tail pressure meter.Stroke was confirmed by Longa's five-point scale and pathological examination.The expressions of MMP-9 and TIMP-1 in the brain and aorta tissues were detected by Western blot and immunohistochemistry.The levels of serum MMP-9 and TIMP-1 were measured by double-antibody sandwich enzyme-linked immunosorbent assay(DAS-ELISA).Results Compared with the sham-operated group,SBP stayed significantly elevated in the RHR group at 2,4,6,8,10 and 12weeks after the operation [(157±9.0) vs.(128±7.0),(176±10.0) vs.(122±6.0),(194±8.0)vs.(117±6.5),(202±12.0)vs.(124±8.0),(218±15.0) vs.(126.±8.5),and(224±20.0)vs.(129.±9.0) mm Hg,all P< 0.05].12 weeks after the surgery,the level of serum MMP-9 in the RHR group was kept significantly higher than that in the sham-operated group [(783.4±109.79)vs.(573.4±109.59) ng/mL,P<0.05],and the serum TIMP-1 level was lower in the RHR group than that in the sham-operated group[(313.02±83.9) vs.(976.19±191.1) pg/mL,P<0.05].MMP-9 expressions were significantly higher in the brain and aorta in the RHR group than that in the sham-operated group(both P<0.05),and TIMP-1 expressions were lower than that in the sham-operated group(both P<0.05).The Pearson correlation analysis showed that MMP-9 levels in serum,brain and aorta were positively correlated with systolic blood pressure(r=0.557,r=0.774 and r=0.661,all P<0.05),and TIMP-1 levels were negatively correlated with systolic blood pressure(r=-0.481,r=-0.535 and r=-0.685,all P<0.01).Conclusions Hypertension induces increased MMP-9 and decreased TIMP-1 in serum,brain and aorta in renovascular hypertensive rats.There are consistent alterations of circulating and tissue MMP-9 and TIMP-1 levels in renovascular hypertensive rats.There is a relationship between increased blood pressure and high MMP-9 and low TIMP-1 in serum and tissues.
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OBJECTIVE:To study the effect of Elemene emulsion combined with ganciclovir on invasive ability and TIMP-1 mRNA expression in C6 brain tumor stem cells. METHODS:Cells with 3 generations were isolated and cultured from rats'C6 ma-lignant glioma cell lines,and immunocytochemistry was adopted to detect the protein expressions of stem cell markers CD133,Nes-tin. 20 rats were randomly divided into blank control group(normal saline),ganciclovir group(intragastrically administrated,216 mg/kg),Elemene emulsion injection group (intravenously injected in tail,36 mg/kg) and combination group (intragastrically ad-ministrated 216 mg/kg of ganciclovir+intravenously injected 36 mg/kg of Elemene emulsion injection in tail),5 in each group, once every day,for 10 d. After 2 h of last administration,the blood of rats in each group was collected,serum was isolated and co-cultured 24 h with C6 BTSCs in in vitro culture medium. Boyden invasion test was used to detect the invasive ability of C6 BTSCs,and real-time quantitative polymerase chain reaction(RT-PCR)method was used to determine the TIMP-1 mRNA expres-sion of C6 BTSCs. RESULTS:Cells with 3 generations had obvious protein expressions in CD133 and Nestin,indicating they were BTSCs. Compared with blank control group,the cell invasion rates of C6 BTSCs in ganciclovir group,Elemene emulsion in-jection group and combination group were obviously decreased,TIMP-1 mRNA expression was obviously enhanced,with statisti-cal significances(P<0.05). Compared with ganciclovir group and Elemene emulsion injection group,the cell invasion rate of C6 BTSCs in combination group was obviously decreased,TIMP-1 mRNA expression was obviously enhanced,with statistical signifi-cances(P<0.05). CONCLUSIONS:Elemene emulsion injection combined with ganciclovir can obviously inhibit the cell invasion of C6 BTSCs,the mechanism may be associated with promoting the TIMP-1 generation,and combination use shows better effect than single use.