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1.
Talanta ; 236: 122830, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635220

RESUMO

A sensitive biosensor that can be used for the determination of matrix metalloproteinase 2 (MMP-2) was proposed. The biosensor was developed by using an excellent self-enhanced nanocomposites as an illuminant and a peptide as a recognition element. For the electrostatic attraction between Ru(bpy)32+ and nitrogen-doped graphene quantum dots (NGQDs), the self-enhanced electrochemiluminescence (ECL) nanocomposites of NGQDs-Ru(bpy)32+-doped silica nanoparticles (NGQDs-Ru@SiO2) were synthesized through a simple sol-gel process. Then, a specific peptide (labeled sulfhydryl) was combined with the self-enhanced ECL nanocomposites (carboxyl in NGQDs) via acylation reaction to obtain the peptide-NGQDs-Ru@SiO2 nanoprobe, which was fabricated onto the gold electrode surface via Au-S bond. The peptide of the ECL nanoprobe was exposed to cleavage in the presence of MMP-2, which caused the signal substance to move farther away from the electrode, leading to a decrease of the ECL signal. The proposed NGQDs-Ru@SiO2-labeled peptide ECL biosensor displayed a lower detection limit of 6.5 pg mL-1 than those of reported ECL methods. The proposed biosensor provided an outlook for future applications in other disease-associated biomarkers.


Assuntos
Técnicas Biossensoriais , Grafite , Neoplasias , Pontos Quânticos , Biomarcadores Tumorais , Técnicas Eletroquímicas , Humanos , Medições Luminescentes , Metaloproteinase 2 da Matriz , Nitrogênio , Dióxido de Silício
2.
Biosens Bioelectron ; 195: 113671, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34624798

RESUMO

The extracellular matrix (ECM) of tumor mediates malignant transformation and distant metastasis with extracellular proteinases, especially the matrix metalloproteinases (MMPs). However, there is no assay method to trace the dynamic content of MMPs in ECM. In this work, we have proposed a strategy by assembling peptide scaffold on ionic nanochannels to monitor the target proteinases. The short peptide unit is designed to induce self-assembly with good stability, biocompatibility and programmability, while ion nanochannels can provide electrochemical response upon the MMP activities. Taking MMP-2 as an example, the peptide unit includes two regions, one for self-assembly and one for bio-recognition, so the assembly region (KLVFF) can self-assemble to nanofiber networks. In the meantime, since the reactive region (PLGVR) has MMP-2 recognition site, the peptide assembly on nanochannel can thus be used for the detection of active MMP-2 in tumor microenvironment, with a wide linear detection range (10 fg/mL-10 ng/mL) and 6.6 fg/mL limit of detection. Moreover, the availability of the established ECM mimic is able to distinguish active MMP-2 from latent proMMP-2 in tumor samples. By designing different peptide units for self-assembly on the ionic nanochannel, the assay platform can be promisingly used for other proteinases in ECM, so this work may provide a useful approach to trace the dynamic content of the MMPs in tumor microenvironment (TEM).


Assuntos
Técnicas Biossensoriais , Neoplasias , Matriz Extracelular , Humanos , Metaloproteinase 2 da Matriz , Metaloproteinases da Matriz , Microambiente Tumoral
3.
Acta Cir Bras ; 36(9): e360904, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34755764

RESUMO

PURPOSE: The protective effect of silibinin on kidney and lung parenchyma during hepatic ischemia/reperfusion injury (IRI) is explored. METHODS: Sixty-three Wistar rats were separated into three groups: sham; control (45 min IRI); and silibinin (200 µL silibinin administration after 45 min of ischemia and before reperfusion). Immunohistochemistry and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) were used to evaluate the expression levels of MMP2, MMP3, MMP9, and TIMP2 on kidney and lung. RESULTS: Comparing sham vs. control groups, confirmed that hepatic IRI increased both renal and lung MMP2, MMP3, MMP9 and TIMP2 expressions starting at 180 min (p<0.001). Comparison of the control vs. silibinin groups showed a statistically significant decrease in the expression levels of MMP2, MMP3, and MMP9 and increase of TIMP2 in kidney and lung parenchyma. The starting point of this decrease was at 120 min after reperfusion, both for kidney and lung parameters, and it was statistically significant at 240 min (p<0.001) for kidney, while silibinin showed a peak of lung protection at 180 min after hepatic reperfusion (p<0.001). CONCLUSIONS: Hepatic IRI causes distant kidney and lung damage, while a statistically significant protective action, both on kidney and lung parenchyma, is conveyed by the intravenous administration of silibinin.


Assuntos
Metaloproteinase 2 da Matriz , Traumatismo por Reperfusão , Animais , Isquemia , Rim , Pulmão , Metaloproteinase 3 da Matriz , Metaloproteinase 9 da Matriz , Ratos , Ratos Wistar , Traumatismo por Reperfusão/tratamento farmacológico , Traumatismo por Reperfusão/prevenção & controle , Silibina
4.
Zhonghua Fu Chan Ke Za Zhi ; 56(10): 705-711, 2021 Oct 25.
Artigo em Chinês | MEDLINE | ID: mdl-34823320

RESUMO

Objective: To investigate the effect and mechanism of tumor necrosis factor α (TNF-α) and its inhibitor etanercept (ETA) on the invasion ability of extravillous trophoblast in patients with unexplained recurrent spontaneous abortion (URSA). Methods: (1) Patients were collected from March to June in 2019. They were divided into the URSA group (n=15) and the normal control group (n=15), according to whether diagnosed with URSA or not. The mRNA expression levels of TNF-α in villi tissue of patients in the two groups were detected by quantitative real-time PCR (qRT-PCR). (2) The mRNA and protein expression levels of matrix metalloproteinase-2 (MMP-2), Slug and CXC chemokine rceptor 4 (CXCR4) in HTR-8/SVneo cells were detected by qRT-PCR or western blot after being stimulated by exogenous TNF-α (0.2, 2, 20 ng/ml) alone or TNF-α along with ETA, or phosphate buffered saline (PBS) as control. (3) The invasion ability of HTR-8/SVneo cells was investigated by transwell test after stimulating by TNF-α alone or TNF-α along with ETA. (4) The mRNA and protein expression levels of MMP-2, Slug and CXCR4 in HTR-8/SVneo cells, which were stimulated by TNF-α (2 ng/ml) alone after nuclear factor-κB (NF-κB) inhibitor, BAY 11-7028, preconditioning, were detected by qRT-PCR or western blot. Results: (1) The mRNA expression level of TNF-α in villi tissue of URSA group (4.10±0.49) was 4.1 times as much as the normal control group (t=10.51, P<0.05). (2) The mRNA and protein expression levels of MMP-2, Slug and CXCR4 in HTR-8/SVneo cells of TNF-α group were significantly lower than those in PBS control group (P<0.05) and those in TNF-α along with ETA group (P<0.05). (3) The invasion ability of HTR-8/SVneo cells in TNF-α group was significantly decreased than PBS group and TNF-α along with ETA group (78±14 vs 373±26 vs 227±44, P<0.05). (4) The mRNA and protein expression levels of MMP-2, Slug and CXCR4 in HTR-8/SVneo cells with BAY 11-7028 preconditioning (mRNA: 1.03±0.10, 1.03±0.06, 1.09±0.08; protein: 1.09±0.03, 1.49±0.03, 1.12±0.03) were significantly higher than without preconditioning after being stimulated by TNF-α (all P<0.05). Conclusions: The expression of TNF-α in the villi of URSA patients is much higher than normal early pregnant women. TNF-α could decrease the capacity of invasion by suppressing the expression of MMP-2, Slug and CXCR4 through NF-κB signaling pathway in extravillous trophoblast cells. While ETA could improve the invasiveness capability of extravillous trophoblast cells through inhibiting the negative effect of TNF-α.


Assuntos
Aborto Espontâneo , Fator de Necrose Tumoral alfa , Linhagem Celular , Movimento Celular , Etanercepte/farmacologia , Feminino , Humanos , Metaloproteinase 2 da Matriz/genética , Gravidez , Trofoblastos
5.
Zhongguo Ying Yong Sheng Li Xue Za Zhi ; 37(6): 616-621, 2021 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-34821094

RESUMO

Objective: To investigate the effects and mechanisms of taurine up-regulated gene 1 (TUG1) in hepatic fibrosis. Methods: According to the literature, the classic hepatic fibrosis model of rats induced by 1%DMN(1ml/kg/d) was established. The rats with hepatic fibrosis and activated hepatic stellate cells (HSC) were divided into model control group, negative control group (transfected with siRNA negative control), siRNA interference group (transfected with TUG1). At the end of the experiment, hematoxylin eosin (HE) staining was used to detect the pathological changes of liver tissue; reverse transcription polymerase chain reaction (RT-PCR) and Western blot were used to determine the expression levels of α-smooth muscle actin (α-SMA), TUG1, collagen I, matrix metalloproteinase-2 (MMP-2), tissue inhibitor of metalloproteinase-1 (TIMP-1), Smad2 and Smad3 in rat liver tissue and activated hepatic stellate cells. Results: Compared with the model control group, the protein and gene levels of TUG1 and α-SMA in the negative control group were increased significantly(P<0.05). The protein and gene levels of TUG1, α-SMA, collagen I, MMP-2, TIMP-1, Smad2 and Smad3 in the liver tissue and activated hepatic stellate cells in the siRNA interference group were decreased (P<0.05) while compared with the blank control group and the negative control group. There were no significant differences in the levels of TUG1, α-SMA, collagen I, MMP-2, TIMP-1, Smad2 and Smad3 in the liver tissue and activated hepatic stellate cells between the control group and the negative control group (P>0.05). Conclusion: TUG1 level is elevated in hepatic fibrosis tissue and activated hepatic stellate cells. Silencing TUG1 may improve the pathological damage of hepatic fibrosis induced by 1% DMN by inhibiting the transforming growth factor(TGF-ß1)/ Smad signaling pathway.


Assuntos
Metaloproteinase 2 da Matriz , Inibidor Tecidual de Metaloproteinase-1 , Actinas/genética , Animais , Células Estreladas do Fígado , Cirrose Hepática/patologia , Metaloproteinase 2 da Matriz/genética , Ratos , Inibidor Tecidual de Metaloproteinase-1/genética , Fator de Crescimento Transformador beta1/genética
6.
Zhongguo Ying Yong Sheng Li Xue Za Zhi ; 37(6): 654-659, 2021 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-34821101

RESUMO

Objective: To investigate the effects and molecular mechanism of ropivacaine hydrochloride on osteosarcoma cell proliferation, invasion, and apoptosis. Methods: The osteosarcoma doxorubicin-resistant cell line (U2OS/DOX) was established by gradually increasing the drug doses. U2OS/DOX cells were treated with ropivacaine hydrochloride at the concentrations of 0, 20, 50 and 100 µg/ml, respectively; as different concentrations treatment groups of ropivacaine hydrochloride. pcDNA3.1 and pcDNA3.1-Livin were transfected into U2OS/DOX cells and then treated with 100 µg/ml ropivacaine hydrochloride, which were defined as ropivacaine hydrochloride 100 µg/ml+pcDNA3.1 group, ropivacaine hydrochloride 100 µg/ml+pcDNA3.1-Livin group. MTT was used to detect the cell proliferation inhibition rate and inhibitory concentration (IC50). Western blot was used to detect the expressions of cyclin-dependent kinase inhibitor 1A (P21) and activated cysteine aspartic protease-3 (Cleaved Caspase-3), E-cadherin, matrix metalloproteinase 2 (MMP-2) and Livin; clone formation experiments were used to detect the number of cell clones formed; flow cytometry was used to detect apoptosis; Transwell was used to detect cell migration and invasion; real-time fluorescent quantitative PCR (RT-qPCR) was used to detect Livin mRNA expression. Results: When the concentration of doxorubicin was more than 1 µg/ml, the proliferation inhibition rate of osteosarcoma cells U2OS was significantly increased in a concentration-dependent manner (P<0.05); when the concentration of doxorubicin was more than 10 µg/ml, the proliferation inhibition rate of osteosarcoma resistant cell U2OS/DOX was significantly increased, and it was dose-dependent (P<0.05). In U2OS/DOX cells treated with ropivacaine hydrochloride, the expressions of P21, Cleaved Caspase-3, and E-cadherin were increased significantly, the expression of MMP-2 was decreased significantly, the cell proliferation inhibition rate was increased significantly, the number of colony formation was decreased significantly, and the cells apoptosis rate was increased significantly, the number of cell migration and invasion was decreased significantly, and the expression of Livin was significantly reduced, in a concentration-dependent manner (P<0.05). Overexpression of Livin partially reversed the inhibitory effect of ropivacaine hydrochloride on proliferation, migration, invasion, and promotion effect on apoptosis of cell U2OS/DOX. Conclusion: Ropivacaine hydrochloride can significantly inhibit the proliferation, migration and invasion of doxorubicin-resistant osteosarcoma cells, and significantly promote osteoma cell apoptosis. The mechanism may be related to Livin.


Assuntos
Neoplasias Ósseas , Osteossarcoma , Apoptose , Neoplasias Ósseas/tratamento farmacológico , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Humanos , Metaloproteinase 2 da Matriz , Ropivacaina/farmacologia
7.
World J Gastroenterol ; 27(39): 6615-6630, 2021 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-34754156

RESUMO

BACKGROUND: Extracellular matrix (ECM) remodeling and stiffening, which are correlated with tumor malignancy, drives tumor development. However, the relationship between ECM remodeling and rat experimental model of 1,2-dimethylhyrazine (DMH)-induced colorectal cancer (CRC) imposed by cold and capsaicin exposure remains unclear. AIM: To explore the effects of cold exposure and capsaicin on ECM remodeling and ECM enzymes in DMH-induced CRC. METHODS: For histopathological analysis, the sections of colon tissues were stained with hematoxylin and eosin, Masson's trichrome, Picrosirius red, and Weigert's Resorcin-Fuchsin to observe the remodeling of collagen and elastin. Additionally, the protein expression level of type I collagen (COL I), type 3 collagen (COL III0, elastin, matrix metalloproteinase (MMP) 1, MMP2, MMP9, and tissue-specific matrix metalloproteinase 1 (TIMP1) was assessed by immunohistochemistry. The messenger RNA (mRNA) levels of COL I, COL III, elastin, and lysyl oxidase-like-2 (LOXL2) in the colon tissues of rats was measured by reverse-transcriptase quantitative polymerase chain reaction. RESULTS: Although no differences were observed in the proportion of adenomas, a trend towards the increase of invasive tumors was observed in the cold and capsaicin group. The cold exposure group had a metastasis rate compared with the other groups. Additionally, abnormal accumulation of both collagen and elastin was observed in the cold exposure and capsaicin group. Specifically, collagen quantitative analysis showed increased length, width, angle, and straightness compared with the DMH group. Collagen deposition and straightness were significantly increased in the cold exposure group compared with the capsaicin group. Cold exposure and capsaicin significantly increased the protein levels of COL I, elastin, and LOXL2 along with increases in their mRNA levels in the colon tissues compared with the DMH group, while COL III did not show a significant difference. Furthermore, in immunohistochemical evaluations, MMP1, MMP2, MMP9, and TIMP1 staining increased in the cold exposure and capsaicin group compared with the DMH group. CONCLUSION: These results suggest that chronic cold and capsaicin exposure further increased the deposition of collagen and elastin in the colonic tissue. Increased COL I and elastin mRNA and protein levels expression may account for the enhanced ECM remodel and stiffness variations of colon tissue. The upregulated expression of the LOXL2 and physiological imbalance between MMP/TIMP activation and deactivation could contribute to the progression of the CRC resulting from cold and capsaicin exposure.


Assuntos
Capsaicina , Matriz Extracelular , Animais , Capsaicina/farmacologia , Carcinogênese , Colo , Metaloproteinase 2 da Matriz/genética , Ratos
8.
Braz Dent J ; 32(4): 83-95, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34787255

RESUMO

This study evaluated the bone repair in surgical defects of rats treated with hyaluronic acid (HA) associated or not with Hevea brasiliensis fraction protein (F-1). Bone defect were created in 15 albino Wistar rats divided into 3 groups (n=5): Control group (1) - blood clot; HA group (2) - 0.5% hyaluronic acid; HAF1 group (3) - 0.1% F-1 protein fraction dissolved in 0.5% hyaluronic acid. After 4 weeks, the animals were euthanized and the bone repair was evaluated through histomorphometric analysis, zymography and immunohistochemistry. The neoformed bone area did not show a significant difference (p = 0.757), but there was a tendency for bone trabeculation to increase in the groups HA and HAF1. For immunohistochemically analysis, there was a difference in vascular endothelial growth factor (VEGF) labeling (p = 0.023), being higher in the groups HA and HAF1 than the control group. No significant difference in bone sialoprotein (BSP) (p = 0.681), osteocalcin (p = 0.954), however, significant difference in platelet endothelial cell adhesion molecule-1 (CD-31) (p = 0.040), with HAF1 group being significantly lower than the control. For zymographic analysis, there was no significant difference for metalloproteinase-2 (MMP-2) (p = 0.068), but there was a tendency to increase MMP-2 in the HA group. Despite the influence on angiogenic factors and the apparent tendency for greater trabeculation in the HA and HAF1 groups, there was no significant difference in the area of ​​newly formed bone tissue in the analyzed period.


Assuntos
Ácido Hialurônico , Látex , Animais , Regeneração Óssea , Metaloproteinase 2 da Matriz , Ratos , Fator A de Crescimento do Endotélio Vascular
9.
Chem Pharm Bull (Tokyo) ; 69(10): 1017-1028, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34602570

RESUMO

Celecoxib, a nonsteroidal anti-inflammatory drug, has been reported to have antitumor and antimetastatic activities, and it has potential for application in cancer treatments. The expression of matrix metalloproteinase (MMP)-2/9 is strongly correlated with cancer malignancy, and inhibition of these MMPs is believed to be effective in improving the antitumor and antimetastatic effects of drugs. We have previously revealed that UTX-121, which converted the sulfonamide of celecoxib to methyl ester, has more potent MMP-2/9 inhibitory activity than celecoxib. Based on these findings, we identified compounds with improved MMP inhibitory activity through a structure-activity relationship (SAR) study, using UTX-121 as a lead compound. Among them, compounds 9c and 10c, in which the methyl group of the p-tolyl group was substituted for Cl or F, showed significantly higher antitumor activity than UTX-121, and suppressed the expression of MMP-2/9 and activation of pro MMP-2. Our findings suggest that compounds 9c and 10c may be potent lead compounds for the development of more effective antitumor drugs targeting MMP.


Assuntos
Antineoplásicos/farmacologia , Desenvolvimento de Medicamentos , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Inibidores de Metaloproteinases de Matriz/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Inibidores de Metaloproteinases de Matriz/síntese química , Inibidores de Metaloproteinases de Matriz/química , Estrutura Molecular , Relação Estrutura-Atividade
10.
Cells ; 10(10)2021 10 17.
Artigo em Inglês | MEDLINE | ID: mdl-34685761

RESUMO

Malignant glioma is one of the most lethal cancers with rapid progression, high recurrence, and poor prognosis in the central nervous system. Fatty acid-binding protein 6 (FABP6) is a bile acid carrier protein that is overexpressed in colorectal cancer. This study aimed to assess the involvement of FABP6 expression in the progression of malignant glioma. Immunohistochemical analysis revealed that FABP6 expression was higher in glioma than in normal brain tissue. After the knockdown of FABP6, a decrease in the migration and invasion abilities of glioma cells was observed. The phosphorylation of the myosin light chain was inhibited, which may be associated with migration ability. Moreover, expression levels of invasion-related proteins, matrix metalloproteinase-2 (MMP-2) and cathepsin B, were reduced. Furthermore, tube formation was inhibited in the human umbilical vein endothelial cells with a decreased concentration of vascular endothelial growth factor (VEGF) in the conditioned medium after the knockdown of FABP6. The phosphorylation of the extracellular signal-regulated kinase (ERK), c-Jun NH2-terminal kinase (JNK), and p65 were also decreased after FABP6 reduction. Finally, the bioluminescent images and immunostaining of MMP-2, cluster of differentiation 31 (CD31), and the VEGF receptor 1 (VEGFR1) revealed attenuated tumor progression in the combination of the FABP6-knocked-down and temozolomide (TMZ)-treated group in an orthotopic xenograft mouse tumor model. This is the first study that revealed the impact of FABP6 on the invasion, angiogenesis, and progression of glioma. The results of this study show that FABP6 may be a potential therapeutic target combined with TMZ for malignant gliomas.


Assuntos
Proteínas de Ligação a Ácido Graxo/antagonistas & inibidores , Hormônios Gastrointestinais/antagonistas & inibidores , Glioblastoma/irrigação sanguínea , Glioblastoma/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Neovascularização Patológica/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Animais , Linhagem Celular Tumoral , Movimento Celular/genética , Células Clonais , Progressão da Doença , Matriz Extracelular/metabolismo , Proteínas de Ligação a Ácido Graxo/genética , Proteínas de Ligação a Ácido Graxo/metabolismo , Hormônios Gastrointestinais/genética , Hormônios Gastrointestinais/metabolismo , Regulação Neoplásica da Expressão Gênica , Glioblastoma/patologia , Humanos , Camundongos Nus , Invasividade Neoplásica , Fosforilação , RNA Interferente Pequeno/metabolismo , Temozolomida/farmacologia , Ensaios Antitumorais Modelo de Xenoenxerto
11.
Int J Pharm ; 609: 121209, 2021 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-34678398

RESUMO

Arsenic trioxide (ATO) is the active ingredient in traditional Chinese medicine, i.e., Arsenic, which has shown excellent therapeutic effects on hepatocellular carcinoma. However, due to its poor tumor distribution and high toxicity, the mass adoption of ATO in clinical applications has been severely impeded. In this study, matrix metalloproteinase 2 (MMP2)-responsive cleaved cell-penetrating peptide (PF) and folate (FA) co-modified liposome coated calcium arsenate nanoparticles (FA/PF-LP-CaAs) were fabricated based on these two considerations: (1) The tumor microenvironment characterized by overexpressed MMP2 in extracellular matrix and folate receptor on the cell membrane can enhance drug accumulation and accelerate endocytosis; (2) leveraging different toxicity of arsenic in different valence states, i.e., AsV can be reduced to more toxic AsIII by glutathione in tumor cells. Furthermore, FA/PF-LP-CaAs could be responsively degraded by the mild acidic tumor environment, and the degraded product could escape from lysosomes after endocytosis. More importantly, in light of the in vivo biodistribution and pharmacodynamic studies, the vehicle was able to accumulate in the tumor efficiently. Also, it was able to exhibit excellent anti-tumor efficacy with minimized side effects when compared to single-modified counterparts. Thus, the novel strategy based on the tumor microenvironment proposed in this work can enhance the tumor-targeting efficiency and intratumor toxicity.


Assuntos
Antineoplásicos , Carcinoma Hepatocelular , Neoplasias Hepáticas , Pró-Fármacos , Antineoplásicos/uso terapêutico , Trióxido de Arsênio/uso terapêutico , Carcinoma Hepatocelular/tratamento farmacológico , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Metaloproteinase 2 da Matriz , Pró-Fármacos/uso terapêutico , Distribuição Tecidual , Microambiente Tumoral
12.
J Chem Inf Model ; 61(10): 5203-5211, 2021 10 25.
Artigo em Inglês | MEDLINE | ID: mdl-34649435

RESUMO

Activatable cell-penetrating peptides (ACPPs) are known to be able to decrease the cytotoxicity of cell-penetrating peptide (CPP)-based drug delivery systems. Furthermore, they can improve the targeting of CPPs when specifically recognized and hydrolyzed by characteristic proteases. A comprehensive and profound understanding of the recognition and hydrolysis process will provide a better design of the ACPP-based drug delivery system. Previous studies have clearly described how ACPPs are recognized and bound by MMPs. However, the hydrolysis mechanism of ACPPs is still unsolved. This work focuses on a proteinase-sensitive cleavable linker of ACPPs (PLGLAG), the key structure for recognition and hydrolysis, trying to determine the mechanism by which MMP-9 hydrolyzes its substrate PLGLAG. The quantum mechanics/molecular mechanics (QM/MM) calculations herein show that MMP-9 proteolysis is a water-mediated four-step reaction. More specifically, it consists of (i) nucleophilic attack, (ii) hydrogen-bond rearrangement, (iii) proton transfer, and finally (iv) amide bond rupture. Considering the reversibility of multistep reaction, the second step (i.e., hydrogen-bond rearrangement) has the highest barrier and is the rate-limiting step in the hydrolysis of PLGLAG. The possible design and improvement of the key P1 and P1' sites are also explored through mutations. The present results indicate that, while the mutations affect the reaction energy barriers and the rate-limiting steps, all mutants considered could be hydrolyzed by MMP-9. To provide further insights, the hydrolysis mechanism of MMP-2, which has a similar hydrolysis process to that of MMP-9 but with different reaction barriers, is also studied and compared. As a result, this work provides detailed insights into the hydrolysis mechanism of ACPPs by MMP-9 and, thus, also possible insights for the development of new strategies for ACPP-based delivery systems.


Assuntos
Peptídeos Penetradores de Células , Metaloproteinase 9 da Matriz , Hidrólise , Metaloproteinase 2 da Matriz , Metaloproteinase 9 da Matriz/metabolismo , Simulação de Dinâmica Molecular
13.
Phytomedicine ; 93: 153791, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34666284

RESUMO

BACKGROUND: Recent advancements in understanding ß-escin action provide basis for new therapeutic claims for the drug. ß-escin-evoked attenuation of NF-κB-dependent signaling, increase in MMP-14 and decrease in COUP-TFII content and a rise in cholesterol biosynthesis could be beneficial in alleviating muscle-damaging processes. PURPOSE: The aim of this study was to investigate the effect of ß-escin on skeletal muscle regeneration. METHODS: Rat model of cardiotoxin-induced injury of fast-twich extensor digitorum longus (EDL) and slow-twich soleus (SOL) muscles and C2C12 myoblast cells were used in the study. We evaluated muscles obtained on day 3 and 14 post-injury by histological analyses of muscle fibers, connective tissue, and mononuclear infiltrate, by immunolocalization of macrophages and by qPCR to quantify the expression of muscle regeneration-related genes. Mechanism of drug action was investigated in vitro by assessing cell viability, NF-κB activation, MMP-2 and MMP-9 secretion, and ALDH activity. RESULTS: In rat model, ß-escin rescues regenerating muscles from atrophy. The drug reduces inflammatory infiltration, increases the number of muscle fibers and decreases fibrosis. ß-escin reduces macrophage infiltration into injured muscles and promotes their M2 polarization. It also alters transcription of muscle regeneration-related genes: Myf5, Myh2, Myh3, Myh8, Myod1, Pax3 and Pax7, and Pcna. In C2C12 myoblasts in vitro, ß-escin inhibits TNF-α-induced activation of NF-κB, reduces secretion of MMP-9 and increases ALDH activity. CONCLUSIONS: The data reveal beneficial role of ß-escin in muscle regeneration, particularly in poorly regenerating slow-twitch muscles. The findings provide rationale for further studies on ß-escin repositioning into conditions associated with muscle damage such as strenuous exercise, drug-induced myotoxicity or age-related disuse atrophy.


Assuntos
Escina , Músculo Esquelético , Animais , Metaloproteinase 2 da Matriz , Mioblastos , Ratos , Regeneração
14.
Rom J Morphol Embryol ; 62(1): 239-247, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34609427

RESUMO

Turner syndrome (TS) is characterized by partial or complete loss of a sexual chromosome, resulting in an incomplete development of the body, gonadic failure, cardiac and renal abnormalities, oro-dental changes, etc. In our study, we proposed to perform a histological and immunohistochemical (IHC) study of the periodontium changes in patients with TS. The biological material under study was represented by fragments of gingival mucosa harvested from 18 patients with TS who presented advanced periodontal lesions and required dental extractions. The fragments of gingival mucosa were processed by the classical histological technique of paraffin inclusion, subsequently the obtained sections being stained by the Hematoxylin-Eosin (HE) and examined under the optical microscope. For the IHC study, there were performed serial sections incubated with anti-cluster of differentiation (CD) 3, anti-CD20 and anti-CD68 antibodies for highlighting immune cells, as well as with anti-matrix metalloproteinase (MMP) 2 and anti-MMP8 antibodies for highlighting MMPs (MMP2 and MMP8) involved in the periodontal tissue lesions. In the present study, during the histological examination, there were observed morphological changes, both in the epithelium and in the gingival mucosa chorion. Epithelial changes consisted in the onset of acanthosis processes, in the thickening of the epithelium due to the increase of the spinous layer, as well as in the parakeratosis phenomenon. In the chorion, there was observed the presence of inflammatory infiltrates in various stages, presence of fibrosis (extended in some cases) and the presence of an important vascularization in some cases, with a high number of immunocompetent cells involved in the inborn immune response, but also in the adaptive one, as well as a more or less intense immunoexpression of MMP2 and MMP8. Our study suggests that TS may contribute to the development of some inflammatory processes in the marginal periodontium.


Assuntos
Síndrome de Turner , Epitélio , Gengiva , Humanos , Metaloproteinase 2 da Matriz , Ligamento Periodontal , Periodonto
15.
Biomater Sci ; 9(22): 7456-7470, 2021 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-34609385

RESUMO

Severe hypoxia in solid tumors limits the efficacy of oxygen (O2)-dependent photodynamic therapy (PDT). The overexpressed heat shock proteins (HSPs) in tumor cells hamper the effect of photothermal therapy (PTT). Herein, a tumor oxygenation-enhanced and ATP-reduced gelatin nanoreactor (MCGPD ∼ RGD NPs) for PDT/PTT-augmented combination cancer therapy is reported. In this nanosystem, the Arg-Gly-Asp (RGD) peptides of MCGPD ∼ RGD NPs can ensure accurate recognition and sufficient accumulation in the tumor site. After accumulation, doxorubicin (DOX) can be released from MCGPD ∼ RGD NPs in a mild acidic tumor microenvironment (TME) for highly efficient chemotherapy. Upon 808 nm laser irradiation, the overexpressed matrix metalloproteinase-2 (MMP-2) in the TME and the heat produced from the PDA coating trigger Gel NP degradation to expose chlorin e6 (Ce6) and Met from the cavity of MCGPD ∼ RGD NPs. The exposed Met elevates the O2 content and reduces ATP production in tumor sites to spur the successful O2-dependent PDT and HSP-mediated PTT. The heat generated by the PDA coating directly kills the tumor cells to ensure PTT and amplifies the chemotherapeutic effect. In vitro and in vivo assays indicate that MCGPD ∼ RGD NPs have excellent ability to promote cell apoptosis and to inhibit tumor growth. Overall, this smart responsive hydrogel nanosystem with hypoxia-relieving capacity and ATP-decreasing performance provides a promising strategy against cancer.


Assuntos
Metformina , Nanopartículas , Neoplasias , Fotoquimioterapia , Trifosfato de Adenosina , Linhagem Celular Tumoral , Humanos , Hipóxia , Metaloproteinase 2 da Matriz , Nanotecnologia , Neoplasias/tratamento farmacológico , Fármacos Fotossensibilizantes/uso terapêutico , Microambiente Tumoral
16.
BMC Musculoskelet Disord ; 22(1): 894, 2021 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-34670524

RESUMO

BACKGROUND: Acrolein is a known pro-inflammatory toxic aldehyde, propagating cellular damage and tissue inflammation in humans and animal models of various diseases. Osteoarthritis (OA) has a significant inflammatory component; however, presence of acrolein in synovial fluid of joints with OA has not been previously reported. The first aim of this study was to evaluate evidence of acrolein in the synovial fluid of dogs with OA as well as in Control joints. The second aim was to determine if evidence of acrolein can be detected in synovial fluid samples that have been in a frozen state for long periods of time. METHODS: In this pilot clinical study, synovial fluid samples were prospectively collected (i.e., New samples) from a single joint of both clinically healthy (New Control, n = 5) and dogs with OA (New OA, n = 16) and frozen until the time of analysis. Additionally, frozen synovial fluid samples from a biobank (i.e., Old samples) were used to evaluate ability to detect evidence of acrolein in long-term stored samples (median of 4.89 years) in Old Control (n = 5) and Old OA (n = 5) samples. Measurements of acrolein in all synovial fluid samples was based on detection of its major protein adduct, N ε - (3-formyl-3, 4-dehydropiperidino)lysine (FDP-lysine), using the western blot method. Synovial fluid matrix metalloproteinase 2 (MMP2) was measured in all samples using the western blot method as a positive control of OA inflammation. RESULTS: Acrolein-lysine adduct was detected in both Control (n = 10) and OA (n = 21) groups in both Old and New samples. Acrolein-lysine adduct and MMP2 were detectable at a lower level in the Old compared to New synovial fluid samples; however, the differences were not statistically significant (p > 0.1). The measured MMP2 levels were significantly higher in the OA compared to Control group samples (p = 0.033), but not for acrolein-lysine adduct (p = 0.30). CONCLUSIONS: This study confirmed evidence of acrolein in canine synovial fluid of both OA and Control groups. Freezing of synovial fluid for up to 5 years does not appear to significantly affect the ability to detect acrolein-lysine adduct and MMP2 in these samples.


Assuntos
Osteoartrite , Líquido Sinovial , Acroleína , Animais , Biomarcadores , Cães , Metaloproteinase 2 da Matriz , Osteoartrite/diagnóstico
17.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 39(5): 510-517, 2021 Oct 01.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-34636197

RESUMO

OBJECTIVES: This study aims to investigate the effect of RhoE expression on the migration and invasion of tongue squamous cell carcinoma (TSCC). METHODS: Forty-eight TSCC cases were selected from the Maxillofacial Surgery Center of Qingdao Municipal Hospital from 2017 to 2019. The expression of RhoE in the specimens (TSCC and adjacent tissues) was detected by immunohistochemistry, and RhoE mRNA and protein were extracted to further detect the expression of RhoE. SCC-4 and CAL-27 cells were selected for in vitro experiments. Transient transfection was used to overexpress RhoE. Real-time fluorescence quantitative PCR (qRT-PCR) and Western blot analyses were conducted to detect the overexpression efficiency. Scratch test and Transwell cell invasion tests were used to detect the migration and invasion ability of TSCC, respectively. The expression levels of Rho-associated coiled-coil-containing protein kinase 1 (ROCK1), matrix metalloproteinase-2 (MMP-2), and matrix metalloproteinase-9 (MMP-9) were detected by Western blot. Experimental data were analyzed by Graphpad prism 8.2.1 software. RESULTS: The expression level of RhoE in TSCC was significantly lower than that in adjacent tissues (P<0.05). The migration and invasion abilities of TSCC were significantly lower than those in the control group (P<0.05). The Western blot showed significantly lower expression levels of ROCK1, MMP-2, and MMP-9 in the experimental group than in the control group (P<0.05). CONCLUSIONS: RhoE expression is low in TSCC. Over expression RhoE in TSCC can significantly decrease its migration and invasion abilities. Hence, RhoE may play an important role in regulating the metastasis and invasion of TSCC and provide a new target for gene therapy.


Assuntos
Carcinoma de Células Escamosas , Neoplasias da Língua , Proteínas rho de Ligação ao GTP/genética , Linhagem Celular Tumoral , Humanos , Metaloproteinase 2 da Matriz , Invasividade Neoplásica , Língua , Quinases Associadas a rho
18.
Int J Mol Sci ; 22(19)2021 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-34638665

RESUMO

Matrix metalloproteinases (MMPs) are key signaling modulators in the tumor microenvironment. Among MMPs, MMP-2 and MMP-9 are receiving renewed interest as validated druggable targets for halting different tumor progression events. Over the last decades, a diverse range of MMP-2/9 inhibitors has been identified starting from the early hydroxamic acid-based peptidomimetics to the next generation non-hydroxamates. Herein, focused 1,2,4-triazole-1,2,3-triazole molecular hybrids with varying lengths and decorations, mimicking the thematic features of non-hydroxamate inhibitors, were designed and synthesized using efficient protocols and were alkylated with pharmacophoric amines to develop new Mannich bases. After full spectroscopic characterization the newly synthesized triazoles tethering Mannich bases were subjected to safety assessment via MTT assay against normal human fibroblasts, then evaluated for their potential anticancer activities against colon (Caco-2) and breast (MDA-MB 231) cancers. The relatively lengthy bis-Mannich bases 15 and 16 were safer and more potent than 5-fluorouracil with sub-micromolar IC50 and promising selectivity to the screened cancer cell lines rather than normal cells. Both compounds upregulated p53 (2-5.6-fold) and suppressed cyclin D expression (0.8-0.2-fold) in the studied cancers, and thus, induced apoptosis. 15 was superior to 16 in terms of cytotoxic activities, p53 induction, and cyclin D suppression. Mechanistically, both were efficient MMP-2/9 inhibitors with comparable potencies to the reference prototype hydroxamate-based MMP inhibitor NNGH at their anticancer IC50 concentrations. 15 (IC50 = 0.143 µM) was 4-fold more potent than NNGH against MMP-9 with promising selectivity (3.27-fold) over MMP-2, whereas 16 was comparable to NNGH. Concerning MMP-2, 16 (IC50 = 0.376 µM) was 1.2-fold more active than 15. Docking simulations predicted their possible binding modes and highlighted the possible structural determinants of MMP-2/9 inhibitory activities. Computational prediction of their physicochemical properties, ADMET, and drug-likeness metrics revealed acceptable drug-like criteria.


Assuntos
Ácidos Hidroxâmicos/farmacologia , Bases de Mannich/farmacologia , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Inibidores de Metaloproteinases de Matriz/farmacologia , Triazóis/farmacologia , Antineoplásicos/farmacologia , Células CACO-2 , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Humanos , Micro-Ondas , Simulação de Acoplamento Molecular , Transdução de Sinais/efeitos dos fármacos , Relação Estrutura-Atividade
19.
Int J Mol Sci ; 22(19)2021 Sep 24.
Artigo em Inglês | MEDLINE | ID: mdl-34638621

RESUMO

Previously, we showed that mice treated with cyclophosphamide (CTX) 4 days before intravenous injection of breast cancer cells had more cancer cells in the lung at 3 h after cancer injection than control counterparts without CTX. At 4 days after its injection, CTX is already excreted from the mice, allowing this pre-treatment design to reveal how CTX may modify the lung environment to indirectly affect cancer cells. In this study, we tested the hypothesis that the increase in cancer cell abundance at 3 h by CTX is due to an increase in the adhesiveness of vascular wall for cancer cells. Our data from protein array analysis and inhibition approach combined with in vitro and in vivo assays support the following two-prong mechanism. (1) CTX increases vascular permeability, resulting in the exposure of the basement membrane (BM). (2) CTX increases the level of matrix metalloproteinase-2 (MMP-2) in mouse serum, which remodels the BM and is functionally important for CTX to increase cancer abundance at this early stage. The combined effect of these two processes is the increased accessibility of critical protein domains in the BM, resulting in higher vascular adhesiveness for cancer cells to adhere. The critical protein domains in the vascular microenvironment are RGD and YISGR domains, whose known binding partners on cancer cells are integrin dimers and laminin receptor, respectively.


Assuntos
Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Ciclofosfamida/farmacologia , Metaloproteinase 2 da Matriz/sangue , Microambiente Tumoral/efeitos dos fármacos , Animais , Membrana Basal/efeitos dos fármacos , Neoplasias da Mama/enzimologia , Neoplasias da Mama/patologia , Permeabilidade Capilar/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Feminino , Humanos , Integrina beta1/metabolismo , Neoplasias Pulmonares/irrigação sanguínea , Neoplasias Pulmonares/enzimologia , Neoplasias Pulmonares/secundário , Masculino , Camundongos , Camundongos Knockout , Domínios Proteicos , Microambiente Tumoral/fisiologia
20.
Shanghai Kou Qiang Yi Xue ; 30(4): 402-405, 2021 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-34693434

RESUMO

PURPOSE: To investigate the changes of α-smooth muscle actin (α-SMA), type Ⅰ and type Ⅲ collagen in gingival tissue and expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) in gingival crevicular fluid under orthodontic force. METHODS: Seventy-four patients undergoing orthodontic treatment in Affiliated Stomatological Hospital of Nanchang University from April 2018 to April 2019 were enrolled, and randomly divided into three groups. Group A(n=24) received the treatment under 0 g of orthodontic force, group B (n=25) under 75 g of orthodontic force, and group C(n=25) under 150 g of orthodontic force. At the baseline and 4th week of treatment, the expression levels of α-SMA, type I collagen and type Ⅲ collagen in gingival tissues were detected by immunohistochemical staining. At the baseline, the 2nd, and 4th week of treatment, the expression levels of MMP-2 and TIMP-2 in gingival crevicular fluid were detected by enzyme-linked immunosorbent assay. Then the correlation between different orthodontic force and levels of α-SMA, type Ⅰ collagen, type Ⅲ collagen, MMP-2 and TIMP-2 was analyzed using Spearman correlation analysis. Statistical analysis was completed by SPSS 19.0 software package. RESULTS: At the 2nd and 4th week of treatment, the expression levels of MMP-2 and TIMP-2 in gingival crevicular fluid among three groups were the lowest in group A, followed by group B and group C(P<0.05). At the 4th week of treatment, the levels of α-SMA, type Ⅰ and type Ⅲ collagen in gingival tissues among three groups were the lowest in group A, followed by group B and group C(P<0.05). The orthodontic force was positively correlated with expression levels of α-SMA, type Ⅰ collagen, type Ⅲ collagen, MMP-2 and TIMP-2(P<0.05). CONCLUSIONS: The expression levels of MMP-2 and TIMP-2 in gingival crevicular fluid and myofibroblast are related to the changes of orthodontic force, which may play an important role in the reconstruction of periodontal tissue during orthodontic treatment.


Assuntos
Metaloproteinase 2 da Matriz , Inibidor Tecidual de Metaloproteinase-2 , Actinas , Colágeno Tipo III , Líquido do Sulco Gengival , Humanos , Metaloproteinase 8 da Matriz , Inibidor Tecidual de Metaloproteinase-1
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