Bioactive ingredients in activated platelet-rich plasma derived from cord blood and adult blood via Luminex technology: a comparative study / 中国输血杂志

【Objective】 To compare the bioactive ingredients in activated platelet-rich plasma (PRP) from cord blood and adult blood, explore its reasonable indicators reflecting the inflammatory regulation ability, in order to guide the preparation. 【Methods】 PRP was prepared and activated from 63 healthy adults (31 males, 32 females) and 61 neonates (30 males, 31 females), and 20 cytokines were measured using Luminex technology for assessing the age- and sex-based bioactive differences of PRP. High-sensitivity C-reactive protein(hs-CRP), procalcitonin and MMPs/TIMPs from each sample were measured for their correlations with the 10 inflammation-related cytokines. 【Results】 The activated cord blood PRP released 10 growth factors and chemokines more than the adult blood PRP, whereas IGF-1, HGF and 8 pro-inflammatory cytokines lower than the latter. Most cytokines of adult PRP were more in females than in males (P<0.05), except for IGF-1 and HGF, which showed no difference by gender. Compared with hs-CRP and PCT, MMPs/TIMPs ratio was more closely related with the inflammation-related cytokines, which can reflect the inflammatory regulation of PRP. 【Conclusion】 Due to the lower immunocompetence and no age or gender disturbance, cord blood PRP has the rational MMPs/TIMPs ratio and more cytokines which promote the inflammation and wound healing.

Effect of chitosan nanoparticles on microtensile bond strength of resin composite to dentin: an in vitro study / Efeito de nanopartículas de quitosana na resistência a união por microtração da resina composta à dentina: um estudo in vitro

Objective: The purpose of this study was to evaluate the effect of chitosan nanoparticles on microtensile bond strength of resin composite to dentin using self etch adhesive after aging. Material and Methods: A total number of 90 freshly extracted, sound human molar teeth. Flat tooth surface was gained after cut of the occlusal surface. Three main groups according to pretreatment of dentin before adhesive application; 0.2 % chitosan, 2.5 % chitosan and no treatment control group. Universal self etch adhesive were applied according to manufacture instruction and 4 mm of Feltik Z250 xt composite. Storage of specimens for 1 day, 3 months and 6 months in 37O C distilled water. After that, the tooth was sectioned to beams of 1 mm x8 mm sticks for microtensile bond strength test using universal testing machine. Scanning electron microscope (SEM) was used to evalute the effect of chitosan nanoparticles on dentin and smear layer. Kruskal-Wallis test was used to compare between the three groups as well as the three aging periods. Dunn's test was used for pair-wise comparisons. The significance level was set at P ≤ 0.05. Results: chitosan 0.2% is statistically significant increase in bond strength than chitosan 2.5% and control in one day group. Three months chitosan 0.2 % groups have statistically significant increase in bond strength than chitosan 2.5%. It was found in 6 months that control and chitosan 0.2 % have statistically significant increase in bond strength than chitosan 2.5%. There was statistically significant difference found between the three studied groups regarding bond strength at different storage times . Conclusion: Microtensile bond strength was influenced by different chitosan concentration. Different aging periods had no effect on the microtensile bond strength without application of chitosan and with application of 2.5% chitosan concentration. (AU)

Introdução: O objetivo deste estudo foi avaliar o efeito das nanopartículas de quitosana na resistência da microtração de união do compósito de resina à dentina usando adesivo autocondicionante após o envelhecimento. Material e Métodos: Foram utilizados um total de 90 dentes molares humanos extraídos e sadios. A superfície plana do dente foi obtida após o corte da superfície oclusal. Os dentes foram divididos em três grupos principais de acordo com o pré-tratamento da dentina e antes da aplicação do adesivo: 0,2% de quitosana, 2,5% de quitosana e nenhum tratamento foi utilizado no grupo controle. O adesivo autocondicionante universal foi aplicado de acordo com as instruções do fabricante e 4 mm de composito Feltik Z250 xt foi inserido. O armazenamento de amostras foi realizado por 1 dia, 3 meses e 6 meses em água destilada a 37 °C. Depois disso, o dente foi seccionado em peças de 1 mm x 8 mm para teste de resistência de união por microtração, utilizando máquina de teste universal. Microscópio eletrônico de varredura (MEV) foi usado para avaliar o efeito das nanopartículas de quitosana na dentina e na camada de smear layer. O teste de Kruskal-Wallis foi utilizado para comparar os três grupos e os três períodos de envelhecimento. O teste de Dunn foi usado para comparação pareada dos grupos. O nível de significância foi estabelecido em P ≤ 0,05. (AU)

Effect of exogenous spermine pretreatment on alleviating renal fibrosis in diabetic nephropathy mice and its related mechanism / 中国应用生理学杂志

To observe the protective effects of exogenous spermine on renal fibrosis induced by diabetic nephropathy (DN) and to explore its mechanism. Twenty-four male C57 mice were randomly divided into control group, type 1 diabetes group (TID) and spermine pretreatment group (TID+Sp, n=8 in each group). TID mice were induced by STZ (60 mg/kg), and TID+Sp mice were pretreated with spermine (5 mg/(kg·d)) for 2 weeks before STZ injection. The mice were killed at the 12th week. The renal function was determined by serum creatinine and urea nitrogen. HE, PAS and Masson staining were used to evaluate renal tissue injury and fibrosis. The expressions of matrix metalloproteinase (MMP-2, MMP-9) and collagen IV (Coll-IV) in the kidney of mice were detected by Western blot. Compared with the control group, the blood glucose (5.67±0.22 vs 28.40±0.57 mmol/L), creatinine (14.33±1.22 vs 30.67±4.73 μmol/L) and urea nitrogen (6.93±4.94 vs 22.00±1.04 mmol/L) in the T1D group were increased significantly (P＜0.05), the glomerular basement membrane was thickened, the collagen was significantly increased, the expressions of MMP-2, MMP-9 and Coll-IV protein were increased (0.57±0.07 vs 1.06±0.20, 47.00±0.04 vs 1.29±0.09 and 0.42±0.16 vs 0.95±0.18,P＜0.05). Exogenous spermine significantly alleviates the above-mentioned changes. Exogenous spermine pretreatment could significantly alleviate renal fibrosis in diabetic mice by regulating the balance between MMPs and collagen.

Effect of allogeneic platelet transfusion on migration and invasion of human lung cancer A549 cells and its mechanism of action / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To observe the effect of allogeneic platelets transfusion on the invasion and metastasis of human lung cancer A549 cells, and to preliminarily explore its mechanism of action. Methods: Eighty-nine patients with advanced lung cancer, who had received platelet transfusion in the Chemotherapy Department of Fourth Hospital of Hebei Medical University between January 2017 and December 2018, were enrolled in this study. The study cells were randomized into Ctrl group (A549 cells co-incubated with culture medium), Before group, and After group (A549 cells co-incubated with plasma Before and After platelet transfusion, respectively). The migration and invasion of A549 cells co-cultured with plasma before and after platelet transfection were detected by Scratch and Transwell experiments. The expression of MMPs, TIMPs and epithelial-mesenchymal transition (EMT) related proteins E-cadherin, N-cadherin and Vimentin, as well as vascular endothelial growth factor (VEGF) and its receptor 2 (VEGFR2) were detected by Western blotting (WB) method. Results: The scratch healing ability of A549 cells in After group was significantly higher than that of Ctrl group and Before group [(73.67±2.60)% vs (58.33±2.33)%, (35.33±2.03) %; P<0.01, vs Ctrl group; P<0.05, vs Before group], and there was also a significant difference between Before group and Ctrl group (P<0.05). The results of cell migration experiment showed that the number of transmembrane cells in After group was significantly higher than that in Ctrl group and Before group [(69.67±7.84) vs (18±2.08) and (39.33±2.03), all P<0.01]. The cell invasion experiment showed that the number of transmembrane cells in After group was significantly higher than that in Ctrl group and Before group [(59.34±3.46) vs (18.34±1.56) and (37.58±2.79), all P<0.01]. When A549 cells were co-incubated with plasma before and after platelet transfusion for 48 h, it was found that the expressions of MMP9 and MMP2 were increased (P<0.05), while their inhibitors TIMP1 and TIMP2 were decreased (P<0.01); the expressions of EMT-related proteins N-cadherin and Vimentin were increased (P<0.05), but E-cadherin was decreased (P<0.01); the expressions of angiogenesis related proteins VEGF and VEGFR2 were increased (P<0.05). Conclusion: Alloplatelets transfusion can promote the invasion and metastasis of lung cancer A549 cells, which may be realized by regulation of the expressions of EMT, metallomatrix protease and vascular growth factor-related proteins.

Protective Effect and Mechanism of Tadehaginoside on Hepatic Fibrosis Model Mice Induced by Carbon Tetra- chloride / 中国药房

OBJECTIVE：To study the protective effect of tadehaginoside（TA）on liver fibrosis model mice induced by carbon tetrachloride（CCl4），and to investigate its mechanism. METHODS ：Kunming mice were randomly divided into normal group ， model group ，colchicines group （positive control ，0.2 mg/kg），TA low-dose ，medium-dose and high-dose groups （3，6，12 mg/kg），with 10 mice in each group. Those groups were intraperitoneally injected with 10％ CCl4-olive oil solution （5 mL/kg）to induce liver fibrosis model twice a week ，for consecutive 8 weeks；except that ，the normal group was intraperitoneally injected with olive oil. From 3rd week ，the mice in each administration groups were given relevant medicine intragastrically ，normal group and model group were given constant volume 2％ sodium carboxymethylcellulose solution intragastrically ，once a day ，for consecutive 6 weeks. The contents of ALT ，AST，HA and IL- 6 in serum of mice were test ed by ELISA. The contents of Hyp ， SOD，MDA and GSH-Px in liver tissues were detected by spectrophotometry. mRNA expression of Col- Ⅰ，TIMP-1 and TIMP-2 in liver tissue was detected by RT-PCR. The protein expression of MMP- 2 and TGF-β1 in liver tissue was detected by Western blotting. RESULTS ： Compared with normal group，the contents of ALT，AST，HA，IL-6，MDA and Hyp，the mRNA expression of Col- Ⅰ，TIMP-1 and TIMP- 2，as well as the protein exp ression of MMP- 2 and TGF-β1 were increased significantly ，while the contents of SOD and GSH-Px were decreased significantly （P＜0.01）. Compared with model group，the contents of ALT ，AST，HA，IL-6，MDA and Hyp ，the mRNA expression of Col- Ⅰ，TIMP-1 and TIMP- 2，as well as the protein expression of MMP- 2 and TGF-β1were decreased significantly ，while the contents of SOD and GSH-Px were increased significantly（P＜0.05 or P＜0.01）. CONCLUSIONS ：TA has a significant protective effect on liver tissue and anti-fibrosis effects in liver fibrosis model mice ，the mechanism of which may be associated with inhibiting lipid peroxidation and collagen synthesis ， down-regulating the mRNA expression of Col- Ⅰ，TIMP-1 and TIMP- 2 as well as the protein expression of MMP- 2 and TGF-β1.

Comparison of the effects of triamcinolone acetonide or platelet-rich plasma on expression of extracellular matrix-related genes in equine healthy chondrocytes in vitro / Comparação dos efeitos do acetonido de triancinolona ou plasma rico em plaquetas na expressão de genes relacionados à matriz extracelular de condrócitos in vitro de equinos adultos

ABSTRACT: In healthy cartilage, chondrocytes maintain an expression of collagens and proteoglycans and are sensitive to growth factors and cytokines that either enhance or reduce type II collagen synthesis. In osteoarthritis, pro-inflammatory cytokines, such as IL-6, induce overexpression of metalloproteinases (MMP) and decreasing synthesis of aggrecan. Use of chondroprotectors agents, such as Platelet-Rich Plasma (PRP) and triamcinolone (TA) are alternatives to reduce the progression of joint damage. In this study, we used chondrocytes extracted from metacarpophalangeal joints of healthy horses as the experimental model. Cells were treated in vitro with PRP or TA. No differences were observed between these treatments in comparison to the control group when the expressions of MMP9, MMP13, IL-6 and ACAN genes were evaluated (P<0.05). With these results, we can suggest that the treatments were not deleterious to equine cultured chondrocyte, once they did not stimulate MMPs and IL-6 synthesis or caused changes in ACAN.

RESUMO: Na cartilagem saudável, os condrócitos mantêm a expressão de colágenos e proteoglicanos, sendo sensíveis a fatores de crescimento e citocinas que aumentam ou reduzem a síntese de colágeno tipo II. Na osteoartrite, citocinas pró-inflamatórias, como a IL-6, estimulam a expressão de metaloproteinases (MMP) e reduzem a síntese de agrecano. O uso de condroprotetores, como o Plasma Rico em Plaquetas (PRP) e triancinolona (TA) é uma alternativa para se reduzir a progressão do dano articular. Neste estudo foram usados condrócitos extraídos das articulações metacarpofalangeanas de equinos saudáveis. As células foram tratadas in vitro com TA ou PRP. Não foram observadas diferenças entre os tratamentos comparando-se com o grupo controle quanto à expressão genética de MMP-9, MMP-13, IL-6 e ACAN (p<0,05). Assim, pode-se sugerir que os tratamentos não foram deletérios ao cultivo de condrócitos, uma vez que não estimularam a síntese de MMP e IL-6 e nem causaram alterações no ACAN.

Glycoprotein nonmetastatic melanoma protein B accelerates tumorigenesis of cervical cancer in vitro by regulating the Wnt/ß-catenin pathway

Cervical cancer is one of the most common cancers among women around the world. However, the underlying mechanism involved in cervical cancer progression is incompletely known. In the present study, we determined the role of glycoprotein nonmetastatic melanoma protein B (GPNMB) in tumorigenesis of cervical cancer. According to the GEO database, we found that GPNMB expression was significantly higher in cervical cancer than in normal cervix epithelium. A similar pattern was observed in GPNMB expression in cultured cervical cancer cells and normal cervical epithelial cells. Compared with the control, GPNMB knockdown significantly decreased the proliferation and migration capacity, but enhanced the apoptosis capacity of SiHa and HeLa cells. Additionally, the activity of MMP-2 and MMP-9 were aberrantly increased in SiHa and HeLa cells compared with normal cervical epithelial cells, whereas their activities were strongly inhibited by GPNMB siRNA. Furthermore, Wnt/β-catenin signaling was activated by GPNMB in SiHa and HeLa cells. Increased MMP-2/MMP-9 expression was suppressed by Dkk-1, inhibitor of Wnt/β-catenin signaling, while it was enhanced by stimulator BIO. The proliferation, migration, and apoptosis capacity of HeLa cells were found to be affected by Dkk-1 and BIO to different extents. In conclusion, we demonstrated that GPNMB contributed to the tumorigenesis of cervical cancer, at least in part, by regulating MMP-2/MMP-9 activity in tumor cells via activation of canonical Wnt/β-catenin signaling. This might be a potential therapeutic target for treating human cervical cancer.

Os efeitos das Metaloproteinases da matriz extracelular - MMPS e clorexidina no mecanismo de adesão dentária / The effects of metaloproteinases of the extracellular matrix - MMPs and chlorexidine in the dental adhesion mechanism

Introdução: A adesão da resina composta à dentina ocorre pela formação da camada híbrida. Assim, sua degradação ocasiona a perda da resistência de união na interface resina/dentina, influenciando na longevidade da restauração. Após o condicionamento ácido e aplicação do sistema adesivo na dentina desmineralizada, fibras colágenas não envolvidas por sistema adesivo ficam desprotegidas e suscetíveis ao ataque das metaloproteinases (MMPs). Objetivos: Esta revisão buscou esclarecer o efeito das MMPs na degradação da camada híbrida e os efeitos da clorexidina no processo de adesão. Materiais e métodos: Foi realizada uma revisão da literatura por meio de uma busca bibliográfica nas bases de dados Pubmed/ Medline, Scielo e Google Acadêmico, utilizados estudos publicados nos anos de 2005 a 2018. Foi realizada a busca pelos seguintes descritores: Dentistry, MMPs, Chlorhexidine. Resultados: Estas enzimas, presentes na própria dentina, são reativadas pelo ácido fosfórico ou pelos monômeros ácidos dos adesivos autocondicionantes e iniciam a degradação. A aplicação da clorexidina (CHX) na dentina, após o condicionamento ácido, impede ou retarda a degradação das fibras de colágeno da camada híbrida. Conclusão: Concluiu-se que a ligação adesiva à dentina diminui com o passar dos anos devido à ação das MMPs que degradam o colágeno não infiltrado por monômeros adesivos na parte mais profunda da camada híbrida. Além disso, a clorexidina como inibidor terapêutico em sistemas adesivos convencionais é capaz de inibir as MMPs e assim a ligação adesiva à dentina pode ser mantida estável por um período de tempo mais longo.

Introduction: The adhesion of the composite resin to the dentin occurs by the formation of the hybrid layer. Thus, its degradation causes loss of union resistance on interface resin / dentin interface, directly influencing the longevity of the restoration. After the acid etching and the application of the adhesive system into demineralized dentin, collagen fibers not involved by adhesive system get unprotected and susceptibles to attack by metalloproteinases (MMPs). The enzymes, present in the dentin itself, are rehabilitated by phosphoric acid or by the acids monomers of the self-etching adhesives initiating degradation. The application of chlorhexidine (CHX) in the dentin, after acid conditioning, prevents or slows down the degradation of the collagen fibers of the hybrid layer. This literature review sought to clarify the effect of MMPs on the degradation of the hybrid layer and the effects of chlorhexidine on the adhesion process. It was concluded that the adhesive bonding to dentin decreases with the passage of years due in part to the action of MMPs, which degrade collagen not infiltrated by adhesive monomers in the deepest part of the hybrid layer. In addition, the use of chlorhexidine as a therapeutic inhibitor in conventional adhesive systems is capable of inhibiting the MMPs and thus the adhesive bonding to the dentin can be kept stable for a longer period of time.

Cinobufagin suppresses metastasis and invasion of esophageal carcinoma Kyse-520 cells through FAK/PI3K/AKT pathway / 中国药理学通报

Aim To investigate the effect of cinobufagin on the migration and invasion of esophageal cancer Kyse-520 cells, and to explore the underlying molecular mechanism. Methods The rates of inhibition after treated with different concentrations of cinobufagin 12, 24, 48 h were detected by CCK-8 method, and the changes of cell migration and invasion were observed with wound healing and transwell assay. The mRNA expressions of FAK, Akt, PTEN, VEGF-A, MMP-2 and MMP-9 were detected by quantitative real-time polymerase chain reaction ( RT-qPCR ). The protein expressions of FAK, p-FAK ( Tyr397 ), Akt, p-Akt (Sei473 ), VEGF-A, PTEN, MMP-2 and MMP-9 were measured by Western blot. Results The results of CCK-8 showed that cinobufagin could inhibit the proliferation of Kyse-520 cells in a time- and concentration-dependent manner, and cinobufagin significantly inhibited the cell invasion and migration. Meanwhile, data from RT-qPCR and Western blot suggested that cinobufagin had no significant effect on mRNA and total protein of FAK and Akt, but it reduced the expression of p-FAK(Tyr397), p-Akt(Ser473), VEGF- A, MMP-2 and MMP-9, and increased the PTEN expression. Conclusions Cinobufagin significantly inhibits the invasion and migration of esophageal cancer Kyse-520 cells through inducing PTEN expression and down-regulating FAK/PI3K/AKT pathway.

Circulating Tissue Inhibitor of Metalloproteinase-4 levels are not a Predictor of Preeclampsia in the period between 20 and 25 Weeks of Gestation / Níveis circulantes de inibidor tecidual da metaloproteinase tipo-4 não são um preditor de preeclampsia no período entre 20 e 25 semanas de gestação

Abstract Objective To evaluate whether the circulating level of tissue inhibitor of metalloproteinase- 4 (TIMP-4) in the period between 20 and 25 weeks of gestation is a predictor of preeclampsia. Methods We have performed a case-control study, nested in a prospective study cohort in Ribeirão Preto, in the state of São Paulo, Brazil. Of the 1,400 pregnant women evaluated between 20 and 25 weeks of gestation, 460 delivered in hospitals outside of our institution. Of the 940 pregnant women who completed the protocol, 30 developed preeclampsia. Healthy pregnant women (controls, n = 90) were randomly selected from the remaining 910 participants. From blood samples collected between 20 and 25 weeks of gestation, we performed a screening of 55 angiogenesis-related proteins in 4 cases and 4 controls. The protein TIMP-4 was the most differentially expressed between cases and controls. Therefore, wemeasured this protein in all cases (n = 30) and controls selected (n = 90). Results There were no differences in the plasma TIMP-4 levels of cases compared with controls (1,144 263 versus 1,160 362 pg/mL, respectively; p > 0.05). Conclusion Plasma TIMP-4 levels were not altered at 20 to 25 weeks of gestation, before the manifestation of clinical symptoms; therefore, they are not good predictors of the development of preeclampsia.

Resumo Objetivo Avaliar se o nível de inibidor tecidual de metaloproteinases tipo-4 (TIMP-4, na sigla em inglês) circulante no período entre 20 e 25 semanas de gestação é um preditor de preeclâmpsia. Métodos Foi realizado um estudo caso-controle aninhado em uma coorte de estudo prospectivo em Ribeirão Preto, São Paulo, Brasil. De 1.400 mulheres grávidas avaliadas entre 20 e 25 semanas de gestação, 460 tiveram parto em hospitais fora da nossa instituição. Das 940 gestantes que completaram o protocolo, 30 desenvolveram preeclâmpsia. Gestantes saudáveis (controles, n = 90) foram selecionadas aleatoriamente das 910 participantes restantes. A partir de amostras de sangue coletadas entre 20 e 25 semanas de gestação, foi realizada uma triagem de 55 proteínas relacionadas à angiogênese em 4 casos e 4 controles. A proteína TIMP-4 foi a mais diferentemente expressa entre os casos e os controles; portanto, medimos esta proteína em todos os casos (n = 30) e controles selecionados (n = 90). Resultados Não houve diferenças nos níveis plasmáticos de TIMP-4 nos casos em comparação com os controles (1.144 263 versus 1.160 362 pg/mL, respectivamente; p > 0,05). Conclusão Os níveis plasmáticos de TIMP-4 não foramalterados no período entre 20 e 25 semanas de gestação antes da manifestação dos sintomas clínicos; portanto, não são um bom preditor do desenvolvimento da preeclâmpsia.

MMP2 and MMP9 are Associated with Apical Periodontitis Progression and Might be Modulated by TLR2 and MyD88

Abstract The aim of this study was to evaluate the expression of MMP2 and MMP9 during apical periodontitis (AP) progression in TLR2 (TLR2 KO) and in MyD88 (MyD88 KO) knockout mice compared to wild type (WT) mice. AP was induced in mandibular first molars of TLR2 KO (n= 18), MyD88 KO (n= 18), and WT mice (n= 18). After 7, 21, and 42 days, the animals were euthanized and the jaws were dissected and subjected to histotechnical processing. Subsequent sections were stained by immunohistochemistry and evaluated for detection of MMP2 and MMP9. Statistical analysis of the semi-quantitative analysis of immunohistochemistry was performed using chi-square test (α = 0.05). In the initial periods of AP progression, an increased expression of MMP9 in the TLR2 KO and MyD88 KO mice was observed. In the final periods of AP progression, a reduction of MMP2 expression and an increase of MMP9 expression in the TLR2 KO mice were observed. MMP2 and MMP9 production was modulated for TLR2 and MyD88 during apical periodontitis progression.

Resumo O objetivo deste estudo foi avaliar a expressão de MMP2 e MMP9 durante a progressão da periodontite apical (AP) em camundongos knockout para TLR2 (TLR2 KO) e MyD88 (MyD88 KO) comparados aos camundongos wild type (WT). A AP foi induzida nos primeiros molares inferiores dos camundongos TLR2 KO (n = 18), MyD88 KO (n = 18) e WT (n = 18). Após 7, 21 e 42 dias, os animais foram eutanaziados e as mandíbulas foram dissecadas e submetidas a processamento histotécnico. As lâminas foram coradas por imuno-histoquímica e analisadas para a detecção de MMP2 e MMP9. A análise estatística semi-quantitativa da imuno-histoquímica foi realizada pelo teste qui-quadrado (α = 0,05). Nos períodos iniciais de progressão AP, foi observada uma expressão aumentada de MMP9 nos camundongos TLR2 KO e MyD88 KO. Nos períodos finais de progressão AP, observou-se uma redução da expressão de MMP2 e um aumento da expressão de MMP9 nos camundongos TLR2 KO. A produção de MMP2 e MMP9 foi modulada por TLR2 e MyD88 durante a progressão da periodontite apical.

Inhibition of the Expression of Matrix Metalloproteinases in Articular Chondrocytes by Resveratrol through Affecting Nuclear Factor-Kappa B Signaling Pathway

In the present study, we tried to examine whether resveratrol regulates the expression of matrix metalloproteinases (MMPs) through affecting nuclear factor-kappa B (NF-κB) in articular chondrocytes. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure interleukin-1β (IL-1β)-induced gene expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen. Effect of resveratrol on IL-1β-induced secretion of MMP-3 was investigated in rabbit articular chondrocytes using western blot analysis. To elucidate the action mechanism of resveratrol, effect of resveratrol on IL-1β-induced NF-κB signaling pathway was investigated in SW1353, a human chondrosarcoma cell line, by western blot analysis. The results were as follows: (1) resveratrol inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5, but increased the gene expression of type II collagen; (2) resveratrol reduced the secretion of MMP-3; (3) resveratrol inhibited IL-1β-induced activation (phosphorylation) of inhibitory kappa B kinase (IKK), and thus phosphorylation and degradation of inhibitory kappa Bα (IκBα); (4) resveratrol inhibited IL-1β-induced phosphorylation and nuclear translocation of NF-κB p65. This, in turn, led to the down-regulation of gene expression of MMPs in SW1353 cells. These results suggest that resveratrol can regulate the expression of MMPs through affecting NF-κB by directly acting on articular chondrocytes.

Effects and mechanism of oridonin on invasion and migration abilities of human melanoma A375 cells / 中草药

Objective To study the effects and the mechanism of oridonin on inhibiting the invasion and migration abilities of human melanoma A375 cells. Methods Human melanoma A375 cells were cultured and treated respectively with indicated concentrations of oridonin by cell culture technique. The proliferation rate was detected by CCK-8 method. The migration ability was measured by wound healing assay. The invasion ability was examined by Transwell assay. The adhesion capabilities were evaluated by adhesion assay. The epithelial-mesenchymal transition (EMT) and matrix metalloproteinases (MMPs) related protein expression levels were determined by Western blotting. Results CCK-8 assay showed the median inhibition concentration (IC50) of 48 h was 47.94 μmol/L. Oridonin (5, 10, and 20 μmol/L) inhibited the migration, invasion and adhesion abilities of human melanoma A375 cells in a dose-dependent manner (P < 0.05). After oridonin treatment, the protein expression levels of E-cadherin increased significantly (P < 0.05) and the protein levels of Snail, N-cadherin, vimentin, MMP-2, and MMP-9 decreased significantly (P < 0.05). Conclusion Oridonin inhibits the migration, invasion and adhesion abilities of human melanoma A375 cells. The mechanism may be related with the regulating effects of oridonin on EMT and MMPs.

Segment­ specific expression of MMP/TIMP in the oviduct of llama (Lama glama) and gelatinolytic activity in the oviductal fluid / Expresión segmento específica de MMP/TIMP en el oviducto de llama (Lama glama) y actividad gelatinolítica en el fluido oviductal

Oviductal molecules have the potential to improve the reproductive biotechnologies. In camelids, knowledge and assessment of the oviductal environment are necessary to successfully develop species-specific reproductive technologies, especially because of the camelids reproductive particularities. Among the oviductal factors, the matrix metalloproteinases/tissue inhibitor of matrix metalloproteinases system (MMPs/TIMPs) should be investigated more thoroughly due to their participation in reproductive processes. Consequently, the current study assayed gene and protein expression of MMPs throughout the llama oviduct. MMPs zymogen and active forms in the oviductal fluid were also characterized. MMP2 and MMP9 transcripts were detected in ampulla, isthmus, utero-tubal junction and papilla, being MMP2 and MMP9 2.15 and 1.10 folds higher in papilla than in ampulla, respectively. In addition, differences in immunolocalization of MMP2 and MMP9 between the epithelial mucosa layers of the oviductal segments were observed. The presence of MMPs in the epithelium suggests their secretion into the oviductal lumen. Coincidently, bands of 62 and 94 kDa, corresponding to MMP2 and MMP9 were detected by zymography in the oviductal fluid. Treatment with an exogenous activator (APMA) suggests that they are present as proMMPs. TIMP2 and TIMP1, the specific inhibitors of MMP2 and MMP9, respectively, were expressed in each oviductal segment, indicating a well-regulated control of MMP proteolytic activity in the oviduct. These findings prove that the llama oviduct produces and secretes MMPs into the oviductal lumen, suggesting that these enzymes may have an unknown role in the preparation of the oviductal environment for gametes, fertilization and early embryo development in camelids.

Las moléculas oviductales tienen el potencial para mejorar las biotecnologías reproductivas. En los camélidos, debido a sus peculiares características reproductivas, el conocimiento del ambiente oviductal constituye una herramienta útil para el desarrollo de tecnologías reproductivas específicas para estas especies. Entre los factores oviductales de interés se encuentran las metaloproteasas de matriz (MMPs) y sus inhibidores específicos (TIMPs), los cuales han sido involucrados en diferentes procesos reproductivos. Por estas razones, en este trabajo se caracterizó la expresión génica y proteica de MMP2 y MMP9 en el oviducto de llama. Además, se analizó la presencia de las formas activas e inactivas (zimógenos) de estas enzimas en el fluido oviductal. Se observó que todos los segmentos oviductales, ámpula, istmo, unión útero-tubal y papila, expresan MMP2 y MMP9, siendo los niveles de expresión de MMP2 y MMP9 más elevados en papila respecto a ámpula; 2,15 y 1,10 veces respectivamente. Asimismo, se observaron diferencias en la distribución de las MMPs a nivel de la mucosa entre los segmentos oviductales. Consecuentemente, bandas con actividad gelatinolítica de 62 y 94 kDa, se detectaron en el fluido oviductal, las cuales corresponderían a las formas inactivas de la MMP2 y la MMP9, respectivamente. Los inhibidores específicos de MMP2 y MMP9; TIMP2 y TIMP1, también se detectaron en los segmentos oviductales, indicando su probable participación en la regulación de la actividad proteolítica de las MMPs en el oviducto de llama. En conjunto, los datos de este trabajo demuestran que el oviducto de la llama produce y secreta MMPs al lumen oviductal; sugiriendo que estas enzimas pueden participar en la preparación del ambiente oviductal para la recepción de los gametos, la fecundación y el desarrollo embrionario temprano en camélidos.

Relationship between matrix metalloproteinases and the occurrence and development of ovarian cancer

Ovarian cancer is one of the most malignant genital cancers, with a high mortality rate. Many researchers have suggested that matrix metalloproteinases (MMPs) have remarkably high expression in ovarian cancer tissues. MMPs are considered to be related to the occurrence, development, invasion and metastasis of ovarian cancer. Moreover, some studies have discovered that the unbalance between MMPs and tissue inhibitor of metalloproteinases (TIMPs) are associated with the malignant phenotype of tumors. This review summarizes the latest research progress of MMPs in ovarian cancer. The investigation of MMP mechanism in ovarian cancer will facilitate the development of effective anti-tumor drugs, and thereby improve the survival rate of patients with ovarian cancer.

Effect and mechanism of hesperetin on P-selectin mediated breast cancer MDA-MB-231 metastasis / 中草药

Objective: To study the effect of hesperetin on the migration of P-selectin mediated MDA-MB-231 breast cancer cells and its mechanism. Methods: Using computer virtual docking to evaluate the capacity of hesperetin binding to P-selectin in vitro; MTS test was observed with different concentration of hesperetin or P-selectin on the growth capacity of MDA-MB-231; The effect of hesperetin on P-selectin secretion by activated platelet was detected by Elisa kit; Adhesion experiments examined hesperetin on P-selectin-mediated MDA-MB-231 and endothelial cell adhesion; Transwell experiment was performed to analyze the effect of P-selectin on MDA-MB-231 breast cancer cell migration affected by hesperetin; Western blotting investigated MDA-MB-231 cell surface glycoprotein Mucin-1, Integrin β3, β1 and matrix metalloproteinase expression of MMP-2 and MMP-9 protein expression influenced by hesperetin; impact of hesperetin on MDA-MB-231 cell integrin-matrix metalloproteinase signaling pathway was analyzed to clarify the anti-tumor metastasis mechanism of hesperetin. Results: Hesperetin inhibited P-selectin-induced MDA-MB-231 cell migration and reduced HUVEC-breast cancer cell adhesion. Hesperetin down-regulated the expression of β1 and β3 integrins, and MMP-2 and MMP-9 at protein levels in MDA-MB-231 cells. Conclusion: Hesperetin can inhibit the growth capacity of MDA-MB-231 breast cancer cells, block P-selectin-induced breast cancer MDA-MB-231 tumor cell migration and adhesion, and the mechanism for hesperetin is through competitive P-selectin binding to Mucin-1.Subsequently, hesperetin could block PI3K/AKT/Paxillin/FAK/Src signaling pathway and down-regulate P-selectin mediated Integrins, MMP-2, and MMP-9 expression.

UVB induces morphological changes and the expression of MMPs in human fibroblasts / 基础医学与临床

Objective To observe the changes of telomere length and MMPs level in human fibroblasts induced by UVB, and to explore their roles on skin photoaging .Methods Human skin fibroblasts were extracted and cul-tured.The 5th fibroblasts were irradiated by UVB .The morphology of fibroblasts were microscoped , and the length of telomere and the mRNA expression of COL1a1 and hTERT were detected by RT-qPCR.The expression of MMP-3 and MMP-1 were detected by Western blot .Results The fibroblasts gradually became round , wrinkled and disorderly arranged after 30 mJ/cm2 UVB irradiation for 24 h.The mRNA level of COL1a1 and hTERT and the expression of MMP-3 and MMP-1 were significantly increased after UVB irradiation compared with control , and the length of telomere was shortened .Conclusions UVB may frigger the early process of photoaging by the morphologi-cal changes of human skin fibroblasts and increasing the expression of MMP-3 and MMP-1 .

Advances in research on the prevention and treatment of dental erosion b y MMPs inhibitors / 口腔疾病防治

@#High prevalence of dental erosion has been reported in recent years. The prevention and treatment of dental erosion has therefore attracted lots of attention in dentistry. Matrix metalloproteinases (MMPs) is a type of proteolytic enzyme containing Zn2+ and Ca2+, which can degrade organic matrix in advanced dental erosion. MMPs inhibitors were reported to have the potential to prevent the organic matrix of dentin from degradation, which might provide protective capacity against dental erosion. The present article reviews the mechanism and potential application of MMPs inhibitors in the prevention and treatment of dental erosion.

Effect of oleanolic acid on the activity, secretion and gene expression of matrix metalloproteinase-3 in articular chondrocytes in vitro and the production of matrix metalloproteinase-3 in vivo

In the present study, we tried to examine whether oleanolic acid regulates the activity, secretion and gene expression of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as the production of MMP-3 in the knee joint of rat to evaluate the potential chondroprotective effect of oleanolic acid. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure interleukin-1β (IL-1β)-induced gene expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen. In rabbit articular chondrocytes, the effects of oleanolic acid on IL-1β-induced secretion and proteolytic activity of MMP-3 were investigated using western blot analysis and casein zymography, respectively. The effect of oleanolic acid on in vivo MMP-3 protein production was also examined, after intra-articular injection to the knee joint of rat. The results were as follows: (1) oleanolic acid inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5, but increased the gene expression of type II collagen; (2) oleanolic acid reduced the secretion and proteolytic activity of MMP-3; (3) oleanolic acid suppressed the production of MMP-3 protein in vivo. These results suggest that oleanolic acid can regulate the activity, secretion and gene expression of MMP-3, by directly acting on articular chondrocytes.

Effect of Dracocephalum moldavica total flavones on expression of TGF-β1/Smad signaling pathway and matrix metalloproteinase of atherosclerosis ApoE-/- mice / 中国中药杂志

To investigate the effect and mechanism of Dracocephalum moldovica total flavones (TFDM) on the formation of atherosclerosis ApoE-/- mice induced by high fat diet. A total of 40 SPF 8-week-old male ApoE-/- mice were fed with high fat diet and randomly divided into 5 groups. TFDM high, medium, low-dose group were given 21, 42, 84 mg•kg⁻¹•d⁻¹ by gavage; Simvastatin group was fed with simvastatin 3.5 mg•kg⁻¹•d⁻¹; and model group was given the same dose of normal saline. The other eight male C57BL/6J mice of the same genetic background and age were set up as control group and fed with common diet. All of the groups were intragastrically intervened for 12 weeks. The aortic pathologic changes were observed with HE; qRT-PCR was adopted to detect TGF-β1, Smad2, Smad3, MMP-2 and MMP-9 gene levels in tissues. Compared with model group, HE staining in TFDM group showed obvious relief of aortic atherosclerotic tissue injury; each TFDM group showed inhibition in mRNA expressions of TGF-β1, Smad2, Smad3, MMP-2 and MMP-9. This suggests that TFDM can inhibit atherosclerosis formation, which may be related to the intervention of TGF-β1/Smads signal transduction.