Matrix metalloproteinases in the restorative proctocolectomy pouch of pediatric ulcerative colitis

AIM:To investigate matrix metalloproteinases(MMPs) and their tissue inhibitors(TIMPs) in pouch mucosa of pediatric onset ulcerative colitis(UC).METHODS:In this cross-sectional study,28 patients with pediatric onset UC underwent ileal pouch biopsy 13 years(median) after proctocolectomy.Expression of MMPs-3,-7,-8,-9,-12 and-26 and TIMPs-1,-2 and-3 in samples was examined using immunohistochemichal methods,and another biopsy was used to evaluate the grade of histological inflammation.Two investigators independently graded the immunohistochemical specimens in a semiquantitative fashion,using a scale marking staining intensity as follows:0 = less than 20 positive cells;1 = 20-50 positive cells;2 = 50-200 positive cells;3 = over 20 positive cells.Fecal calprotectin and blood inflammatory markers [serum C-reactive protein(CRP) and erythrocyte sedimentation rate] were determined during a follow-up visit to examine correlations between these markers and the expression of MMPs and TIMPs.RESULTS:Of the 28 patients with pediatric onset UC,nine had not experienced pouchitis,whereas thirteen reported a single episode,and six had recurrent pouchitis(≥ 4 episodes).At the time of the study,six patients required metronidazole.In all of the others,the most recent episode of pouchitis had occurred over one month earlier,and none were on antibiotics.Only four samples depicted no sign of inflammation,and these were all from patients who had not had pouchitis.Two samples were too small to determine the grade of inflammation,but both had suffered pouchitis,the other recurrent.No sample depicted signs of colonic metaplasia.Most pouch samples showed expression of epithelial(e) and stromal(s) MMP-3(e,n = 22;s,n = 20),MMP-7(e,n = 28;s,n = 27),MMP-12(e,n = 20;s,n =24),TIMP-2(e,n = 23;s,n = 23) and MMP-3(e,n = 23;s,n = 28) but MMP-8(e,n = 0;s,n = 1),MMP-9(e,n = 0;s,n = 9) and MMP-26(e,n = 0;s,n = 3) and TIMP-1(n = 0,both) were lacking.In samples with low grade of inflammatory activity,the epithelial MMP-3 and MMP-7 expression was increased(r =-0.614 and r =-0.472,respectively,P < 0.05 in both).MMPs and TIMPs did not correlate with the markers of inflammation,fecal calprotectin,erythrocyte sedimentation rate,or CRP,with the exception of patients with low fecal calprotectin(< 100 μg/g) in whom a higher expression of epithelial MMP-7 was found no differences in MMPor TIMP-profiles were seen in patients with a history of pouchitis compared to ones with no such episodes.Anastomosis with either straight ileoanal anastomosis or ileoanal anastomosis with J-pouch did depict differences in MMP-or TIMP-expression.CONCLUSION:The expression of MMPs pediatric UC pouch in the long-term shares characteristics with inflammatory bowel disease,but inflammation cannot be classified as a reactivation of the disease.

Effects of HDAC4 on IL-1β-induced matrix metalloproteinase expression regulated partially through the WNT3A/β-catenin pathway

Background::Histone deacetylase 4(HDAC4)regulates chondrocyte hypertrophy and bone formation.The aim of the present study was to explore the effects of HDAC4 on Interleukin 1 beta(IL-1β)-induced chondrocyte extracellular matrix degradation and whether it is regulated through the WNT family member 3A(WNT3A)/β-catenin signaling pathway.Methods::Primary chondrocytes(CC)and human chondrosarcoma cells(SW1353 cells)were treated with IL-1βand the level of HDAC4 was assayed using Western blotting.Then,HDAC4 expression in the SW1353 cells was silenced using small interfering RNA to detect the effect of HDAC4 knockdown on the levels of matrix metalloproteinase 3(MMP3)and MMP13 induced by IL-1β.After transfection with HDAC4 plasmids,the overexpression efficiency was examined using Real-time quantitative polymerase chain reaction(qRT-PCR)and the levels of MMP3 and MMP13 were assayed using Western blotting.After incubation with IL-1β,the translocation ofβ-catenin into the nucleus was observed using immunofluorescence staining in SW1353 cells to investigate the activation of the WNT3A/β-catenin signaling pathway.Finally,treatment with WNT3A and transfection with glycogen synthase kinase 3 beta(GSK3β)plasmids were assessed for their effects on HDAC4 levels using Western blotting.Results::IL-1βdownregulated HDAC4 levels in chondrocytes and SW1353 cells.Furthermore,HDAC4 knockdown increased the levels of MMP3 and MMP13,which contributed to the degradation of the extracellular matrix.Overexpression of HDAC4 inhibited IL-1β-induced increases in MMP3 and MMP13.IL-1βupregulated the levels of WNT3A,and WNT3A reduced HDAC4 levels in SW1353 cells.GSK-3βrescued IL-1β-induced downregulation of HDAC4 in SW1353 cells.Conclusion::HDAC4 exerted an inhibitory effect on IL-1β-induced extracellular matrix degradation and was regulated partially by the WNT3A/β-catenin signaling pathway.

Effects of (-)-epigallocatechin-3-gallate on expression of matrix metalloproteinase-1 and tissue inhibitor of metalloproteinase-1 in fibroblasts irradiated with ultraviolet A

Background It is known that ultraviolet irradiation can affect cellular function through a number of signaling pathways ( ) epigallocatechin 3 gallate (EGCG) is the major effective component in green tea and can offer protection from ultraviolet induced damage In this study, we investigated the protective mechanism of EGCG on human dermal fibroblasts damaged by ultraviolet A (UVA) in vitro Methods Transcription factor Jun protein levels were measured by Western blot Matrix metalloproteinase 1 (MMP 1) and tissue inhibitor of metalloproteinase 1 (TIMP 1) mRNA were studied by reverse transcription polymerase chain reaction (RT PCR) analysis in conjunction with computer assisted image analysis MMP 1 and TIMP 1 proteins were quantified by enzyme linked immunosorbent assay (ELISA) Results EGCG decreased transcription activity of Jun protein after induction by UVA Both the mRNA and protein levels of MMP 1 were increased by UVA irradiation, while no significant changes were observed in TIMP 1 levels The ratio of MMP 1 to TIMP 1 showed statistically significant differences compared with the control EGCG decreased the ratio of MMP 1 to TIMP 1 by inhibiting UVA induced MMP 1 expression ( P <0 05) Conclusion EGCG can protect human fibroblasts against UVA damage by downregulating the transcription activity of Jun protein and the expression of MMP 1 The ratio of MMP 1 to TIMP 1, rather than the levels of MMP 1 or TIMP 1 alone, may play a significant role in human skin photodamage

High-performance lung-targeted bio-responsive platform for severe colistin-resistant bacterial pneumonia therapy

Polymyxins are the last line of defense against multidrug-resistant(MDR)Gram-negative bacterial infections.However,this last resort has been threatened by the emergence of superbugs carrying the mobile colistin resistance gene-1(mcr-1).Given the high concentration of matrix metalloproteinase 3(MMP-3)in bacterial pneumonia,limited plasma accumulation of colistin(CST)in the lung,and potential toxicity of ionic silver(Ag+),we designed a feasible clinical transformation platform,an MMP-3 high-performance lung-targeted bio-responsive delivery system,which we named“CST&Ag@CNMS”.This system exhibited excellent lung-targeting ability(>80%in lungs),MMP-3 bio-responsive release property(95%release on demand),and synergistic bactericidal activity in vitro(2-4-fold minimum inhibitory concentration reduction).In the mcr-1+CST-resistant murine pneumonia model,treatment with CST&Ag@CNMS improved survival rates(70%vs.20%),reduced bacteria burden(2-3 log colony-forming unit[CFU]/g tissue),and considerably mitigated inflammatory response.In this study,CST&Ag@CNMS performed better than the combination of free CST and AgNO3.We also demonstrated the superior biosafety and biodegradability of CST&Ag@CNMS both in vitro and in vivo.These findings indicate the clinical translational potential of CST&Ag@CNMS for the treatment of lung infections caused by CST-resistant bacteria carrying mcr-1.

Efficacy of knee-balancing manipulation plus heat-sensitive moxibustion for knee osteoarthritis and its influence on CTX-Ⅰ,TRACP-5b,ADAMTS-4,and MMP-3

Objective To observe the efficacy of knee-balancing manipulation plus heat-sensitive moxibustion in treating knee osteoarthritis(KOA)and its impact on the expression of C-telopeptide of type I collagen(CTX-Ⅰ),tartrate-resistant acid phosphatase 5b(TRACP-5b),A disintegrin and metalloproteinase with thrombospondin motifs 4(ADAMTS-4),and matrix metalloproteinase 3(MMP-3).Methods A total of 134 unilateral KOA patients were randomized into a knee-balancing group,a heat-sensitive moxibustion group,and a joint intervention group.The knee-balancing group received knee-balancing Tuina(Chinese therapeutic massage)manipulation for treatment.The heat-sensitive moxibustion group received heat-sensitive moxibustion treatment.The joint intervention group received the heat-sensitive moxibustion in addition to the knee-balancing manipulation.The intervention period lasted for four weeks.After the treatment,and at the 2-week and 6-week follow-ups,the three groups were assessed using the visual analog scale(VAS)for knee joint pain and Western Ontario and McMaster Universities arthritis index(WOMAC),and clinical efficacy was also evaluated.The enzyme-linked immunosorbent assay was adopted to detect the expression levels of serum CTX-Ⅰ,TRACP-5b,ADAMTS-4,and MMP-3.Results The knee-balancing group had 44 participants,but one dropped out;there was no dropout case among the 44 participants in the heat-sensitive moxibustion group;among the 46 participants in the joint intervention group,two cases dropped out.After the treatment,and at the 2-week and 6-week follow-ups,the total effective rate was found higher in the joint intervention group than in the knee-balancing and heat-sensitive moxibustion groups(P<0.05).Compared with the baseline,the VAS and WOMAC scores and the serum levels of CTX-Ⅰ,TRACP-5b,ADAMTS-4,and MMP-3 decreased significantly in all three groups after treatment and at the 2-week and 6-week follow-ups(P<0.05).At the same three time points,the VAS and WOMAC scores and serum levels of CTX-Ⅰ,TRACP-5b,ADAMTS-4,and MMP-3 were lower in the joint intervention group than in the knee-balancing and heat-sensitive moxibustion groups(P<0.001).Conclusion Either used alone or combined,the knee-balancing manipulation and heat-sensitive moxibustion therapy can improve the symptoms and down-regulate the serum levels of CTX-Ⅰ,TRACP-5b,ADAMTS-4,and MMP-3 in KOA patients,producing durable efficacy;nevertheless,a more significant efficacy can be achieved by combining the two methods.