UVB-irradiated human keratinocytes and interleukin-1■ indirectly increase MAP kinase/AP-1 activation and MMP-1 production in UVA-irradiated dermal fibroblasts

Repeated exposure to ultraviolet (UV) irradiation from the sun causes premature skin aging (photoaging). UV irradiation induces the production of matrix metalloproteinases (MMPs) by activating cellular signalling transduction pathways; MMPs are responsible for the degradation and/or inhibition of synthesis of collagenous extracellular matrices in connective tissues.1 Solar UV mainly comprises UVB (wavelength 290 nm to 320 nm) and UVA (320 nm to 400 nm). UVB penetrates the epidermis and…     

Arsenic trioxide regulates the production and activities of matrix metalloproteinases-1, -2, and -9 in fibroblasts and THP-1

Background  The elevated matrix metalloproteinase (MMP) activity is an important cause of chronic wound healing failure. Arsenolite, whose main component is arsenic trioxide (As₂O₃), is a common traditional Chinese medicine wildly used in treating chronic wounds; it can remove necrotic tissue and promote tissue regeneration. This research was designed to evaluate the effects of As₂O₃ on production and activities of MMP-1, MMP-2 and MMP-9, and on regulation of its signal transduction pathway in human skin fibroblasts (HSFb) and human monocyte line (THP-1 cells) that were in an inflammatory state.

Methods  We established three cell models; HSFb activated by TNF-α, THP-1 cells activated by phorbol 12-myristate 13-acetate (PMA) and an HSFb-THP-1 co-culture system. Three cell models was cultured with As₂O₃ for 24 hours. The levels of MMP-1, MMP-2, MMP-9, TNF-α and IL-1β in the cell culture supernatants were assayed by ELISA. The mRNA expressions of MMP-1, MMP-2 and MMP-9 were determined by RT-PCR. The activities of MMP-2 and MMP-9 were tested by Gelatin zymography assays. The phosphorylation levels of ERK1/2 and p38MAPK were assayed by Western blotting.

Results  As₂O₃ inhibited the expression of MMP-1, MMP-2 and MMP-9 mRNA, the secretion and activity of MMP-1, MMP-2 and MMP-9 in HSFb and THP-1 cells in the inflammatory state (P <0.05 and P <0.01 respectively). It also inhibited the secretion of TNF-α and IL-1β in THP-1 cells and in the co-culture system (P <0.05 and P <0.01, respectively). It also decreased the phosphorylation of ERK1/2 and p38 MAPK in HSFb and THP-1 cells (P <0.05 and P <0.01, respectively).

Conclusions  As₂O_3, as a main component of arsenolite, can inhibit the production of MMPs by HSFb and THP-1 cells in an inflammatory state through inhibiting the release of inflammatory factors and the activation of the MAPK cascade pathway. This may be a possible mechanism for arsenolite healing chronic wounds.

     

UVB-irradiated human keratinocytes and interleukin-1α indirectly increase MAP kinase/AP-1 activation and MMP-1 production in UVA-irradiated dermal fibroblasts

Background Solar ultraviolet （UV） irradiation induces the production of matrix metalloproteinases （MMPs） by activating cellular signalling transduction pathways. MMPs are responsible for the degradation and/or inhibition of synthesis of collagenous extracellular matrix in connective tissues. We mimicked the action of environmental ultraviolet on skin and investigated the effects of UVB-irradiated human keratinocytes HaCaT and IL-1α on mitogen activated protein （MAP） kinase activation, c-Jun and c-Fos （AP- 1 is composed of Jun and Fos proteins） mRNA expression and MMP-1 production in UVA-irradiated dermal fibroblasts.     

Advanced glycation end products inhibit production and activity of matrix metalloproteinase-2 in human umbilical vein endothelial cells

Objective It has been reported that advanced glycation end products (AGEs) participate in pathogenesis of accelerated atherosclerosis in diabetes through damaging endothelial function. Matrix metalloproteinases (MMPs) can be synthesized by endothelial cell and contribute to maintain endothelial homeostasis. The present work are aimed at     

Karacoline,identified by network pharmacology,reduces degradation of the extracellular matrix in intervertebral disc degeneration via the NF-κB signaling pathway

Karacoline is a compound found in the plant Aconitum kusnezoffii Reichb.Although Aconitum kusnezoffii Reichb is widely used for the treatment of pain,very few studies have been carried out on the use of karacoline due to its potential toxicity.In this study,we selected key matrix metalloproteinases(MMPs),collagen II,and aggrecan as targets due to their association with intervertebral disc degeneration(IDD).Using these targets,we then used network pharmacology to predict a series of molecules that might exert therapeutic effects on IDD.Of these molecules,karacoline was predicted to have the best effect.Tumor necrosis factor(TNF)-a is known to promote the degeneration of the extracellular matrix in IDD.We therefore applied different concentrations of karacoline(0,1.25,or 12.88 mM)along with 100 ng/mL TNF-a to rat nucleus pulposus cells and found that karacoline reduced the expression of MMP-14 in IDD by inhibiting the nuclear factor(NF)-κB pathway,while collagen II and aggrecan expression was increased.This suggested that extracellular matrix degradation was inhibited by karacoline(P<0.05).Our data therefore reveal a new clinical application of karacoline and provide support for the use of network pharmacology in predicting novel drugs.     

Transforming growth factor-α promotes mouse blastocyst outgrowth and secreti on of matrix metalloproteinases

Objective To study the effect of transforming growth factor-α (TGF-α) on early stage of embryo implantation.Methods Mouse blastocysts were cultured in vitro in medium containing various concentrat ions of TGF-α. Blastocyst implantation capacity was evaluated by calculating the percentage of embryos with attachment or outgrowth. Matrix metalloprotein ases (MMPs) secretion of blastocysts was observed using gelatin zymography. Results There was no significant difference in the percentage of attachment betwee n control and TGF-α treated groups, but the percentage of outgrowth of TGF-α treated groups was significantly higher than that of the control group after 24 h culturing. Gelatin zymography showed that blastocysts cultured in TGF-α treated groups started secreting MMPs earlier than those in the control group.Conclusion TGF-α is involved in regulating the mouse embryo implantation process by promo ting blastocyst outgrowth and secreting matrix matalloproteinases.     

Study on the Effects of FCu-IUD and FICu-IUD on Matrix metalloproteinases in Human Uterine Flushing and Endometrium

It is well know thatmatrix metalloproteinases(MMPs) participate in tissue remodeling duringphysiological and pathological processes,especiallyendometrial cycling,inflammation,tumor invasionand so on[1— 3 ] .We have observed that the activityand kinds of MMPs in the uterine flushing and en-dometrial tissue of FCu- IUD- bearing rabbits wereincreased significantly,the activity of some kindsof MMPs in the FICu- IUD- bearing rabbits weredecreased significantly as compared with the FCu-IUD…     

Role of matrix metalloproteinases in chronic wound healing: diagnostic and therapeutic implications

Matrix metalloproteinases （MMPs） are members of the family of zinc-dependent endopeptidases.     

Effects of matrix metalloproteinase 9 inhibition on the blood brain barrier and inflammation in rats following cardiopulmonary resuscitation

Background Neuroprotective strategies following cardiopulmonary resuscitation （CPR） are an important focus in emergency and critical care medicine. Matrix metalloproteinases （MMPs）, especially MMP9 attracted much attention because of its function in focal brain ischemia/reperfusion injury. In the focal cerebral ischemia model in rats, SB-3CT can suppress the expression of MMP9, relieving brain edema, and there was no studies on global cerebral ischemiareperfusion injury after CPR. Methods One hundred and twenty rats were randomly assigned to sham-operated （n=40）, resuscitation treatment （n=40）, and resuscitation control （n= 40） groups. Sham-operated group rats were anesthetized only and intubated tracheally, while the resuscitation treatment and resuscitation control groups also received cardiac arrest by asphyxiation, In the resuscitation treatment group, SB-3CT was injected intraperitoneally after restoring spontaneous circulation （ROSC）, defined as restoration of supraventricular rhythm and mean arterial pressure （MAP） 〉 60 mm Hg for more than 5 minutes. The resuscitation control group also implemented ROSC without injection of SB-3CT. The rats were executed and samples were taken immediately after death, then at 3, 9, 24, and 48 hours （n=-8）. Brain tissue expression of MMP9 protein, MMP9 mRNA, water content, Evans blue content, TNF-α, IL-1, and IL-6 was measured, and the brain tissue ultramicrostructure studied with electron microscopy. Results In the resuscitation control group, brain tissue expression of MMP9 protein and mRNA, water content, Evans blue content, TNF-α, IL-1, and IL-6 were significantly elevated at 3 hours, and peaked at 24 hours after resuscitation, when compared with the sham-operated group （P 〈0.05）. 1issue ultramicrostructure also changed in the resuscitation control group. By contrast, although all these indexes were increased in the resuscitation treatment group compared with the sham-operated group （P 〈0.05）, they were lower than in the resuscitation control group （P 〈0.05）. Conclusions Expression of MMP9 protein and mRNA, water content, Evans blue content, TNF-α, IL-1, and IL-6 increased in rat brain tissue after CPR, indicating disruption of the blood-brain barrier and excess inflammatory reaction. MMP9 expression was reduced with SB-3CT, resulting in reduced brain injury.     

High mobility group box 1 protein: possible pathogenic link to atrial fibrillation

Atrial fibrillation (AF) is the most common sustained dysrhythmia in clinical practice. The bulk of evidence suggests that inflammatory processes, oxidative stress and matrix metalloproteinase are associated with development of AF. However, these agents may be involved in high mobility group box 1 protein (HMGB1). We hypothesized that HMGB1 may be a possible pathogenic link to AF. A growing body of evidence supports these hypotheses. First, the level of serum HMGB1 is significantly increased in patients with AF including paroxysmal and persistent AF. Second, HMGB1 has been identified as a new pro-inflammatory cytokine in cardiovascular diseases, along with tumor necrosis factor (TNF)-α, interleukin (IL)-6, and C-reactive protein, and there is cross-talk between HMGB1 and inflammatory cytokines. Third, oxidative stress is involved in the release of the pro-inflammatory cytokine, HMGB1, indicating there is cross-talk between oxidative stress and inflammation, and oxidative stress may reinforce the effect of inflammation on the pathogenesis of AF and inflammation may play a more important role in the pathogenesis of AF. Fourth, HMGB1 can promote matrix metalloproteinase-9 upregulation and activation. Fifth, HMGB1 receptors (receptor for advanced glycation end products, Toll-like receptor-2,4) may mediate the atrial structural remodeling or be up-regulated in patients with non-valvular AF. These results suggest that HMGB1 may participate in the pathogenesis of AF and provide a potential target for pharmacological interruption of AF.     

Effects of Danggui Buxue Decoction (当归补血汤) on lipid peroxidation and MMP-2/9 activities of fibrotic liver in rats

Objective:To explore the mechanism of Danggui Buxue Decoction(当归补血汤,DBD) on the liver fibrosis related to hepatic lipid peroxidation and matrix metalloproteinases(MMP) -2/9 activities.Methods: The liver fibrosis in 28 rats was induced by an injection of carbon tetrachloride(CCl_4) and fed with high lipid and low protein diet for 6 weeks,the model rats were randomly divided into the model group and DBD treated group,14 in each group,and another 10 rats as the normal group were observed as well.Rats in the...     

Impact of Combination Use of 0.004% Travoprost and 2% Pilocarpine on Matrix Metalloproteinases Synthesized by Rabbit Ciliary Muscle a Pilot Study

Objective To explore the impact of combination use of prostaglandin analogue and cholinergic agonists on main matrix metalloproteinases （MMPs） synthesized by albino rabbit ciliary muscle. Methods Normal adult albino rabbits were divided into the control group, 2% pilocarpine group, 0.004% travoprost group and travoprost plus pilocarpine group. Two rabbits in the control group were executed after treated with normal saline for one day. Two rabbits were separately executed on the 7th, 14th and 24th day of the treatment in each drug treated group. In each subgroup ciliary muscle band of 4 eyes was taken and made into homogenate. The MMPs activities of 10 subgroups were assayed by zymography. Bands＇ intensity which represents the activity of MMPs was measured by the UltraViolet Illumination system. Results A bright band of MMP-1/2 was showed on each lane at the position corresponding to the molecular weight of 62 kD in the ciliary smooth muscles electrophoresis. When ion Zn and Ca was displaced by＂ MMPs inhibitor EDTA, this bright band disappeared. Compared with the control group, MMP1/2 activity increased by 4.0%, 4.1% and 14.0% after 7, 14 and 24 days ofpilocarpine treatment. Corresponding data was 23.2%, 61.7% and 111.5% in the travoprost group and 49.3%, 68.0% and 88.4% in the travoprost plus pilocarpine group. Conclusions Pilocarpine has little effect on activity of MMP 1 / 2. Travoprost can increase activity of MMP 1/2 gradually. Activity of MMP 1/2 is rapidly increased by pilocarpine combined with travoprost, but shows small change with the prolonged treatment.     

Action mechanisms of complementary and alternative medicine therapies for rheumatoid arthritis

Rheumatoid arthritis(RA) is characterized as a chronic inflammatory disease in joints and concomitant destruction of cartilage and bone.Cartilage extracellular matrix components,such as typeⅡcollagen and aggrecan are enzymatically degraded by matrix metalloproteinases(MMPs) and aggrecanases in RA.Currently,treatments targeting cytokines,including anti-tumor necrosis factor (TNF)αantibodies,soluble TNF receptor,anti-interleukin(IL)-6 receptor antibody, and IL-1 receptor antagonist,are widely used for treating RA in addition to anti-inflammatory agents and disease-modifying antirheumatic drugs(DMARDs),such as methotrexate,but these treatments have some problems,especially in terms of cost and the increased susceptibility of patients to infection in addition to the existence of low-responders to these treatments.Therefore,therapeutics that can be safely used for an extended period of time would be preferable.Complementary and alternative medicines including traditional Chinese medicines(TCM) have been used for the arthritic diseases through the ages.Recently,there are many reports concerning the anti-arthritic action mechanisms of TCM-based herbal formulas and crude herbal extracts or isolated ingredients.These natural herbal medicines are thought to moderately improve RA,but they exert various actions for the treatment of RA.In this review,the current status of the mechanism exploration of natural compounds and TCM-based herbal formulas are summarized,focusing on the protection of cartilage destruction in arthritic diseases including RA and osteoarthritis.     

Involvement of MMP-2 in adriamycin resistance dependent on ERK1/2 signal pathway in human osteosarcoma MG-63 cells

Matrix metalloproteinase-2 (MMP-2) level and the ERK1/2 signal pathway are dependent factors for the growth and metastasis of cancer.However,the impact of MMP-2 in combination with ERK1/2 in tumor patients with drug resistance is unknown.To determine the relationship between MMP-2 and the ERK1/2 signal pathway,we established an adriamycin (ADM)-induced MG-63 (ADM-MG-63) cell line.With the increase of the ERK1/2 pathway blocker PD98059,we detected the expression levels of MMP-2 and p-ERK1/2 by Western blot in ADM-MG-63 cells.In ADM-MG-63 cells transfected with MMP-2-siRNA,the expression of ERK1/2 was detected for understanding the function of the ERK1/2 signal pathway.Three siRNAs for MMP-2 (MMP-2-siRNA) were designed,and the optimal one was selected and tested at different time points of 24,48 and 72 h.Under an ADM-induced condition,ADM-MG-63 cells were finally stable living in the medium of ADM (200 ng/mL).PD98059 could effectively suppress the expression levels of p-ERK1/2 and MMP-2.When the MMP-2 was silenced by using MMP-2-siRNA,the expression of p-ERK1/2 was enhanced.It is con-cluded that MMP-2 may be involved in ADM resistance dependent on ERK1/2 signal pathway,sug-gesting interference in ERK1/2 may be a new method of targeted therapy for tumor resistance.     

Impaired endothelial progenitor cells mobilization: a novel concept for reduce post-ischemia neovascularization in diabetes

Objective The aim of the present study was to explore potential mechanism underlying reduced basal circulating endothelial progenitor cells (EPCs) in diabetes, and to determine how diabetes impairs ischemia-induced mobilization of bone marrow-derived EPCs. Methods Circulating c-Kit /Sca-1 /flk-1 EPCs and CD31bright/Annexin V endothelial microparticles (EMPs) were analyzed by flow cytometry in control and streptozotocin-induced diabetic C57BL/6 mice. Results Compared with nondiabetic controls, diabetic mice had lower level of circulating EPCs, higher level of plasma EMPs, and increased ratio of EMPs to EPCs. Circulating EMPs count and EPCs level were negatively associated with each other. In a hindlimb ischemia model, diabetes suppressed bone marrow-derived EPCs mobilization, accompanied by insufficient upregulaiton of vascular endothelial growth factor (VEGF) and over-activation of matrix metalloproteinases (MMPs) system. As a consequence, diabetic mice showed deficient increase of capillary density and hampered restoration of transcutaneous oxygen pressure (tcpO2) in the ischemic tissue.Concluslon These results suggest that lower circulating EPCs level detected in diabetes may be partly attributed to consumptive loss of EPCs due to increased endothelial damage. Impairment in ischemia-induced EPCs mobilization presents a novel mechanism for defective post-ischemia neovascularization in diabetes.     

The cellular and molecular mechanism of laminin-glycopeptides on anti-metastasis

Objective To further study the anti-metastasis mechanism of laminin-glycopeptides on carcinoma cell proliferation, apoptosis and the secretion of matrix metalloproteinases. Methods Human hepatocellular carcinoma cells in serum free medium were incubated on laminin-coated substrate with or without laminin-glycopeptides at a final concentration of 50 μg/ml. The total number of surviving cells after incubating for the indicated time was assayed by MTT assay. DNA synthesis of the incubated cells was detected by (3)H-TdR incorporation.Cell cycle was analysed by FACS.The mitotic index of Giemsa stained cells was assessed.Cell apoptosis was detected by both FACS and an acridine orange staining method.Matrix metalloproteinase secretion was analysed by gelatin zymography. Results The total number of surviving cells incubated on laminin in the absence of laminin-glycopeptides was significantly larger than that in the presence of laminin-glycopeptides. Laminin promoted (3)H-TdR incorporation of carcinoma cells, decreased the percentage of cells in G1 phase and increased the percentage of cells in S phase.In contrast, laminin-glycopeptides could inhibit the effect of laminin as shown by (3)H-TdR incorporation and cell cycle analysis. The percentage of cells in G2+M phase and the mitotic index among various groups showed no significant difference.Matrix metalloproteinases secretion from cells treated by laminin-glycopeptides was much less compared to that without the treatment by laminin-glycopeptides. Conclusion Laminin may stimulate cell proliferation, while laminin-glycopeptides could significantly inhibit the effect of laminin by inhibiting DNA synthesis and arresting the carcinoma cell cycle from G1 to S phase.These effects may inhibit not only tumor growth of the primary carcinoma, but also the establishment of metastases at ectopic tissues. Laminin-glycopeptides could also inhibit the secretion of matrix metalloproteinases from carcinoma cells and this may contribute to their decreased invasive and metastatic phenotype. This study further revealed the cellular and molecular mechanism of laminin-glycopeptides on anti-metastasis.     

Expression of RECK Gene and MMP-9 in Hilar Cholangiocarcinoma and Its Clinical Significance

Theincidence of pri mary hilar cholangiocarci-noma has anincreasingtrend year by year.The di-agnosis and prognosis of hilar cholangiocarcinomasare poor due to its special location,invasivegrowth,difficulty of early discovery and difficultsurgical operation.At present,some scholars dis-covered a reversion-inducing-cysteine-rich proteinwith Kazal motifs(RECK),which possesses thefunctions uniquelyinhibitingthe expression and ac-tivity of matrix metalloproteinases(MMPs).Thestructure of RECKge…     

Photoprotective effect of the N-terminal 5-mer peptide analog P165 of amyloid precursor protein in human dermal fibroblasts

Background We showed in our previous study that the N-terminal 17-mer peptide of amyloid precursor protein （APP17-mer peptide）,an active peptide segment with trophic and antioxidative effects,protects skin fibroblasts against ultraviolet （UV） damage and downregulates matrix metalloproteinase 1 （MMP-1） expression.The aim of the current study was to explore the protective effects of P165,the N-terminal 5-mer peptide analog of amyloid precursor protein that is resistant to enzymolysis,on UVA-induced damage in human dermal fibroblasts （HDFs）.Methods HDFs were cultured in Dulbecco＆#39;s modified Eagle＆#39;s medium without and with P165 （concentrations were 1,10,and 100 μJmol/L）.Then,15 J/cm2 UVA irradiation was used to obtain the UV-irradiated model.Cell proliferation was analyzed using MTT kit.The collagen type Ⅰ and MMP-1 contents in cell lysate were determined by enzyme-linked immunosorbent assay （ELISA）.Fluorometric assays were performed to detect the formation of intracellular reactive oxygen species （ROS） in the cells.Results P165 significantly protected the HDFs against UVA-induced cytotoxicity.Compared with the UVA-irradiated control,1,10,and 100 μmol/L P165 elevated cell proliferation by 14.98% （P〈0.05）,17.52% （P〈0.01） and 28.34% （P〈0.001）,respectively.Simultaneously,10 and 100 μmol/L P165 increased collagen type Ⅰ content （both P〈0.05）.Moreover,P165 treatment （all concentrations） also markedly suppressed the UVA-induced MMP-1 expression （all P〈0.001）.P165 at 1,10,and 100 μmol/L also reduced UVA-induced ROS generation by 11.27%,13.69% （both P〈0.05）,and 25.48% （P〈0.001）,respectively.Conclusions P165 could protect the HDFs against UVA-induced photodamage,including cytotoxicity,and MMP-1 generation.Furthermore,it also increased the collagen type Ⅰ content in the cells.The inhibitory effect on intracellular ROS generation might be involved in these photoprotective effects.Thus,P165 may be a useful candidate in the prevention and treatment of skin photoaging.