Blepharochalasis: Possibly associated with matrix metalloproteinases

Blepharochalasis is a rare disorder characterized by recurrent episodic swelling of eyelids, eventually leading to atrophy of skin. Although immunological mechanisms may be involved in the degradation of elastic fibers, the pathogenesis is not well characterized. We report a 10‐year‐old Japanese boy with an 11‐month history of the swelling of bilateral upper eyelids with atrophic skin. Attacks of non‐painful swelling in eyelids with erythema have recurred several times a month and lasted for 2–3 days. Histological examination revealed perivascular and interstitial infiltration of lymphocytes in the dermis. Elastica Van Gieson staining showed a marked decrease of elastic fibers throughout the dermis. Results of direct and indirect immunofluorescence analyses were negative. Staining of matrix metalloproteinase (MMP)‐3 and MMP‐9 was observed in and around infiltrating cells in the dermis, suggesting that MMP‐3 and MMP‐9 may play, in part, roles in the development of blepharochalasis, and that inhibitor of MMP may have a possibility of therapeutic application.     

Weight loss-induced calciphylaxis: potential role of matrix metalloproteinases

Calciphylaxis is an uncommon and often devastating syndrome of calcification of small vessels, leading to tissue infarction. The mechanism of how calcium deposits on small vessels is unknown. Recently, metalloproteinase digestion of elastin has been shown to enhance deposition of calcium, suggesting a possible mechanism of calciphylaxis. We describe a case of a patient who developed calciphylaxis after rapid weight loss, but had normocalcemia and normal renal function. She was found to have high levels of matrix metalloproteinases, which may have chemically altered elastin, allowing deposition of calcium on small vessels. Inhibitors of matrix metalloproteinases may be useful in the prevention and treatment of calciphylaxis.     

雷公藤红素联合熊去氧胆酸减轻妊娠肝内胆汁淤积的临床研究

目的探讨雷公藤红素联合熊去氧胆酸减轻妊娠肝内胆汁淤积的疗效。方法选择2016年10月至2018年10月深圳市宝安区沙井人民医院诊疗的80例妊娠肝内胆汁淤积患者为研究对象,按照随机分配原则将其分为观察组和对照组,每组40例患者。观察组患者在对照组基础上增加雷公藤片治疗,对照组患者给予熊去氧胆酸治疗。对两组患者的临床疗效、肝功能指标、基质金属蛋白酶水平、不良反应发生率等进行统计比较。结果观察组患者总有效率(92.5%,37/40)高于对照组患者总有效率(75.0%,30/40),组间差异具有统计学意义(P<0.05)。治疗后,观察组患者的谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆汁酸(THA)水平值均低于对照组患者,组间差异均具有统计学意义(均P<0.05)。治疗后,观察组患者的基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)水平均低于对照组患者,组间差异均具有统计学意义(均P<0.05)。观察组患者不良反应发生率(17.5%,7/40)高于对照组患者不良反应发生率(10.0%,4/40),组间比较差异无统计学意义(P>0.05)。结论雷公藤红素联合熊去氧胆酸治疗妊娠期肝内胆汁淤积的临床疗效良好,能够改善患者的肝功能指标水平,降低患者的基质金属蛋白酶-2、基质金属蛋白酶-9水平,且治疗安全性良好,值得临床推荐。     

Activation of toll‐like receptors 2, 3 or 5 induces matrix metalloproteinase‐1 and ‐9 expression with the involvement of MAPKs and NF‐κB in human epidermal keratinocytes

Please cite this paper as: Activation of toll‐like receptors 2, 3 or 5 induces matrix metalloproteinase‐1 and ‐9 expression with the involvement of MAPKs and NF‐κB in human epidermal keratinocytes. Experimental Dermatology 2010; 19 : e44–e49. Abstract: Toll‐like receptors (TLRs) on epidermal keratinocytes are the first line of defense against microbe invasion, and matrix metalloproteases (MMPs) regulate inflammation, cell migration and wound healing. In this study, we demonstrate that the mRNA and protein expressions of MMP‐1 and MMP‐9 in human epidermal keratinocytes are induced by ligands for TLR2, TLR3 and TLR5 [Pam3CSK4, Poly(I:C) and flagellin, respectively] in a dose‐dependent manner. We also found that the ligands for TLR2, TLR3 and TLR5 activate the MAP kinases, JNK and p38 MAPK, but not ERK1/2. Furthermore, treatment with the ligands for TLR2, TLR3 and TLR5 also induced the degradation of IκB‐α and activated the nuclear translocation of NF‐κB. MMP‐1 induction by the ligands for TLR2, TLR3 and TLR5 was inhibited by pretreatment with BAY11‐7082 (NF‐κB inhibitor) or SP600125 (JNK inhibitor), whereas MMP‐9 expression was inhibited by pretreatment with BAY11‐7082, SP600125 or SB203580. These findings demonstrate that the activation of TLR2, TLR3 or TLR5 induces the expression of MMP‐1 and MMP‐9 in human epidermal keratinocytes. In addition, NF‐κB or JNK mediated the MMP‐1 expression induced by TLR2, TLR3 and TLR5, whereas NF‐κB, JNK or p38 MAPK mediated the MMP‐9 expression induced by TLR2, TLR3 and TLR5.     

Clinical relevance of serum levels of matrix metallopeptidase-2, and tissue inhibitor of metalloproteinase-1 and -2 in patients with malignant melanoma

The interaction and/or balance between matrix metallopeptidase (MMP)-2 and tissue inhibitor of metalloproteinase (TIMP)-2 in vivo may play important roles in the process of tumor growth, invasion and metastasis of malignant melanoma. In this study, we investigated the serum levels and immunohistochemical expression of MMP-2, TIMP-1 and TIMP-2 in patients with melanoma and analyzed the correlation with clinicopathological parameters. The level of serum MMP-2 in patients was significantly higher than that of the control. Moreover, the level of MMP-2 was significantly higher than that of the control in patients who were: (i) female; (ii) pT1 and pT4; (iii) with and without lymph node (LN) metastasis; (iv) in stage I and stage IV; (v) with and without recurrence; and (v) alive and dead. The level of serum TIMP-1 in patients with melanoma was significantly higher than that of the control. Among melanoma patients, the level of TIMP-1 with pT4 was significantly higher for patients who were: (i) pT1 and pT3; (ii) with LN metastasis (vs those without); (iii) in stage IV (vs those in stages I, II and III); and (iv) dead (vs those alive). The level of serum TIMP-2 in patients with melanoma was not different from the control. However, the level of TIMP-2 in patients with pT4 was significantly higher than for patients who were: (i) pT1, pT3 and control; (ii) with LN metastasis (vs those without metastasis and control); (iii) with stage IV (vs those in stages I and II and control); (iv) in recurrence (vs control); and (v) dead (vs those alive and control). These results suggest that increased serum levels of TIMP-1 and TIMP-2 reflected the extent of metastatic melanoma lesions, and that serum levels of TIMP-1 may be a new useful marker for melanoma progression.     

Increased matrix metalloproteinase-9 (MMP-9) activity observed in chronic wound fluid is related to the clinical severity of the ulcer

BACKGROUND: The pathology of chronic wounds is often characterized by elevated levels of proinflammatory cytokines [e.g. tumour necrosis factor (TNF)-alpha and interleukin (IL)-1beta], proteases [e.g. matrix metalloproteinases (MMPs)] and neutrophil elastase. MMPs specifically have been implicated by a number of studies as the major protease family responsible for the degradation of key factors critical to the ulcer's ability to heal. OBJECTIVES: To assess individual MMPs in chronic wound fluid (CWF) in order to develop improved treatments for chronic ulcers. METHODS: Collagen type I and IV zymography, immunoprecipitation followed by a substrate activity assay, and an indirect enzyme-linked immunosorbent assay were all used to analyse MMP levels in CWF. RESULTS: Our studies demonstrate that there is excessive protease activity in CWF compared with both human serum and acute wound fluid (AWF), which can be specifically attributed to MMPs as determined through a MMP-inhibitor study. Multiple MMPs were immunoprecipitated from the CWF samples and MMP-9 was identified as the predominant protease in CWF, with significantly elevated activity levels in CWF compared with AWF. In addition, the clinical status of the ulcer is directly associated with the amounts of MMP-9 present in the wound fluid. Therefore, this study suggests that higher levels of MMP-9 in chronic wound fluid correlate with a clinically worse wound. CONCLUSIONS: In view of these results, it is hypothesized that a specific inhibitor of MMP-9 could potentially be more therapeutically effective than general MMP inhibitors in modulating chronic ulcers towards a healing state.     

银屑病皮损中基质金属蛋白酶2、Fas和FasL表达

目的 通过检测银屑病患者皮损中基质金属蛋白酶2(MMP-2)、Fas、FasL的表达，探讨MMP-2对银屑病细胞凋亡的影响。方法 采用免疫组化ABC法对银屑病患者皮损中MMP-2、Fas、FasL表达进行检测。结果 银屑病皮损中既可表达MMP-2，又可表达Fas、FasL而存正常皮肤不表达。结论 MMP-2、Fas、FasL参与了银屑病的发病过程。     

地塞米松对瘢痕疙瘩成纤维细胞基质金属蛋白酶1调控的研究

目的：探讨地塞米松对瘢痕疙瘩成纤维细胞基质金属蛋白酶1（又被称为间质胶原酶1）蛋白的分泌合成和mRNA表达的影响。方法：采用酶联免疫吸附实验（ELISA）和Northem杂交技术检测了培养瘢痕疙瘩成纤维细胞基质金属蛋白酶1蛋白的合成分泌和mRNA的表达。结果：培养瘢痕疙瘩成纤维细胞对照组基质金属蛋白酶1蛋白的浓度为（151．02±27．70）pg／mL；加入10^-9M和10^-6M含地塞米松培养基24小时后，基质金属蛋白酶的浓度分别为（121．27±45．57）pg／mL（P〈0．05）和（101．78±35．82）pg／mL（P〈0．01）。加入地塞米松24小时后，培养瘢痕疙瘩成纤维细胞的基质金属蛋白酶1mRNA的表达被显著抑制，经密度扫描定量分析：基质金属蛋白酶1mRNA的表达分别下降了大约36％和53％。结论：地塞米松可抑制培养瘢痕疙瘩成纤维细胞基质金属蛋白酶1蛋白的合成分泌及其mRNA的表达，可能是地塞米松在抗瘢痕疙瘩纤维化作用中，尤其是纤维化形成后，疗效欠佳的原因。     

SIRT1 modulates expression of matrix metalloproteinases in human dermal fibroblasts

Background SIRT1, an NAD⁺‐dependent histone/protein deacetylase, controls a broad range of cellular functions. Objectives We examined if SIRT1 is involved in the regulation of matrix metalloproteinase (MMP) expression in human dermal fibroblasts. Methods We studied the effect of inhibition of SIRT1 by specific inhibitor and small interfering RNA (siRNA) on MMP‐1 and MMP‐3 expression in human dermal fibroblasts. Results Treatment with a potent and selective inhibitor of SIRT1, EX‐527, increased the basal expression levels of MMP‐1 and MMP‐3 proteins. Knockdown of endogenous SIRT1 by siRNA led to increased expression of MMP‐1 and MMP‐3 at both mRNA and protein levels. SIRT1 knockdown also upregulated MMP protein induction caused by an inflammatory cytokine, interleukin (IL)‐1β. Moreover, treatment with a SIRT1 activator, resveratrol, significantly suppressed IL‐1β‐mediated induction of MMP‐1, which was attenuated by pretreatment with EX‐527. Finally, MMP‐1 promoter activity was increased by EX‐527 in cells treated with or without IL‐1β. Conclusions Our findings suggest that SIRT1 exerts a negative regulatory role in the production of MMP‐1 and MMP‐3 in human dermal fibroblasts.     

Serum levels of tissue inhibitor of metalloproteinase-1 and 2 in patients with eosinophilic fasciitis

BACKGROUND: Serum levels of tissue inhibitor of metalloproteinases (TIMPs) have been reported to be elevated in patients with various connective tissue diseases. However, there has been no report that evaluates TIMPs in patients with eosinophilic fasciitis (EF). OBJECTIVES: To determine serum TIMP-1 and TIMP-2 levels in patients with EF and to investigate their clinical significance. METHODS: Immunohistochemical stainings were performed in normal and EF skin samples. Serum TIMP-1 and TIMP-2 levels of 11 patients with EF and 12 healthy individuals were also measured with specific enzyme-linked immunosorbent assays. RESULTS: The fascia of EF patients was stained only by TIMP-1. Serum TIMP-1 levels (mean +/- SD) were significantly higher in EF patients than in healthy individuals (206.3 +/- 65.4 vs. 145.2 +/- 36.2 ng mL(-1), P < 0.01). Serum TIMP-1 levels in EF patients were significantly correlated with serum gamma-globulin and IgG levels (r = 0.86, P < 0.05; r = 0.83, P < 0.005, respectively). CONCLUSIONS: These results suggest that TIMP-1 is involved in the pathogenesis of EF, and that TIMP-1 may be a useful marker for the disease activity as well as serum gamma-globulin or IgG levels.     

Langerhans cells that express matrix metalloproteinase 9 increase in human dermis during sensitization to diphenylcyclopropenone in patients with alopecia areata

BACKGROUND: We know little of the initial events during the sensitization phase of contact allergy in humans. Alopecia areata (AA), a disease of unknown pathogenesis characterized by patchy hair loss, may be treated by inducing contact allergy to diphenylcyclopropenone (DPC), later followed by its topical application. OBJECTIVES: To learn more about the initial events during sensitization in human skin, we studied the early events during induction of contact allergy to DPC in patients with AA. METHODS: DPC 2% and sodium lauryl sulphate (SLS) 4% were applied on the backs of eight patients with AA. Punch biopsies were taken 6 and 24 h after application. The biopsies were snap-frozen and cryostat sections were evaluated with immunohistochemistry using antibodies against CD1a, HLA-DR, CD3, CD54 and matrix metalloproteinase 9 (MMP-9). RESULTS: After 24 h all subjects exhibited erythema on the DPC-treated areas. Histological evaluation of biopsies from these areas showed hydropic degeneration and a significantly increased number of MMP-9+ cells in the dermis (P < 0.0005). The MMP-9+ cells were identified with double immunofluorescence staining as CD1a + Langerhans cells. The expression of the other markers studied remained unaltered irrespective of treatment, including treatment with SLS. CONCLUSIONS: Our findings show that DPC induces an irritant reaction leading to an increased number of MMP-9+ CD1a+ cells in the dermis during the initial phase of sensitization.     

Quercetin inhibits UV irradiation-induced inflammatory cytokine production in primary human keratinocytes by suppressing NF-κB pathway

Background

Topical flavonoids, such as quercetin, have been shown to reduce ultraviolet (UV) irradiation-mediated skin damage. However, the mechanisms and signaling pathways involved in this protective effect are not clear. UV irradiation leads to activation of two major signaling pathways, namely nuclear factor kappa B (NF-κB) and activator protein-1 (AP-1) pathways. Activation of NF-κB pathway by UV irradiation stimulates inflammatory cytokine expression, whereas activation of AP-1 pathway by UV irradiation promotes matrix metalloproteinase (MMP) production. Both pathways contribute to UV irradiation-induced skin damage, such as photoaging and skin tumor formation.

Objective

To elucidate the underlying mechanism, we examined the effect of quercetin on UV irradiation induced activation of NF-κB and AP-1 pathways.

Methods

Primary human keratinocytes, the major skin cell type subjected to physiological solar UV irradiation, were used to study the effects of quercetin on UV irradiation-induced signal transduction pathways.

Results

Quercetin decreased UV irradiation-induced NF-κB DNA-binding by 80%. Consequently, quercetin suppressed UV irradiation-induced expression of inflammatory cytokines IL-1β (∼60%), IL-6 (∼80%), IL-8 (∼76%) and TNF-α (∼69%). In contrast, quercetin had no effect on UV irradiation activation of three MAP kinases, ERK, JNK, or p38. Accordingly, induction of AP-1 target genes such as MMP-1 and MMP-3 by UV irradiation was not suppressed by quercetin.

Conclusion

Our data indicate that the ability of quercetin to block UV irradiation-induced skin inflammation is mediated, at least in part, by its inhibitory effect on NF-κB activation and inflammatory cytokine production.     

Akt inhibition up‐regulates MMP1 through a CCN2‐dependent pathway in human dermal fibroblasts

Please cite this article as: Akt inhibition up‐regulates MMP1 through a CCN2‐dependent pathway in human dermal fibroblasts. Experimental Dermatology 2010. Abstract: Akt is a key signalling molecule that was found to be down‐regulated in chronic wounds. Akt blockade has dual antifibrotic effects in human dermal fibroblasts, by up‐regulating matrix metalloproteinase 1 (MMP1) and down‐regulating collagen gene expression (J Invest Dermatol 2008: 128: 1906). The aim of this study was to gain additional insights into the mechanism of MMP1 up‐regulation following Akt blockade. As previous studies showed that CCN2 can be a positive regulator of MMP1, we examined the effects of Akt inhibition on CCN2 expression. Akt blockade using a specific pharmacological inhibitor and Akt siRNA resulted in a significant up‐regulation of CCN2, which correlated with the increase in MMP1. The MMP1 up‐regulation following Akt blockade was partially suppressed by CCN2 siRNA, suggesting that CCN2 is contributing to this effect. Additional experiments showed that CCN2 induces phosphorylation of ERK1/2, Ets1 and c‐Jun. Consistent with the stimulatory role of ERK1/2/Ets1 in the expression of MMP1, the ERK1/2 inhibitor UO126 prevented the phosphorylation of ERK1/2 and Ets1 and completely abrogated the induction of MMP1 after CCN2 overexpression, while having no effect on c‐Jun activation. Taken together these results establish CCN2 as a key regulator of MMP1 induction via activation of the ERK1/2/Ets1 pathway. Down‐regulation of Akt signalling leads to inappropriate activation of the CCN2/MMP1 pathway that may contribute to the pathogenesis of chronic wounds. Coordinate expression of CCN2, Akt and MMP1 could be important for normal wound healing to occur. Thus, targeting these specific proteins may represent a promising approach to the therapy of dysregulated wound healing.     

基质金属蛋白酶-8在寻常型银屑病患者血清及皮损中的表达研究

 目的:探究基质金属蛋白酶-8（MMP-8）在寻常型银屑病（PV）炎症反应中的作用。方法:将20例PV患者根据银屑病面积和严重程度指数（PASI）分为轻度组6例、中度组6例及重度组8例。用ELISA法检测并比较PV患者组及正常对照组(12例)血清中MMP-8浓度;用荧光定量PCR扩增并比较不同病情严重程度组间皮损处以及外观正常的周边皮肤MMP-8相对表达量;用免疫组化法观察MMP-8在皮损部位的分布状况。结果:血清MMP-8浓度比较：轻度组明显低于对照组(t=2.54,P=0.022）、重度组（t=2.69,P=0.020）。PV患者皮损部位MMP-8表达明显高于皮损相邻部位外观正常的皮肤（t=2.91,P=0.009）;重度组PV皮损部位MMP-8表达明显高于轻度组（t=3.45,P=0.005）及中度组（t=2.74,P=0.018）。在PV皮损部位,MMP-8主要表达于表皮基底细胞及棘细胞,而在皮损周边部位,MMP-8在基底细胞中表达较明显。真皮中MMP-8主要表达于小血管处。结论:MMP-8在炎症反应较强的皮损中表达上升,推测其通过调控炎症反应及促进血管新生等参与PV的免疫病理进程。     

Interleukin-6 (IL-6) modulates migration and matrix metalloproteinase function in dermal fibroblasts from IL-6KO mice

BACKGROUND: Interleukin-6-deficient (IL-6KO) mice display significantly delayed cutaneous wound healing characterized by decreased re-epithelialization, granulation tissue and wound closure. Dermal fibroblasts are one of the principal cell types found in granulation tissue and mediate numerous processes during healing. OBJECTIVES: To investigate the effects that IL-6 might have on granulation tissue formation and fibroblast motility. As fibroblast motility is associated with matrix metalloproteinase (MMP) activity, the expression of MMP-2 and the tissue inhibitors of metalloproteinase (TIMP)-1 and -2 were assessed. METHODS: Punch biopsies (4 mm) were performed in the skin of IL-6KO and C57BL/6 mice. The expression of MMP-2, TIMP-1 and -2 in wound tissue was monitored over time. Cellular infiltration and granulation tissue formation was monitored by subcutaneous implantation of polyvinyl alcohol (PVA) sponges. A free-floating collagen lattice model was also used to investigate the direct effects of IL-6 treatment on isolated IL-6KO fibroblasts. The expression of MMP-2, and the inhibitors TIMP-1 and -2, were assessed via real-time quantitative polymerase chain reaction and enzyme-linked immunosorbent assay. RESULTS: IL-6KO wounds showed impaired granulation tissue formation 5 days postwounding and fewer fibroblasts had populated the PVA matrices 7 days after implantation in IL-6KO mice compared with wild-type C57BL/6. The mRNA and protein expression of MMP-2 and TIMP-2 mRNA was increased in IL-6KO mice compared with wild-type mice beyond 1 day postwounding, while the expression of TIMP-1 mRNA was transiently higher in IL-6KO only 3 days postwounding. Treatment of collagen lattices with various concentrations of rmIL-6 again showed a dose-response decrease in mRNA and protein expression of MMP-2 and TIMP-2 protein expression, compared with saline control, while TIMP-1 did not appear to be significantly modulated. CONCLUSIONS: These results indicate that IL-6 influences the function of fibroblasts in wounds, and one mechanism of this regulation may be through the modulation of MMP-2 and TIMP proteins.