金属蛋白酶及其抑制物在子宫内膜异位症中的表达及其意义

目的 探讨MMPs及TIMPs与子宫内膜异位症发病的关系。方法 采用光镜、免疫组化技术对EM的在位子宫内膜和卵巢异位子宫内膜各17例，子宫内膜腺癌6例，对照组子宫内膜20例分别进行病理组织学观察和MMP－2、MMP－9、MT－MMP和TIMP－1表达的检测。结果 EM的在位子宫内膜和卵巢异位子宫内膜上皮细胞和子宫内膜腺癌细胞MMP－2、MMP－9、MT－MMP表达均显著高于对照组；EM的在位子宫内膜和卵巢异位子宫内膜上皮细胞和子宫内膜腺癌细胞TIMP－1的表达均显著低于对照组。结论 提示EM的发生可能与MMPs及TIMPs等多种因子表达异常相关，而对这些因子的检测可能为临床诊治EM提供新的途径。     

Rheumatoid arthritis: new insights into the role of synovial inflammation in joint destruction

 Rheumatoid arthritis (RA) is characterized by inflammation and proliferation of synovial tissue, leading to degradation of articular cartilage and bone with functional impairment as a result. It has recently become clear that early suppression of synovial inflammation is essential in preventing progressive joint destruction, although inflammation and destruction are in part uncoupled. New insights into the role of matrix metalloproteinases (MMPs), aggrecanase, granzyme B, receptor activator of nuclear factor κB (RANK)–receptor activator of nuclear factor κB ligand (RANKL) interaction, and other factors involved in joint destruction may lead to the development of novel therapies aimed at specific inhibition of cartilage and bone degradation. Correspondence to:P.P. Tak     

Different expression profiles of the lysyl oxidases and matrix metalloproteinases in human ACL fibroblasts after co-culture with synovial cells

Purposes The anterior cruciate ligament (ACL) has poor functional healing response. The synovial tissue surrounding ACL ligament might be a major regulator of the microenvironment in the joint cavity after ACL injury, thus affecting the repair process. Using transwell co-culture, this study explored the direct influence of human synovial cells (HSCs) on ACL fibroblasts (ACLfs) by characterizing the differential expression of the lysyl oxidase family (LOXs) and matrix metalloproteinases (MMP-1, ?2, ?3), which facilitate extracellular matrix (ECM) repair and degradation, respectively. Methods The mRNA expression levels of LOXs and MMP-1, ?2, ?3 were analyzed by semi-quantitative PCR and quantitative real-time PCR. The protein expression levels of LOXs and MMP-1, ?2, ?3 were detected by western blot. Results We found that co-culture resulted in an increase in the mRNAs of LOXs in normal ACLfs and differentially regulated the expression of MMPs. Then we applied 12% mechanical stretch on ACLfs to induce injury and found the mRNA expression levels of LOXs in injured ACLfs were decreased in the co-culture group relative to the mono-culture group. Conversely, the mRNA expression levels of MMPs in injured ACLfs were promoted in the co-culture group compared with the mono-culture group. At translational level, we found that LOXs were lower while MMPs were highly expressed in the co-culture group compared to the mono-culture group. Conclusions The co-culture of ACLfs and HSCs, which mimicked the cell-to-cell contact in a micro-environment, could contribute to protein modulators for wound healing, inferring the potential reason for the poor self-healing of injured ACL.     

Correlation between MMP and TIMP levels and elastic moduli of ascending thoracic aortic aneurysms

Objective

The objective of this preliminary investigation is to determine if there is a relation between the biological levels of matrix metalloproteinases and tissue inhibitor of matrix metalloproteinase (TIMP) and the elastic moduli of the ascending aortic wall in patients with ascending thoracic aortic aneurysms (ATAA). Methods: Circumferential specimens from twelve patients with ATAA were obtained from the greater curvature and their tensile properties (maximum elastic modulus) were tested uniaxially. The levels of MMP1, 2, 3, 8, and 9 as well as TIMP1 and 2 were determined in these aortic wall specimens using MMP/TIMP antibodies array.

Heterogeneity of hepatocellular carcinoma contributes to cancer progression

Hepatocellular carcinoma (HCC) is a highly heterogeneous disease displaying differences in angiogenesis, extracellular matrix proteins, the immune microenvironment and tumor cell populations. Additionally, genetic variations and epigenetic changes of HCC cells could lead to aberrant signaling pathways, induce cancer stem cells and enhance tumor progression. Thus, the heterogeneity in HCC contributes to disease progression and a better understanding of its heterogeneity will greatly aid in the development of strategies for the HCC treatment.     

Vascular Canals in Permanent Hyaline Cartilage: Development,Corrosion of Nonmineralized Cartilage Matrix,and Removal of Matrix Degradation Products

Core areas in voluminous pieces of permanent cartilage are metabolically supplied via vascular canals (VCs). We studied cartilage corrosion and removal of matrix degradation products during the development of VCs in nose and rib cartilage of piglets. Conventional staining methods were used for glycosaminoglycans, immunohistochemistry was performed to demonstrate collagens types I and II, laminin, Ki‐67, von Willebrand factor, VEGF, macrophage marker MAC387, S‐100 protein, MMPs ‐2,‐9,‐13,‐14, and their inhibitors TIMP1 and TIMP2. VCs derived from connective tissue buds that bulged into cartilage matrix (“perichondrial papillae”, PPs). Matrix was corroded at the tips of PPs or resulting VCs. Connective tissue stromata in PPs and VCs comprised an axial afferent blood vessel, peripherally located wide capillaries, fibroblasts, newly synthesized matrix, and residues of corroded cartilage matrix (collagen type II, acidic proteoglycans). Multinucleated chondroclasts were absent, and monocytes/macrophages were not seen outside the blood vessels. Vanishing acidity characterized areas of extracellular matrix degradation (“preresorptive layers”), from where the dismantled matrix components diffused out. Leached‐out material stained in an identical manner to intact cartilage matrix. It was detected in the stroma and inside capillaries and associated downstream veins. We conclude that the delicate VCs are excavated by endothelial sprouts and fibroblasts, whilst chondroclasts are specialized to remove high volumes of mineralized cartilage. VCs leading into permanent cartilage can be formed by corrosion or inclusion, but most VCs comprise segments that have developed in either of these ways. Anat Rec, 300:1067–1082, 2017. © 2016 Wiley Periodicals, Inc.     

A local imbalance between MMP and TIMP may have an implication on the severity and course of appendicitis

Background Matrix metalloproteinases (MMPs) and the tissue inhibitors of MMPs (TIMPs) have been demonstrated to be involved in inflammatory conditions in the intestine. The purpose of this study was to investigate whether the alterations of the MMP/TIMP balance might reflect the course of the inflammatory process in acute appendicitis and if the expression and localisation of MMPs and TIMP is variable in the various clinical manifestations of appendicitis. Materials and methods The study comprises 40 patients (26 men and 14 women) having emergency appendectomy and a control group constituting of 10 patients (5 men and 5 women) having a hemicolectomy for other reasons. MMP and TIMP expressions were assessed and compared in tissue specimens from phlegmonous (n = 15), gangrenous (n = 7), perforated appendicitis (n = 11) and controls with noninflamed appendices (n = 10) by means of enzyme-linked immunosorbent assay technique. Localisation of the enzymes was performed by immunohistochemistry. Results MMP-1 was significantly higher in gangrenous and perforated appendicitis compared with phlegmonous appendicitis and controls (p < 0.05) while MMP-2 was significantly lower in gangrenous appendicitis compared with phlegmonous appendicitis and controls. MMP-2 was also lower in perforated appendicitis when compared with controls (p < 0.01). Elevated expression of MMP-9 was demonstrated in all groups of appendicitis compared with the controls (p < 0.001). Conclusions MMP-9 is the most abundantly expressed MMP of those investigated in inflamed appendix. We postulate that a local imbalance between MMP-9 and TIMP-1 may trigger a perforation. These results suggest that MMPs might be useful as biomarkers of appendices prone to perforation.     

Effect of calcium fluoride on the activity of dentin matrix-bound enzymes

ObjectiveMatrix metalloproteinases (MMPs) and cysteine cathepsins (CCs) are two distinct enzymatic pathways responsible for the degradation of collagen fibrils in demineralized dentin. NaF and KF have been shown to inhibit salivary MMP-2, -9 and CCs. This study investigated the inhibitory effect of calcium fluoride (CaF₂) on the dentin matrix-bound MMPs and CCs.DesignPhosphoric acid (10%)-demineralized dentin beams (1 × 2×6 mm) were incubated at 37 °C in an 1 ml of artificial saliva (AS, control), or AS with 6, 12, 24, 48, 120. 179 and 238 mM F containing CaF₂ (n = 10/group) for 1, 7 and 21 days. All groups were further incubated in AS only for 6 months. Total MMP activity, dry mass loss, CTX and hydroxyproline (HYP) analyses were performed after each incubation. The beams were examined under scanning electron microscopy (SEM). MMP-2 and MMP-9 activities were screened with gelatin zymography. Data were analyzed by using ANOVA and Tukey HSD tests (p = .05).ResultsThe total MMP activity was similar for all groups after 21 days and 6 months. After 21 days, the cumulative mass loss and CTX levels were lower compared to control for the CaF₂ ≥48 and CaF₂≥120 mM, respectively (p < .05). After 6 months, no significant difference was detected in the dry mass loss and CTX compared to the control (p > .05), whereas HYP level was higher with F 24 and 238 mM groups. CaF₂-like minerals were observed on the beams under SEM. There was no gelatinase inhibition in zymography.ConclusionCaF₂ does not prevent the degradation of demineralized dentin matrices due to the catalytic activity of MMPs and CCs.     

Cultivated ginseng suppresses ultraviolet B–induced collagenase activation via mitogen-activated protein kinases and nuclear factor κB/activator protein-1–dependent signaling in human dermal fibroblasts

Cultivated ginseng (CG) (Panax ginseng C.A. Meyer), an herb used in Korean herbal medicine, has been widely used in China and Japan to treat fatigue and to enhance resistance to many diseases. It contains many bioactive constituents, including various ginsenosides that are believed to have antioxidant, immunostimulatory, and antiaging activities. Previous studies have revealed that treatment with Panax ginseng is significantly associated with reduced photoaging, but the underlying mode of action has not been elucidated. In this study, we hypothesized that CG inhibits ultraviolet B (UVB)–induced collagenase activation through mitogen-activated protein kinases (MAPKs) and nuclear factor κB (NF-κB)/activator protein-1 (AP-1)–dependent signaling in human skin fibroblasts. HS68 cells were treated with CG, followed by irradiation with UVB. Those effects were assessed by semiquantitative polymerase chain reaction, Western blotting, and enzymic activity assays. We found that CG increased cell viability and inhibited the production of reactive oxygen species in HS68 cells exposed to UVB irradiation. Pretreatment of HS68 cells with CG inhibited UVB-induced production of matrix metalloproteinase (MMP) 1 and MMP-13. Western blot analysis further revealed that CG markedly suppressed the enhancement of collagen degradation in UVB-exposed HS68 cells. Cultivated ginseng also suppressed UVB-induced activation of NF-κB, c-Jun, and c-Fos and the phosphorylation of MAPKs, which are upstream modulators of NF-κB and AP-1. These results indicate that CG inhibits UVB-induced collagenolytic MMP production by interfering with MAPK/AP-1 and NF-κB signaling and thus may be useful in the prevention and treatment of skin photoaging.