Effects of HDAC4 on IL-1β-induced matrix metalloproteinase expression regulated partially through the WNT3A/β-catenin pathway / 中华医学杂志(英文版)

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-1β and 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 of β-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-1β downregulated 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-1β upregulated the levels of WNT3A, and WNT3A reduced HDAC4 levels in SW1353 cells. GSK-3β rescued 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.

Expression of cartilage oligomeric matrix protein in the synovial chondromatosis of the temporomandibular joint / 北京大学学报(医学版)

OBJECTIVE@#To detect the expression of cartilage oligomeric matrix protein (COMP) in the synovial chondromatosis of the temporomandibular joint (TMJSC), and to discuss the possible interactions between COMP, transforming growth factor (TGF)-β3, TGF-β1 and bone morphogenetic protein-2 (BMP-2) in the development of this neoplastic disease.@*METHODS@#Patients in Peking University School and Hospital of Stomatology from January 2011 to February 2020 were selected, who had complete medical records, TMJSC was verified histologically after operation. The expressions of COMP, TGF-β3, TGF-β1 and BMP-2 in the TMJSC of the temporomandibular joint were detected by immunohistochemistry and quantitative real-time PCR (RT-PCR) at the protein level and mRNA level respectively, compared with the normal synovial tissue of temporomandibular joint. The histological morphology, protein expression and distribution of TMJSC tissues were observed microscopically, and the positive staining proteins were counted and scored. SPSS 22.0 statistical software was used to analyze the expression differences between the related proteins in TMJSC tissue and the normal synovial tissue of temporomandibular joint and to compare their differences. P < 0.05 indicated that the difference was statistically significant.@*RESULTS@#Immunohistochemical results showed that the positive expression of COMP in TMJSC tissues was mostly found in synovial tissues and chondrocytes adjacent to synovial tissues, and the difference was statistically significant, compared with the normal temporomandibular joint synovial tissues. The positive expression of COMP was significantly different between recurrent TMJSC and non-recurrent ones. The positive expressions of TGF-β3, TGF-β1 and BMP-2 were higher than the normal synovial tissue, and were also mostly found in the synovial cells and adjacent chondrocytes, which was further confirmed by Western blot. According to the RT-PCR results, the expressions of COMP, TGF-β3, TGF-β1 and BMP-2 in TMJSC were higher than those in the normal synovial tissue.@*CONCLUSION@#The expression of COMP in TMJSC of temporomandibular joint increased significantly, compared with the normal synovial tissue. There may be interactions between COMP and cytokines related to the proliferation and differentiation, like TGF-β3, TGF-β1 and BMP-2, which may play a potential role in the pathogenesis of TMJSC.

Association between ADAMTS14 gene polymorphism and the temporomandibular joint osteoarthritis in Chinese Han females / 北京大学学报(医学版)

OBJECTIVE@#To investigate the association between single nucleotide polymorphisms (SNP) of ADAMTS14 gene rs4747096 and osteoarthritis of the temporomandibular joint in Chinese Han females.@*METHODS@#As a case-control study, a total of 213 Chinese Han females were involved in the present study, which contained 103 temporomandibular joint osteoarthritis patients and 110 healthy people who had no symptoms or signs of temporomandibular joint osteoarthritis as control. Peripheral blood samples were collected from each participant. Genomic DNAs of temporomandibular joint osteoarthritis patients and healthy control were extracted from peripheral venous blood, which were stored in -80 °C refrigerator by using DNA extraction kits. The designed primers were used for polymerase chain reaction (PCR) amplification of specific DNA fragments. Genotype was determined by sequencing the PCR products. The software Chromas 2.22 was used to analyze the genotype. The genotype distributions, allele frequencies and genetic models between the patients and controls were compared. The age distribution was checked by t-test. Genotype and allele frequency were detected by Chi-square test.@*RESULTS@#In the present study, there were no significant differences between the osteoarthritis patients and healthy controls in terms of age. The genotype distribution was in accordance with Hardy-Weinberg equilibrium in the two groups. The genotype frequency of the ADAMTS14 (rs4747096) in the experimental group was 38.8% (AA), 55.4% (AG), and 5.8% (GG), respectively. The genotype frequency in the control group was 40.9% (AA), 43.6% (AG), and 15.5% (GG), respectively. The difference of genotype frequency of the ADAMTS14 (rs4747096) was significant between the experimental group and the control group (P=0.047). There was no significant difference in allele frequency between the two groups (P=0.415). AA and AG genotypes significantly increased the risk of the disease compared with GG in dominant model (OR=1.114, 95% CI: 1.015-1.223, P=0.028).@*CONCLUSION@#A significant correlationship was found between the ADAMTS14 (rs4747096) SNP and the temporomandibular joint osteoarthritis in Chinese Han females. The distribution of rs4747096 may be different between temporomandibular joint osteoarthritis and healthy population.

A preliminary study of murine hemangioma model by delivery of recombinant adeno-associated virus mediated human vascular endothelial growth factor-121 gene / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the feasibility of establishing a murine hemangioma model with injection of recombinant adeo-associated virus mediated human vascular endothelial growth factor-121 (rAAV-hVEGF(121)) gene.</p><p><b>METHODS</b>rAAV-hVEGF(121) was constructed, identified and then implanted to the left back ear of each mouse (1.0 x 10(11)VG in 50 microl per mouse and 10 nude mice received the injection), the rights served as controls with an injection of the same volume of phosphate buffered solution (PBS). The skin color and swelling of left back ear were observed every other day. Histological examination was carried out after mice were sacrificed 2, 4, 6, 8, 12 weeks after injection.</p><p><b>RESULTS</b>The rAAV-hVEGF(121) was correctly constructed and confirmed by restriction endonuclease analysis, polymerase chain reaction and DNA sequencing analysis. The skin of left back ear became red 2 weeks after injection and gradually exhibited a red lump which was at its utmost 12 weeks after injection. Such phenomena were not observed in right back ear. Histological examinations showed aggregates of endothelial cells by 2 weeks and at 8 weeks the swollen tissue contained many cysts filled with a mass of red cells. CD-34 staining suggested most of the newly-formed cells were endothelial cells.</p><p><b>CONCLUSIONS</b>A hemangioma model was established in mice with injection of recombinant rAAV-hVEGF(121) gene.</p>

Investigation on disc position of the temporomandibular joint in asymptomatic volunteers by magnetic resonance imaging / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the types of disc position of the temporomandibular joint (TMJ) and their relationship with age and gender in asymptomatic volunteers by magnetic resonance imaging (MRI).</p><p><b>METHODS</b>A total of 100 asymptomatic volunteers were divided into 5 groups (11-, 21-, 31-, 41-, 51 - 60 years old), 10 male and 10 female were included in each group. A total of 200 TMJs were scanned with Siemens Trio Tim 3.0T MRI system oblique-sagittally at closed- and open-mouth positions. Visual evaluation of MR images were performed on 3 neighbour slices of each TMJ (lateral, central, medial).</p><p><b>RESULTS</b>There was a normal disc-condyle relationship in 59 of the 100 asymptomatic subjects. The normal disc position, anterior disc displacement and insidious anterior disc displacement (IADD) were observed in 140 (70.0%), 14(7.0%) and 46(23.0%)joints respectively, which showed no significant differences among different age groups or between genders (P > 0.05). The maximum mouth opening of the 100 asymptomatic volunteers was (46.3 +/- 5.5) mm, with no significant difference among the various types of disc position (P > 0.05).</p><p><b>CONCLUSIONS</b>Disc displacement does exist in the asymptomatic volunteers, however, its occurrence is not associated with age and gender. IADD is the main type of the disc displacement in asymptomatic volunteers. There is no close relationship between the imaging manifestation of the disc displacement and clinical symptoms.</p>

Aquaporin-1 and aquaporin-3 expressions in the temporo-mandibular joint condylar cartilage after an experimentally induced osteoarthritis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Over 70% of the total tissue weight in the cartilage matrix consists of water, and the early-stage osteoarthritic cartilage is characterized by swelling. Water transport in the cartilage matrix and across the membranes of chondrocytes may be important in normal and pathological conditions of cartilage. The purpose of this study was to identify aquaporin-1 (AQP1) and aquaporin-3 (AQP3) expressions in the mandibular condylar cartilage after experimentally induced osteoarthritis (OA) in rats.</p><p><b>METHODS</b>An experimental temporomandibular joint OA was induced by partial discectomy in rats. The pathological characteristics of the normal, early-stage, and late-stage osteoarthritic TMJ cartilages were verified by histological techniques. The AQP1 and AQP3 gene expressions in the normal and osteoarthritic cartilages were measured using quantitative real-time reverse-transcription PCR analysis. The cartilage sections were incubated in primary polyclonal antibodies to AQP3; immunofluorescent microscopy was used to examine the AQP3 expression shown by its protein level.</p><p><b>RESULTS</b>The mRNA expression levels of AQP1 and AQP3, analyzed using quantitative PCR, revealed that AQP3 mRNA was highly up-regulated in the OA cartilage, which was considered significant. There was no notable difference in the expression of AQP1 mRNA between OA and normal controls. With the progressing of the OA, the localization of the AQP3 protein was quite different from that of the normal cartilage. Compared to the normal cartilage, the expressions of AQP3 protein were observed mainly in the proliferative zone and the upper mid-zone chondrocytes at the early-stage of OA, and were observed to appear frequently throughout the mid- and deep zone during the late-stage of OA.</p><p><b>CONCLUSIONS</b>The high expression of AQP3 mRNA in the OA cartilage and the different localization of the AQP3 protein suggest that it may play a particular role in OA pathogenesis. Further study of AQP3 function may provide new insight into the understanding of the molecular mechanisms underlying OA.</p>