OBJECTIVE: To investigate whether Salvianolic acid A (SAA) can restore cartilage endplate cell degeneration of intervertebral discs and to identify the mechanism via regulation of micro-RNA. METHODS: Cartilage endplate cells were isolated from lumbar intervertebral disc surgical samples and were treated with serum containing a series of concentrations of SAA (2, 5, and 10 ?M) for 24, 48, and 72 h to identify a proper dose and treatment time of SAA. The effect SAA on interlenkin-1β (IL-1β)-induced extracellular matrix degradation of cartilage endplate cells were analyzed by Alcian blue staining and assessment of the expression levels of ADAMTS-5, MMP3 and Col2a1. Further, the potential target miRNAs were preliminarily screened by micro-RNA sequencing combining qRT-PCR and Western blot, and then, the miRNAs mimics and inhibitors were used to verify the regulatory effect of SAA on potential target miRNAs. RESULTS: The 10 μM SAA treatment for 48 h significantly enhanced the viability of cartilage endplate cells, and increased Col2a1 expression and glycosaminoglycan accumulation that were repressed by IL-1β, and reduced the effect of IL-1β on ADAMTS-5, and MMP3. Screening analysis based on micro-RNA sequencing and Venny analysis identified the downstream micro-RNAs, including miR-940 and miR-576-5p. Then, the miR-940-mimic or miR-576-5p-mimic were transfected into CEPCs. Compared with the SAA group, the expression of ADAMTS-5 and MMP3 increased significantly and the expression of COL2A1 obviously decreased after overexpression of miR-940 or miR-576-5p in CEPCs. CONCLUSION Salvianolic acid A attenuated the IL-1β-induced extracellular matrix degradation of cartilage endplate cells by targeting regulate the miR-940 and the miR-576-5p.
Humans ; Apoptosis ; Cartilage/metabolism* ; Chondrocytes/metabolism* ; Interleukin-1beta/metabolism* ; Matrix Metalloproteinase 3/metabolism* ; MicroRNAs/metabolism*

OBJECTIVETo evaluate the effects of treadmill running exercise of different intensity on early repair of full-thickness defects on the patellofemoral articular surface and the changes in the expressions of matrix metalloproteinase-3 (MMP-3) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in SD rats.

METHODSTwenty-four male SD rats with full-thickness defects on the patellofemoral articular surface were randomly assigned into sedentary control (SED) group and low-, moderate- and high-intensity running groups (LIR, MIR, and HIR groups, respectively). The running groups were trained on treadmill for 6 consecutive weeks. Blood samples were collected to detect serum MMP-3 and TIMP-1 levels using ELISA before and after the experiment, and the femoral trochlea were collected to assess tissue repair by gross appearance scoring and O Driscoll histological scoring with Safranine O-Fast Green staining and Toluidine blue staining.

RESULTSIn rats in SED group, the defect was filled with hyaline articular cartilage-like tissues, as compared to fibrous tissues in LIR and MIR groups and subchondral bone damage in HIR group. The SED group scored the highest and HIR group the lowest among the 4 groups in gross appearance scoring and O Driscoll histological scoring. No significant differences were found in MMP-3 or TIMP-1 levels among the groups before training (P>0.05), but after 6 weeks of training, serum MMP-3 and TIMP-1 levels differed significantly among the 4 groups (P<0.05), and all the 3 running groups had a significantly higher MMP-3 level than the control group (P<0.05). After the 6-week training, TIMP-1/MMP-3 ratio was significantly higher in SED group than in the 3 running groups, and was the lowest in HIR group.

CONCLUSIONBoth low- and moderate-intensity exercise failed to promote resurfacing of full-thickness cartilage defects on the patellofemoral articular surface in rats, and high-intensity exercise even induces subchondral bone damage. The expression of MMP-3 and TIMP-1 is related to exercise, and the TIMP-1/MMP-3 ratio reflects the extent of tissue repair.

Animals ; Cartilage, Articular ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 1 ; metabolism ; Matrix Metalloproteinase 3 ; metabolism ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Wound Healing

Animals ; Collagenases ; metabolism ; Dental Bonding ; Dental Caries ; enzymology ; Dentin ; enzymology ; pathology ; Gelatinases ; metabolism ; Humans ; Matrix Metalloproteinase 14 ; metabolism ; Matrix Metalloproteinase 20 ; metabolism ; Matrix Metalloproteinase 3 ; metabolism ; Matrix Metalloproteinases ; metabolism ; Sclerosis

OBJECTIVETo study possible correlation between matrix metalloproteinases (MMPs) activities and myocardial injury after asphyxia in neonatal Wistar rats.

METHODSixty neonatal Wistar rats (7 to 10 days old) were randomly divided into four groups: control group (group D); asphyxia groups A, B and C (1 day, 7 days, 14 days after asphyxia), every group had 15 rats. In the asphyxia groups, animal model was produced by normobaric asphyxia. Groups A, B and C were sacrificed on days 1, 7 and 14 days after asphyxia, and group D rats were sacrificed on the 7 th day. Then the heart blood was taken to tested the serum cTnI. The myocardial MMPs-3 and 9 activity was measured by using immunohistochemical assay. Histological sections of the hearts were stained with hematoxylin-eosin and myocardial histopathological scores were determined under an optical microscope. The amount of myocardial collagen was measured by means of chloramines T.

RESULTScTnI was significantly higher in group A (0.3680 +/- 0.40 ng/ml) than group D (0.0783 +/- 0.06 ng/ml) (P < 0.05), and was lower in group B (0.1889 +/- 0.15 ng/ml) but still significantly different from that of group D (P < 0.05), and declined to the normal level in group C (0.1338 +/- 0.07 ng/ml), but the difference between groups C and D was not significant (P > 0.05). Myocardial tissue MMPs-3 activity was transiently high in group A (0.1847 +/- 0.04), higher in group B (0.2780 +/- 0.05) as compared to group D (0.1213 +/- 0.03) (P < 0.05 for all). The activity of MMPs-3 increased earlier than that of MMPs-9. The amount of myocardial collagen of group B (38.94 +/- 0.67) and C (40.69 +/- 0.75) was significantly greater than that of group D (P < 0.05). Myoardial tissue MMPs-3 and MMPs-9 positively correlated with myocardial histopathological scores (r = 0.669, 0.667, P < 0.05) and myocardial collagen (r = 0.482, 0.679, P < 0.05).

CONCLUSIONSIn rats with asphyxia, there was an excess activation of myocardial MMPs-3 and MMPs-9 activities and secondary to which, the quantity of myocardial collagen increased. The injuries of myocardium may be closely associated with myocardial tissue MMPs. MMPs may be used to evaluate the severity of myocardial interstitial damage.

Animals ; Asphyxia ; metabolism ; pathology ; Collagen ; metabolism ; Disease Models, Animal ; Matrix Metalloproteinase 3 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Wistar ; Troponin I ; blood

OBJECTIVE: To test the expression of NF-kappaB/p65, MMP-3 and MMP-9 in nasopharyngeal carcinoma (NPC), and analyze their relationship and the clinical significance. METHOD: All samples were examined for the expression of NF-kappaB/p65, MMP-3 and MMP-9 by streptavidin-peroxidase immunohistochemistry. Chi2-test and Spearman-test were used to exam the correlation within them and the expression difference between nasopharyngeal carcinoma and normal nasopharyngeal mucosa. RESULT: Positive expression rate of NF-kappaB/p65 in nasopharyngeal carcinoma was 78.0%; positive expressions rate of MMP-3 and MMP-9 in nasopharyngeal carcinoma were 75.6%. There was positive relationship between NF-kappaB/p65, MMP-3, MMP-9 and N stage. The expression of NF-kappaB/p65 showed a significant positive correlation with the expression of MMP-3 and MMP-9. MMP-3 also had a significant positive correlation with MMP-9. CONCLUSION The result suggests that NF-kappaB/p65, MMP-3 and MMP-9 are highly expressed in nasopharyngeal carcinoma cells. Their expression is correlated with lymph nodes metastasis, but do not have relationship with sex, age, pathological classification and clinical stage. Each of them has positive correlation with the others.
Adult ; Aged ; Female ; Humans ; Male ; Matrix Metalloproteinase 3 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Middle Aged ; NF-kappa B ; metabolism ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Transcription Factor RelA ; metabolism

OBJECTIVETo explore the curative mechanism of acupuncture treatment on osteoarthritis (OA).

METHODSForty cases of female SD rats were randomly divided into a normal group, a model group, an acupuncture group and a medication group, 10 cases in each group. OA animal model was established by using the method of heel tendon resection for unilateral hind limb. The acupuncture group was treated with electroacupuncture at "Xiqian"(ST 35) and "Housanli"(ST 36), and the medication group with inunction of Diclofenac cream, and the normal group and the model group without any treatment. The expression of matrix metalloproteinase-1, 3 (MMP-1, MMP-3) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the cartilage were observed by immunohistochemistry.

RESULTSThere were significant differences among four groups. The expressions of MMP-1, MMP-3 and TIMP-1 in the model, acupuncture and medication groups were all significantly stronger than those in the normal group (all P < 0.01). The expressions of MMP-1 and MMP-3 in the acupuncture and medication groups were down regulated and TIMP-1 expression up-regulated with significant differences as compared with the model group (all P < 0.01), and the expressions of MMP-1 and MMP-3 in acupuncture group were significantly lower, while TIMP-1 expression significantly higher than that in the medication group (all P < 0.01).

CONCLUSIONAcupuncture can down-regulate the expression of MMP-1 and MMP-3 and up-regulate the expression of TIMP1, which is superior to that of Diclofenac cream, showing that acupuncture has a certain protective effect on cartilage from OA.

Acupuncture Therapy ; Animals ; Cartilage ; chemistry ; Female ; Immunohistochemistry ; Matrix Metalloproteinase 1 ; analysis ; Matrix Metalloproteinase 3 ; analysis ; Osteoarthritis, Knee ; metabolism ; therapy ; Random Allocation ; Rats ; Tissue Inhibitor of Metalloproteinase-1 ; analysis

OBJECTIVETo study the effects of Tongxinluo on the expressions of matrix metalloproteinase-3 (MMP-3), MMP-9 and peroxisome proliferators-activated receptor gamma (PPARgamma) in atherosclerotic rabbits and explore the mechanism of its anti-atherosclerotic effect.

METHODSTwenty-four rabbits were randomized equally into control group, atherosclerotic model group (fed with high-fat diet for 14 weeks) and Tongxinluo group. The expressions of MMP-3, 9 and PPARgamma in the 3 groups were observed by means of immunohistochemistry.

RESULTSThe expressions of MMP-3, 9 and PPARgamma in the model group and Tongxinluo group were significantly higher than those in the control group (P<0.01). After high-fat diet feeding for 14 weeks, Tongxinluo group showed significantly lower expressions of MMP-3 and 9 but higher expression of PPARgamma than the model group (P<0.01).

CONCLUSIONTongxinluo can inhibit the expression of MMP-3 and 9 and increase the expression of PPARgamma, which might be the mechanism of its anti-atherosclerotic effect.

Animals ; Atherosclerosis ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Male ; Matrix Metalloproteinase 3 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; PPAR gamma ; genetics ; metabolism ; Rabbits ; Random Allocation

OBJECTIVESTo investigate the function and possible mechanisms of PIAS3 expression on the invasion of TJ905 cells.

METHODSPIAS3 overexpression vectors were constructed and PIAS3 siRNA were chemically synthesized, which were separately transfected into TJ905 cells for upregulation or downregulation of PIAS3 expression levels in TJ905 cells. After that, the invasive effects of TJ905 cells were measured by Transwell assay, and the expression of PIAS3, tissue inhibitor of metalloproteinases (TIMP)3, matrix metalloprotease (MMP)-2, and MMP-9 were identified by Western blot.

RESULTSIn vitro transfection efficiency of plasmids and oligonucleotides were separately 85.3% ± 3.1% and 95.1% ± 2.9%. PIAS3 overexpression plasmid transfection in vitro could effectively improve the expression of PIAS3 protein in TJ905 cells and inhibit the invasion of TJ905 cells (P < 0.05), and cell penetration ratio reduced from 87.9% ± 9.3% to 37.3% ± 7.9% compared with control group, while it upregulated TIMP3 and downregulated MMP-2, MMP-9 protein expression (P < 0.05); PIAS3 siRNA transfection could inhibit the PIAS3 protein expression of TJ905 cells and promote the invasion of TJ905 cells (P < 0.05), and cell penetration ratio increased from 83.9% ± 7.1% to 93.2% ± 3.1% compared with control group, while it downregulated TIMP3 and upregulated MMP-2, MMP-9 protein expression (P < 0.05).

CONCLUSIONPIAS3 expression is closely related to the invasion properties of glioma TJ905 cells.

Cell Line, Tumor ; Genetic Vectors ; Glioma ; metabolism ; pathology ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Molecular Chaperones ; genetics ; metabolism ; Neoplasm Invasiveness ; Protein Inhibitors of Activated STAT ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Tissue Inhibitor of Metalloproteinase-3 ; metabolism ; Transfection

BACKGROUNDReceptor interacting protein 1 (RIP1), which plays a key role in apoptosis, cell survival and programmed cell necrosis, is one of the most important proteins in the RIP family. The purpose of this study was to investigate the roles of RIP1 in the apoptosis, the generation of reactive oxygen species (ROS) and the expression of matrix metalloproteinases (MMPs) induced by ultraviolet B (UVB) in fibroblasts.

METHODSsiRNA targeting RIP1 was used to silence RIP1 expression in the NIH3T3 fibroblasts. The mRNA and protein levels of MMP-1 and MMP-3, caspase-3 and -8 activities, and ROS activities were determined by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR), immunoblotting, caspase activity assay, immunofluorescence, and flow cytometry.

RESULTSThe mRNA and protein expressions of MMP-1 and MMP-3 were significantly increased in RIP1 deficient NIH3T3 cells at 24 hours after UVB treatment. At 24 hours after exposure to UVB, RIP1 deficient NIH3T3 cells presented apoptotic morphology, and the apoptosis rate was significantly increased accompanied by pronounced increase in caspase-8 and -3 activities. ROS production was inhibited by UVB at 12 hours in RIP1 deficient NIH3T3 cells.

CONCLUSIONRIP1 is involved in NIH3T3 cell damage induced by UVB via participating in the apoptosis, expression of MMPs and ROS production.

Animals ; Apoptosis ; GTPase-Activating Proteins ; genetics ; metabolism ; Matrix Metalloproteinase 1 ; genetics ; metabolism ; Matrix Metalloproteinase 3 ; genetics ; metabolism ; Matrix Metalloproteinases ; genetics ; metabolism ; Mice ; NIH 3T3 Cells ; RNA, Small Interfering ; Reactive Oxygen Species ; metabolism ; Ultraviolet Rays

Keloids are pathologic proliferations of the dermal layer of the skin resulting from excessive collagen production and deposition. Hepatocyte growth factor (HGF) increases the expression of matrix metalloproteinase (MMP)-1 and suppresses collagen synthesis to modulate extracellular matrix turnover. To investigate the anti-fibrotic effects of HGF, we examine the mRNA expression of collagen types I and III and matrix metalloproteinase (MMP-1, MMP-3) on human dermal fibroblast (HDF) cell lines and keloid fibroblasts (KFs, n = 5) after adding various amount of HGF protein. We also evaluated the enzymatic activity of MMP-2, MMP-9 by zymograghy. In HDFs treated with TGF-beta1 and HGF protein simultaneously, both type I and III collagen mRNA expression significantly decreased (P < 0.05). Expression of MMP-1, MMP-3 mRNA also decreased. However, the mRNA expression of MMP-1, MMP-3 significantly increased in KFs with increasing amount of HGF in dose dependent manner (P < 0.05). The enzymatic activities of MMP-2 increased with increasing HGF protein in a dose-dependent manner. However, the enzymatic activity of MMP-9 did not change. These results suggest that the anti-fibrotic effects of HGF may have therapeutic effects on keloids by reversing pathologic fibrosis.
Cells, Cultured ; Collagen Type I/genetics/metabolism ; Collagen Type III/genetics/metabolism ; Fibroblasts/drug effects/metabolism ; Hepatocyte Growth Factor/*pharmacology ; Humans ; Keloid/*metabolism/pathology ; Matrix Metalloproteinase 1/genetics/metabolism ; Matrix Metalloproteinase 2/metabolism ; Matrix Metalloproteinase 3/genetics/metabolism ; Matrix Metalloproteinase 9/metabolism ; RNA, Messenger/metabolism ; Transforming Growth Factor beta1/pharmacology