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 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

OBJECTIVES: To determine the level of mRNA expression of various members of the matrix metalloproteinase and tissue inhibitors in uterine leiomyoma compared with unaffected myometrium. Materials & Method: 30 cases of portions of leiomyoma and myometrium were collected immediately followimg hysterectomy. Thirteen cases were from proliferative phase and seventeen were from secretory phase of menstrual cycle. The mean age was 43.7years old. The level of expression of mRNAs of interstitial collagenase, gelatinase, stromelysin, TIMP-1,-2,-3 was determined by reverse transcriptase-polymerase chain reaction(RT-PCR) and normalized to GAPDH(glyceraldehyde-3-phosphate dehydrogenase) mRNA. RESULTS: Myometrium and leiomyoma expressed all the members of above mentioned matrix metalloproteinase family and tissue inhibitors. Leiomyoma expressed a significantly higher level of stromelysin-3 during secretory phase, an extremely lower level of 92kDa gelatinase and a significantly lower level of TIMP-3. The immunohistochemical localization of TIMP-3 was smooth muscle cell and arteriole wall of myometrium and leiomyoma. CONCLUSIONS: The increased expression of stromelysin-3 in uterine leiomyoma compared with myometrium suggests that this MMP may be involved in the formation of a more fibrous extracellular matrix in leiomyoma. The extremely lower expression of 92kDa gelatinase of leiomyoma means that leiomyoma do not invade myometrium and forms a separated mass. Decreased expression of TIMP-3 of leiomyoma suggests that TIMP-3 is required for differentiation and homeostasis of extracellular matrix of normal myometrium and function as a suppressive role of tumor development
Animals ; Arterioles ; Extracellular Matrix ; Female ; Gelatinases ; Homeostasis ; Humans ; Hysterectomy ; Leiomyoma* ; Matrix Metalloproteinase 1 ; Matrix Metalloproteinase 3 ; Menstrual Cycle ; Mice ; Myocytes, Smooth Muscle ; Myometrium* ; RNA, Messenger ; Tissue Inhibitor of Metalloproteinase-3

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

PURPOSE: To investigate subcellular localizations of wild type myocilin and trunctated (Q368X) myocilin in cultured human trabecular meshwork (TM) cells. METHODS: GFP and GFP tagged truncated myocilin, full-length myocilin, and stromelysin were expressed in TM cells using adenoviral vectors, and their secretory properties were examined by western blotting. To determine the subcellular localizations of myocilins, cellular organelles of the infected TM cells were stained with antibodies or orgenelle specific fluorescent indicators and examined under confocal microscope. RESULTS: Wild type myocilin was expressed as discrete fine vesicles in the perinuclear region of TM cells as well as in ER, but not in microtubules or mitochondria. Colocalization of wild type and truncated myocilin, indicative of in vivo interaction of the proteins, was also observed in cells co-expressing the proteins. Truncated myocilin was found to be accumulated as aggregates in ER, and inhibited the secretion of normal myocilin. CONCLUSIONS: Our result suggests that intracellular accumulation of truncated myocilin may cause a dysfunction of the cells, resulting in alterations in structural compartmentalization of trabecular ECM and obstruction of aqueous outflow.
Antibodies ; Blotting, Western ; Humans ; Matrix Metalloproteinase 3 ; Microtubules ; Mitochondria ; Organelles ; Trabecular Meshwork*

OBJECTIVE: To investigate the expression level of serum matrix metalloproteinase 3 (MMP3) in early rheumatoid arthritis (ERA) patients with normal C-reaction protein (CRP) or erythrocyte sedimentation rate (ESR), and the significance in disease assessment. METHODS: In the study, 133 cases of early RA patients, 25 osteoarthritis (OA) patients and 60 healthy controls in Peking University People's Hospital from 2011 to 2015 were included. The RA patients were further divided into 4 groups according to levels of CRP and ESR: 88 patients with increased CRP and increased ESR, 15 patients with normal CRP and normal ESR, 17 patients with normal CRP but increased ESR, and 13 patients with increased CRP but normal ESR. All the clinical information of the patients was collected, and the serum MMP3 levels of both patients and healthy controls were detected by enzyme-linked immune sorbent assay (ELISA). RESULTS: The serum MMP3 level of RA patients with normal CRP and/or normal ESR [(72.89±6.34) μg/L] was obviously higher than that of OA patients [(42.87±4.14) μg/L] (P=0.002) and healthy controls [(31.62±2.88) μg/L] (P<0.001). The serum MMP3 levels of the patients with normal CRP and normal ESR [(47.04±9.64) μg/L] were higher than those of the healthy controls, and there was statistical significance between the two groups (P<0.05). The serum MMP3 levels of the patients with increased CRP but normal ESR [(94.18±9.11) μg/L] and the patients with normal CRP but increased ESR [(79.42±10.60) μg/L] were both higher than those of the OA patients and healthy controls, and there was obvious statistical difference (P<0.05). In the early RA patients with normal CRP and/or normal ESR, the serum MMP3 level was positively correlated with the CRP level (r=0.336, P=0.024). The positive rate of MMP3 in the patients with normal CRP and/or normal ESR was 44.44%, higher than the positive rate of CRP (28.89%) and the positive rate of ESR (37.78%). In these early RA patients, the positive rate was 52.94% in the patients with normal CRP but increased ESR and 53.85% in the patients with increased CRP but normal ESR. CONCLUSION The detection of the serum MMP3 level was significant in the assessment of early RA patients within 2-year duration who had normal CRP or ESR value.
Arthritis, Rheumatoid/diagnosis* ; Blood Sedimentation ; Humans ; Matrix Metalloproteinase 3/blood* ; Osteoarthritis

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 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

OBJECTIVETo investigate the relationship between the genetic polymorphisms of matrix metalloproteinase-2 (MMP-2) (-735) and matrix metalloproteinase-3 (MMP-3) (-1171) and the susceptibility to silicosis.

METHODSA case-control study was conducted in 113 patients diagnosed with stage I silicosis (case group) and 115 dust-exposed workers without silicosis (control group); the two groups had the same sex, ethnic group, and type of dust and similar age and cumulative exposure time. Polymerase chain reaction-restriction fragment length polymorphism was used to determine the genotypes of MMP-2 (-735) and MMP-3 (-1171).

RESULTSNo significant difference was observed in age, cumulative exposure time, or smoking rate between cases and controls (P > 0.05). The frequencies of genotypes C/C, C/T, and T/T at MMP-2 C-735T in the case group were 57.5% (65/113), 31.0% (35/113), and 11.5% (13/113), respectively, which were significantly different from those of the control group (69.6% (80/115), 26.9% (31/115), and 3.5% (4/115)), χ² = 6.542, P < 0.05). The frequencies of T allele in cases and controls were 27.0% and 17.0%, respectively, which were significantly different from each other χ² = 6.704, P < 0.05). Carriage of T allele at MMP-2 C-735T increased the risk of silicosis (OR = 1.811, 95% CI: 1.151-2.847). The frequencies of genotypes 6A/6A, 5A/6A, and 5A/5A at MMP-3 A-1171A were 67.2% (76/113), 24.8% (28/113), and 8.0% (9/113), respectively, in the case group, versus 59.1% (68/115), 37.4% (43/115), and 3.5% (4/115) in the control group (χ² = 5.519, P > 0.05).

CONCLUSIONGenetic polymorphism at MMP-2 C-735T is significantly associated with the development of silicosis. Carriage of T allele increases the risk of silicosis among workers exposed to dust. No significant association was found between MMP-3 A-1171A polymorphism and silicosis in this study.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Genetic Predisposition to Disease ; Genotype ; Humans ; Matrix Metalloproteinase 2 ; genetics ; Matrix Metalloproteinase 3 ; genetics ; Middle Aged ; Polymorphism, Genetic ; Silicosis ; genetics

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