Association between -1607 1g/2g polymorphism of mmp1 and temporomandibular joint anterior disc displacement with reduction

Abstract Anterior disc displacement with reduction (DDWR) is considered one of the most common disorders within the temporomandibular joint (TMJ), with a prevalence of 41% in adults. Matrix metalloproteinases play an important role in the degradation of the TMJ and the matrix metalloproteinase 1 (MMP1) 1607 1G/2G polymorphism increases the local expression of MMP1 thus leading to accelerated degradation of the extracellular matrix. The objective of this study was to evaluate the association between the 1607 1G/2G polymorphism of MMP1 gene and DDWR in a group of Mexican individuals from western Mexico. A total of 67 unrelated individuals, between the ages of 18 and 36 years, of both genders, were included in this study. Study participants with DDWR were required to meet the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD), while a second control group of 90 individuals without DDWR were also included. Both groups were required to have paternal and maternal ancestry (grandparents) of the same geographic and ethnic region. Genotypes were determined using the nested PCR technique. The 1G/2G polymorphism was found in 68.7%, followed by 2G/2G in 25.4% and 1G/1G in 6.0% of the cases group. While the prevalence in the control group was 55.5% for the 1G/2G polymorphism, 26.6% for 1G/1G and 17.7% for 2G/2G. An association was found between the 2G allele of the 1607 1G/2G polymorphism of MMP1 gene and the presence of DDWR in the patients of western Mexico.

Resumo O deslocamento anterior do disco com redução (DADR) é considerado um dos distúrbios mais comuns na articulação temporomandibular (ATM), com prevalência de 41% em adultos. As metaloproteinases da matriz desempenham um papel importante na degradação da ATM e o polimorfismo 1607 1G/2G da metaloproteinase da matriz 1 (MMP1) aumenta a expressão local da MMP1, levando à degradação acelerada da matriz extracelular. O objetivo deste estudo foi avaliar a associação entre o polimorfismo 1607 1G/2G do gene MMP1 e a DADR em um grupo de indivíduos mexicanos do oeste do México. Um total de 67 indivíduos não relacionados, com idades entre 18 e 36 anos, de ambos os sexos, foram incluídos neste estudo. Os participantes do estudo com DADR foram obrigados a cumprir os Critérios de Diagnóstico de Pesquisa para Disfunções Temporomandibulares (CDP/DTM), enquanto um segundo grupo controle de 90 indivíduos sem DADR também foi incluído. Ambos os grupos tinham ascendência paterna e materna (avós) da mesma região geográfica e étnica. Os genótipos foram determinados pela técnica de nested PCR. o polimorfismo 1G/2G foi encontrado em 68,7%, seguido por 2G/2G em 25,4% e 1G/1G em 6,0% do grupo de casos. Enquanto a prevalência no grupo controle foi de 55,5% para o polimorfismo 1G/2G, 26,6% para 1G/1G e 17,7% para 2G/2G. Foi encontrada uma associação entre o alelo 2G do polimorfismo 1607 1G/2G do gene MMP1 e a presença de DADR nos pacientes do oeste do México.

Changes of fibrotic factors in the recovery process of skeletal muscle strain / 中国组织工程研究

BACKGROUND: Skeletal muscle injury is the most common sports injury, which can develop fibrosis and then impair muscle function. Varieties of fibrotic factors are involved in the repair of skeletal muscle and effectively regulate the process of tissue fibrosis. OBJECTIVE: To establish a rat model of acute gastrocnemius strain, observe the changes of fibrotic factors in the recovery of injury and explore the possible roles of fibrotic factors. METHODS: Totally 112 male Sprague-Dawley rats were randomly divided into control group (n=56) and injury group (n=56). The left gastrocnemius muscle was injured by a rat gastrocnemius pull-off device in the injury group. Each group was further subdivided into immediately group, 2 days group, 4 days group, 7 days group, 14 days group, 21 days group and 28 days group depending on the sampling time points. There were eight rats in each subgroup. The control group did not receive any intervention. The left gastrocnemius muscle of the injury group was strained by specialized equipment. Muscle fiber injury and inflammation were observed by hematoxylin-eosin staining. The expression of transforming growth factor β1 (TGF-β1), connective tissue growth factor (CTGF), matrix metalloproteinase-1 (MMP-1), and matrix metalloproteinase tissue inhibitor 1 (TIMP-1) were detected by western blot. The experiment was approved by the Sports Science Experimental Ethics Committee of Beijing Sport University. RESULTS AND CONCLUSION: Hematoxylin-eosin staining results showed that the muscle fibers of the gastrocnemius muscle arranged disorderedly and inflammatory cells were aggregated in the injury group. Compared with the control group, protein expression levels of TGF-β1 and CTGF significantly increased in the injury group from the 2nd day throughout the recovery (P 0.05). TIMP-1 expression significantly increased from the 7th day until the 28th day (P < 0.05). These findings indicate that TGF-β1/CTGF signal pathway may be successively activated during the recovery of skeletal muscle strain, triggering the occurrence and development of skeletal muscle fibrosis. Meanwhile, MMP-1 and TIMP-1 protein may be activated to participate in the recovery of injury.

Preventive effect and mechanism of ultrafine powder of Dendrobium candidum on photoaging model mice / 中国中药杂志

Chinese herbal medicine ultrafine powder has become a research hotspot for the addition of cosmetic raw materials. Dendrobium candidum is a traditional Chinese herbal medicine. Its extract and stem extract are already cosmetic raw materials and its water extract has the effect of preventing photoaging,but D. candidum ultrafine powder has not been accepted as a raw material for cosmetics,and no relevant research on photoaging prevention has been reported. In this experiment,the ultra-fine powder and fine powder of D. candidum to prevent photoaging were observed and compared,and its mechanism of action was discussed to provide a basis for the prevention of skin photoaging products. Seventy-two female ICR mice were randomly divided into normal group,model group,solvent group,titanium dioxide(Ti O2) group,isooctyl salicylate(2-ES) group,D. candidum ultrafine powder 1(DP1),ultrafine powder 2(DP2) and fine powder(DP3) groups. The photoaging model was established by ultraviolet irradiation for 8 weeks,and the model was intervened while modeling. The skin wrinkle grade,elastic parameters,skin microcirculation blood flow,skin structure and pathological changes(skin thickness,skin collagen fiber,elastic fiber) were observed,the skin transforming growth factor-β1(TGF-β1),Smad3 levels were determined,and the type Ⅰ and type Ⅲ collagen,matrix metalloproteinase-1(MMP-1),activated protein-1(AP-1),VEGF expression were detected. The results showed that ultrafine powder(DP1,DP2) significantly reduced the wrinkle level and skin blood flow of the model mice(P<0. 05,P<0. 01); DP1,DP2 and DP3 could significantly reduce the thickness of the epidermis(P<0. 001),improve collagen fiber,elastic fiber hyperplasia,and distortion and decrease VEGF expression,and DP1 is better than DP2 and DP3; each group could up-regulate type Ⅰ collagen,down-regulate type Ⅲ collagen,AP-1,MMP-1 protein expression,and DP1 improvement optimal. However,it has no obvious effect on TGF-β1 and Smad3. The ultrafine powder and fine powder of D. candidum have certain preventive effect on photoaging,and the effect of ultrafine powder is better than that of fine powder. Ultrafine powder may down-regulate the expression of type Ⅲ collagen,AP-1 and MMP-1 by up-regulating type Ⅰ collagen. Inhibition of collagen degradation plays a role in preventing photoaging.

Infradiaphragmatic Stasis-Expelling Decoction reduce the expression of TIMP-1 contributing to inhibition of rat liver fibrosis / 实用医学杂志

Objective To investigate the protective effect and its mechanism of infradiaphragmatic stasis-expelling decoction on hepatic fibrosis in rats. Method One hundred and ten SD rats were divided into four groups:the normal control group ,model group ,low dose group and high dose group of infradiaphragmatic stasis-expelling decoction. In addition to the normal control group,rats in other groups were subcutaneously given pure CCl4 to set up the liver fibrosis model. Twelve weeks later,the serum liver biochemical indexes,including ALT,AST, ALB,TP and T-bil,the Ishak score about liver fibrosis ,the positive expression of TIMP-1 were compared among groups. Results Compared with the fibrotic group,the levels of serum ALT,AST and T-bil were lower in low dose group,the level of serum TP and ALB were increased in high dose group Levels of serum ALT,AST and T-bil were significantly lowered in high dose group,the levels of serum of TP and ALB were significantly increased in high dose group. Compared with model group,the Ishak score about liver fibrosis was significantly lowered(P<0.05). Numbers of cells with positive expression of TIMP-1 in liver tissue was reduced. Conclusion Infradiaphragmatic stasis-expelling decoction could inhibit the expression and activity of TIMP-1 contributing to the effect of antiliver fibrosis.

Effects of Qishen Erlian Decoction on Serum MMP-1 and TIMP-1 in Liver Fibrosis Model Rats / 中国中医药信息杂志

Objective To investigate the effects of Qishen Erlian Decoction on serum MMP-1 and TIMP-1 levels in thioacetamide (TAA) induced liver fibrosis rats; To discuss its mechanism of action. Methods Liver fibrosis model was created by the TAA gavage method. 120 SD male rats were randomly assigned to control group, model group, colchicine group, Qishen Erlian Decoction low-, medium- and high-dose group (20 in each group). Each medication group was given relevant medicine for gavage. Control group and model group were given the same amount of normal saline for gavage, once a day for 5 weeks. HE staining and Masson trichrome staining were used to observe the pathological changes in liver tissue and liver tissue damage. Biochemistry, radioimmunoassay, and ELISA were used to detect the serum liver function, hepatic fibrosis index, MMP-1 and TIMP-1 levels. Results Compared with the model group, serum ALT, AST, TBIL, γ-GGT, HA, LN, Ⅳ-C and PCⅢ levels, MMP-1 and the ratio of MMP-1/TIMP-1 increased significantly and level of TIMP-1 decreased significantly in Qishen Erlian Decoction groups, with statistical significance (P<0.05, P<0.01). And there is a certain dose-effect relationship, with Qishen Erlian Decoction high-dose group the best effect. Conclusion Qishen Erlian Decoction can improve the liver function and liver fibrosis indexes, regulate the level of MMP-1 and TIMP-1, and prevent the progression of liver fibrosis.

Wound healing activity of Ullucus tuberosus, an Andean tuber crop

Objective: This study was designed to investigate the wound healing activity of aqueous extracts of Ullucus tuberosus (U. tuberosus) using in vitro models. Methods: Lyophilized pulp and acetone extracts of U. tuberosus were produced using ultrasound extraction. The capacity for collagenase activation was evaluated using fluo-rescence detection of the enzymatic activity. Then, the influence of U. tuberosus extracts on cell proliferation, cell migration and synthesis of the extracellular matrix (ECM) proteins, metalloproteinase (MMP-1) and pro-collagen was analyzed using human dermal fibroblasts in culture. Results: An increase in collagenase activity of 12%supports the utility of U. tuberosus as an agent for scar treatment. In addition, the extracts showed an increase in the pro-liferation and migration of human dermal fibroblasts and the production of pro-collagen and MMP-1 after treatment with U. tuberosus extracts. The increase in proliferation, migration and pro-collagen levels positively influenced the regeneration of scarless tissue during the proliferation phase, whereas the increase in MMP-1 may have favored the wound healing process during the remodeling and cellular differentiation phases. Conclusion: The results of this study show for first time that U. tuberosus is a promising candidate to support scarless tissue regeneration.

Expression and significance of MMP-1 and TIMP-1 in the nucleus pulposus and anulus fibrosus of intervertebral disc with different lesions / 局解手术学杂志

Objective To explore the expression and significance of MMP-1 and TIMP-1 in the nucleus pulposus and anulus fibrosus of intervertebral disc with different lesions.Methods From August 2014 to August 2016,34 cases of lumbar disc herniation (the observation group) and 34 case of surgical removal of intervertebral disc in patients with vertebral trauma caused by sudden trauma (the control group) in our hospital were stuided.The nucleus pulposus and anulus fibrosus of intervertebral disc were separated,and the expression of MMP-1 and TIMP-1 were detected by immunohistochemistry.The relationship between MMP-1 and TIMP-1 expression and intervertebral disc degeneration was analyzed.Results The MMP-1 and TIMP-1 expression of nucleus pulposus in the observation group were significantly higher than those in the control group(P>0.05);and the MMP-1 and TIMP-1 expression of nucleus pulposus decreased successively in prominent type,extrusion type,free type lumbar intervertebral disc degeneration(P0.05);and the MMP-1 and TIMP-1 expression of anulus fibrosus decreased successively in prominent type,extrusion type,free type lumbar intervertebral disc degeneration(P<0.05).Patients with high expression of MMP-1 and TIMP-1 in nucleus pulposus and anulus fibrosus had more risk of degenerative disc disease(P<0.05).As a sensitivity prediction of intervertebral disc degeneration,the sensitivity of MMP-1 was 82.35% while the specificity was 94.11%,and the sensitivity of TIMP-1 was 79.41% while the specificity was 88.24%.The prediction sensitivity of intervertebral disc degeneration was 90.91% and the specificity was 97.14% when combined MMP-1 with TIMP-1.Conclusion The expression of MMP-1 and TIMP-1 in the nucleus pulposus and anulus fibrosus of intervertebral disc degeneration was significantly increased,which is of great clinical significance in the diagnosis of intervertebral disc degeneration.

Wound healing activity of Ullucus tuberosus, an Andean tuber crop

Objective This study was designed to investigate the wound healing activity of aqueous extracts of Ullucus tuberosus (U. tuberosus) using in vitro models. Methods Lyophilized pulp and acetone extracts of U. tuberosus were produced using ultrasound extraction. The capacity for collagenase activation was evaluated using fluorescence detection of the enzymatic activity. Then, the influence of U. tuberosus extracts on cell proliferation, cell migration and synthesis of the extracellular matrix (ECM) proteins, metalloproteinase (MMP-1) and pro-collagen was analyzed using human dermal fibroblasts in culture. Results An increase in collagenase activity of 12% supports the utility of U. tuberosus as an agent for scar treatment. In addition, the extracts showed an increase in the proliferation and migration of human dermal fibroblasts and the production of pro-collagen and MMP-1 after treatment with U. tuberosus extracts. The increase in proliferation, migration and pro-collagen levels positively influenced the regeneration of scarless tissue during the proliferation phase, whereas the increase in MMP-1 may have favored the wound healing process during the remodeling and cellular differentiation phases. Conclusion The results of this study show for first time that U. tuberosus is a promising candidate to support scarless tissue regeneration.

Effects of metronidazole and amoxicillin combination on MMP-1,MMP-8 and TIMP-1 level in gingival crevic-ular fluid of patients with aggressive periodontitis / 实用口腔医学杂志

40 cases(control group)with aggressive periodontitis (AgP)received scaling and root planning (SRP)and 38 cases(test group)received SRP followed by oral administration of amoxicillin plus metronidazole for 7 d.Gingival crevicular fluid samples were exam-ined for the levels of MMP-1,MMP-8 and tissue TIMP-1 by ELISA before therapy,3 and 6 months after therapy,TIMP-1 /MMP-1 and TIMP-1 /MMP-8 ratios were calculated.The levels of MMP-1 and MMP-8 were decreased in both groups (P <0.05)at 3 and 6 months after therapy.TIMP-1 /MMP-1 and TIMP-1 /MMP-8 ratios were increased in the 2 groups(P <0.05)after treatment,3 months after therapy the ratio in test group was higher than that in control group(P <0.05).

Adenophora remotiflora protects human skin keratinocytes against UVB-induced photo-damage by regulating antioxidative activity and MMP-1 expression

BACKGROUND/OBJECTIVES: Chronic ultraviolet (UV) exposure-induced reactive oxygen species (ROS) are commonly involved in the pathogenesis of skin damage by activating the metalloproteinases (MMP) that break down type I collagen. Adenophora remotiflora (AR) is a perennial wild plant that inhabits Korea, China, and Japan. The present study investigated the protective effects of AR against UVB-induced photo-damage in keratinocytes. MATERIALS/METHODS: An in vitro cell-free system was used to examine the scavenging activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical and nitric oxide (NO). The effect of AR on ROS formation, antioxidant enzymes, elastase, MMP-1 level, and mRNA expression of MMP-1 were determined in UVB-irradiated human keratinocyte HaCaT cells. RESULTS: AR demonstrated strong DPPH free radical and NO scavenging activity in a cell-free system exhibiting IC50 values of 1.88 mg/mL and 6.77 mg/mL, respectively. AR pretreatment dose-dependently attenuated the production of UVB-induced intracellular ROS, and antioxidant enzymes (catalase and superoxide dismutase) were enhanced in HaCaT cells. Furthermore, pretreatment of AR prevented UVB-induced elastase and collagen degradation by inhibiting the MMP-1 protein level and mRNA expression. Accordingly, AR treatment elevated collagen content in UVB-irradiated HaCaT cells. CONCLUSION: The present study provides the first evidence of AR inhibiting UVB-induced ROS production and induction of MMP-1 as a result of augmentation of antioxidative activity in HaCaT human keratinocytes. These results suggest that AR might act as an effective inhibitor of UVB-modulated signaling pathways and might serve as a photo-protective agent.

Inhibitory effect of Aralia elata ethanol extract against skin damage in UVB-exposed human keratinocytes and human dermal fibroblasts / 한국영양학회지

PURPOSE: Solar ultraviolet (UV) radiation causes inflammation and matrix metalloproteinase (MMP) overexpression and extracellular matrix depletion, leading to skin photoaging such as wrinkle formation, dryness, and sagging. Activation of MMP is influenced by various molecules such as reactive oxygen species (ROS), proinflammatory cytokines, and transient receptor potential vanilloid type (TRPV)-1, which are increased in UV-irradiated skin cells. Aralia elata (AE) ethanolic extract was reported to inhibit ROS generation caused by UVB-irradiation in keratinocytes. In this study, we investigated the photoprotective effect of AE ethanolic extract on UVB-irradiated human keratinocytes (HaCaT) and human dermal fibroblasts (HDF). METHODS: AE was freeze-dried, extracted in 70% ethanol, and concentrated. Skin cells were treated with AE extract for 24 h and then exposed to UVB (55 mJ/cm2). After 48 h of incubation, proinflammatory cytokines, MMP-1, type-1 procollagen, and TRPV-1 levels were measured by ELISA or Western blotting. RESULTS: Treatment with AE extract (100 µg/mL) significantly inhibited UVB-induced IL-6, IL-8, and PGE2 production in HaCaT by 25.6%, 5.3%, and 70.2%, respectively, and also inhibited elevation of MMP-1 and TRPV-1 caused by UVB irradiation by 20.0% and 41.9%, respectively (p < 0.05). In HDF, AE extract treatment significantly inhibited both elevation of MMP-1 and reduction of type-1 procollagen caused by UVB irradiation (p < 0.05). In addition, type-1 procollagen was elevated by AE extract treatment in normal HDFs (p < 0.05). CONCLUSION: AE 70% ethanol extract has photoprotective ability via reduction of proinflammatory mediators, TRPV-1 and MMP-1 production, and elevation of collagen synthesis. Our findings suggest that AE extract might be a good natural material to protect against UVB-induced premature skin aging.

Effects of Er-Miao-San extracts on TNF-alpha-indueed MMP-1 expression in human dermal fibroblasts

BACKGROUND: Various health benefits have been attributed to Er-Miao-San (EMS), a traditional Chinese herbal formulation that contains equal amounts of cortex phellodendri (Phellodendron amurense Ruprecht) and rhizoma atractylodis (Atractylodes lancea D.C). However, its effect on the anti-inflammatory activity in human dermal fibroblasts (HDFs) and the mechanism underlying this effect are unknown. RESULTS: This study investigated the effects of EMS on TNF-α-induced MMP-1 expression in HDFs. Our data show that EMS inhibited TNF-α-induced MMP-1 expression in a concentration-dependent manner. Interestingly, EMS maintained IkB content without inhibiting the phosphorylation of MAPKs, which are well-established upstream kinases of NF-kB. Moreover, EMS reduced the level of nuclear p65 protein in HDFs. Luciferase assay revealed that EMS inhibits the transcriptional activity of NF-kBbystabilizing IkB. Our results show that EMS exerts its anti-inflammatory effect by inhibiting NF-kB-regulated genes such as IL-1ß and IL-8. Moreover, EMS effectively inhibited TNF-α-induced expression of MMP-1 via the NF-kBpathway. CONCLUSIONS: Taken together, our data suggest that EMS could potentially be used as an anti-inflammatory and anti-aging treatment.

Effect of Jinfeng particles on MMP-1/TIMP-1 expression in articular cartilage of gouty arthritis in rabbits / 中国免疫学杂志

Objective:To explore the effect of Jinfeng particles on the expression of MMP-1/TIMP-1 in cartilage of gout arthritis rabbit′s articular,to explicit treatment mechanism of gouty arthritis. Methods:Perfusion of ethambutol and adenine,inject MSU into joint cavity of rabbits to copy the model of gouty arthritis. After successful mode,were randomly divided into model group,Jinfeng granules in high,medium and low dose group(19. 6,9. 8,4. 9 g/kg),allopurinol group(0. 028 g/kg),Tongfengding group(0. 448 g/kg) ,another blank group. Administered for 7 days. 2 h after the last administration,were taken and painted articular cartilage tissue,to observe the changes of the pathological morphology;and the use of immune cell detection of the expression of TIMP-1,MMP-1,MMP-1/TIMP-1 ratio calculation. Results: Jinfeng granule could obviously reduce the articular cartilage of rats with inflammatory cells infiltration,hyperemia, swelling and necrosis, could significantly inhibit the expression of MMP-1 ( P< 0. 05 ) , stimulate TIMP-1 activation(P< 0. 01),and significantly reduce the ratio of MMP-1/TIMP-1(P< 0. 05). Conclusion: Jinfeng particles can effectively inhibit the pathological damage of gouty arthritis,upside the expression of TIMP-1 and its mechanism may be related to the inhibition of MMP-1,regulated the balance of MMP-1/TIMP-1,and then exert anti-inflammatory effects.

The expression of MMP-1 and MMP-2 of periodontal ligament fibroblasts treated by tea polyphenols and LPS / 实用口腔医学杂志

Objective:To survey the expression of MMP-1,MMP-2 of human periodontal ligament cells(HPDLCs)treated by tea polyphenols(TP)and lipopolysaccharide(LPS).Methods:HPDLCs were in vitro cultured in vitro and treated by TP(200 μg/ml) and /or LPS(100 μg/ml)for 24,48 and 72 h respectively,the secretion of MMP-1 and MMP-2 were examined by ELISA,MMP-1 and MMP-2 mRNA expression was examined by real-time PCR.Results:The secression and mRNA expression of MMP-1 and MMP-2 of HPDLCs increased by LPS treatment and significantly inhibited by TP at the different times.Conclusion:TP can inhibit the col-lagen degradation of HPDLCs mediated by LPS.

Expression of MMP-1 and PTEN protein in the lesions of basal cell papilloma and its correlation with skin photoaging / 中国肿瘤临床

Objective:To investigate the expression of MMP-1 and PTEN protein in basal cell papilloma (BCP), as well as their correlation with skin photoaging. Methods:Immunohistochemistry technique via Elivison method was employed to measure the expres-sion of MMP-1 and PTEN protein in lesions from 50 cases of BCP on exposed areas, 50 cases on non-exposed areas, and 30 normal controls. We compared the differences among the three groups and analyzed the result. A total of 90 BCP cases on exposed areas were randomly divided into three groups. Titanium dioxide cream and placebo were respectively applied in the trial groups twice daily for 12 weeks, whereas the control group was non-administered. After 12 weeks, the MMP-1 in the lesions of the three groups was measured and compared. Results:The expression scores of MMP-1 on exposed areas were significantly higher than those in the control group (P0.05). The expression scores of PTEN protein on exposed areas and on non-exposed areas were significantly lower than that in control group (P<0.01). The expression scores of MMP-1 in the group that used titanium dioxide were evidently lower than those in control group after 12 weeks (P<0.05). Conclu-sion:MMP-1 is overexpressed in BCP on exposed areas. PTEN protein is underexpressed in BCP of exposed areas and non-exposed ar-eas. Skin photoaging is a possible cause of BCP on exposed areas.

Changes in expression of matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in pathological scar tissue of rabbit ears after scar-skin replantation / 中华医学美学美容杂志

Objective To observe the changes of MMP-1 and TIMP-1 expression in the pathological scar tissue after scar-skin replantation and to explore the mechanism of treating pathological scars with scar-skin replantation through a rabbit ear model.Methods Rabbit ears were used to establish the hypertrophic scar animal model in this study.Specimens were taken for three times:normal skin,hypertrophic scar and scar-skin replantion separately.We then performed HE staining,Masson staining and immunohistochemical staining to observe the expression of MMP-1 and TIMP-1 in these three groups of specimens.Results The expression of both MMP-1 and TIMP-1 significantly increased in the hypertrophic scar tissue after scar-skin replantation compared with the control group (P＜0.01).The expression of MMP-1 increased more significantly than that of TIMP-1 (P＜0.01).Conclusions The mechanism of scar-skin replantation's effect in the treatment for hypertrophic scar is relevant to the imbalance in the interaction between MMP-1 and TIMP-1 in the scar tissue.

Regulation of activated genes of fibroblasts and their improvement of skin aging / 中华医学美学美容杂志

Objective To investigate the possible mechanisms using different chemicals to improve the skin aging by the effect of estrogen,vitamin E,vitamin C on the transcriptional level of MMP-1 and HAS-2 mRNAs in cultured human skin fibroblast in vitro.Methods The different concentrations of estrogen,vitamin E,vitamin C were put into cultured human skin fibroblasts.After 24 hours,RT-PCR was used to detect the effects of the different chemicals on the transcriptional levels of the HAS-2 and MMP-1 mRNAs in fibroblasts.Results The effects of estrogen and vitamin C on the MMP-1 mRNA had significant difference between the high dose group and blank control group,and between the low dose group and blank control group (P＜0.05) ; the difference between the low dose group and high dose group was not statistically significant (P＞0.05).The effects of vitamin E and vitamin C on Has-2 mRNA also had significant difference between the high dose group and the blank control group,and between the low dose group and blank control group (P＜0.05),but the difference between the low dose group and high dose group was not statistically significant (P＞0.05).Conclusions Estrogen and vitamin C can reduce the transcription of MMP-1 gene,which decreases the degradation of collagen in the skin; vitamin E and vitamin C can increase the transcription of HAS-2 gene,which may possibly increase the synthesis of hyaluronic acid to alleviate the skin aging.

Curcumin Induced Decreased Expression of Type I Collagen in Human Skin Fibroblast Through Down-regulation of Smad2/3 Expressions / 대한피부과학회지

BACKGROUND: Imbalance among TGF-beta/Smad pathway, MMP-1, and TIMP-1 expressions results in sclerotic skin disease, such as scleroderma, hypertrophic scar, and keloids. Curcumin, a phytochemical extracted from the rhizomes of Curcuma longa, showed an anti-fibrotic effect in an animal study, such as a pulmonary and cholangioductal fibrosis animal model. However, the expressions of type I collagen, MMP-1, Smad2/3, and TIMP-1 in curcumin treated human skin fibroblasts is largely unknown. OBJECTIVE: The purpose of this study was to investigate the expressions of type I collagen, MMP-1, Smad2/3, and TIMP-1 proteins in curcumin-treated human skin fibroblasts. METHODS: Human skin fibroblasts were treated by various concentrations of curcumin (1~40 uM). The expressions of type I collagen, MMP-1, Smad2/3, and TIMP-1 proteins were analyzed by Western blot analysis. In addition, activities of type I collagen promoter were analyzed by the CAT assay. RESULTS: The expression of type I collagen decreased but the expression of MMP-1 and TIMP-1 increased by curcumin treatment in a dose and time dependent manner in Western blot analysis. Type I collagen promoter activities were decreased by curcumin treatment in the CAT assay. Smad2/3 expression decreased by curcumin treatment but TGF-beta1 induced Smad2/3 activation was not decreased by curcumin treatment following Western blot analysis. CONCLUSION: Decrease of type I collagen expression through the inhibition of Smad2/3 and increase of the expression of MMP-1, which is the degradating enzyme of type I collagen, in human skin fibroblasts by curcumin treatment offer expectation of curcumin as antifibrotic agent.

The Efficacy of Shikonin on Cartilage Protection in a Mouse Model of Rheumatoid Arthritis

The potential therapeutic action of shikonin in an experimental model of rheumatoid arthritis (RA) was investigated. As a RA animal model, DBA/1J mice were immunized two times with type II collagen. After the second collagen immunization, mice were orally administered shikonin (2 mg/kg) once a day for 35 days, and the incidence, clinical score, bone mineral density (BMD), bone mineral content (BMC) and joint histopathology were evaluated. BMD in the proximal regions of the tibia largely increased in the shikonin treatment group compared with the control group. We also examined the effect of shikonin on inflammatory cytokines and cartilage protection. Shikonin treatment significantly reduced the incidence and severity of collagen-induced arthritis (CIA), markedly abrogating joint swelling and cartilage destruction. Shikonin also significantly inhibited the production of matrix metalloproteinase (MMP)-1 and up-regulated tissue inhibitors of metalloproteinase (TIMP)-1 in mice with CIA. In conclusion, shikonin exerted therapeutic effects through regulation of MMP/TIMP; these results suggest that shikonin is an outstanding candidate as a cartilage protective medicine for RA.

Cholesterol, a Major Component of Caveolae, Down-regulates Matrix Metalloproteinase-1 Expression through ERK/JNK Pathway in Cultured Human Dermal Fibroblasts

BACKGROUND: Cholesterol is a major component of specialized membrane microdomains known as lipid rafts or caveolae, which modulate the fluidity of biological membranes. Membrane cholesterol therefore plays an important role in cell signaling and vesicular transport. OBJECTIVE: In this study, we investigated the effects of cholesterol on matrix metalloproteinase-1 (MMP-1) expression in human dermal fibroblasts. METHODS: MMP-1 mRNA and protein expression were determined by RT-PCR and Western blotting, respectively. AP-1 DNA binding activity was detected by electrophoretic mobility shift assays. The amount of cholesterol was analyzed by cholesterol assay kit. RESULTS: We observed that MMP-1 mRNA and protein expression was dose-dependently decreased by cholesterol treatment. In contrast, cholesterol depletion by a cholesterol depletion agent, methyl-beta-cyclodextrin (M beta CD) in human dermal fibroblasts, increased MMP-1 mRNA and protein expression in a dose-dependent manner. Also, we investigated the regulatory mechanism of M beta CD-induced MMP-1 expression: cholesterol depletion by M beta CD, activated ERK1/2 and JNK, but not p38 MAPK cascade, and it also significantly increased c-Jun phosphorylation, c-Fos expression and activator protein-1 binding activity. Furthermore, the inhibition of ERK or JNK with specific chemical inhibitors prevented M beta CD-induced MMP-1 expression, which indicates that ERK and JNK play an important role in cholesterol depletion-mediated MMP-1 induction. In addition, M beta CD-induced phosphorylation of ERK and JNK and MMP-1 expression were suppressed by cholesterol repletion. CONCLUSION: Our results suggest that cholesterol regulates MMP-1 expression through the control of ERK and JNK activity in human dermal fibroblasts.