ABSTRACT
Objective: To investigate the main mechanisms of pulmonary fibrosis following silica nanoparticles (SiNPs) exposure through constructing the macrophage-fibroblast model in vitro, which simulated the process of pulmonary fibrosis. Methods: In January 2021, human mononuclear leukemia cells (THP-1) were treated with 0, 25, 50, 100 μg/ml SiNPs for 24 h. The supernatant of THP-1 cells was collected and applied to human embryonic lung fibroblast cells (MRC-5) which divided into control and low, medium and high dose groups at the logarithmic growth stage for 24 h. MRC-5 cell viability was detected by CCK8. The hydroxyproline (Hyp), interleukin 6 (IL-6), interleukin 1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) expression were detected in the supernatants of MRC-5. The changed proteins were detected by liquid-phase mass spectrometry in high dose group. GeneCard database were applied to identity the differential pulmonary fibrosis proteins in high dose group. Gene Ontology (GO) was performed to identity the key biological process in differential pulmonary fibrosis proteins of high dose group. The String database was used to construct the protein-protein interactions (PPI) network of differential pulmonary fibrosis proteins. The APP of CytoHubba was applied to calculate the key protein of differential pulmonary fibrosis proteins in PPI network. Correlation coefficients between key differential pulmonary fibrosis proteins were calculated using Pearson correlation analysis. Western blotting was applied to detect the expression of key proteins of differential pulmonary fibrosis proteins in different groups. Results: CCK8 results showed that MRC-5 cell viability was increasing in low, medium and high dose groups compared with control group (P<0.05). The expression levels of Hyp and IL-1β in different group were increased compared with control group, the expression levels of IL-6 and TNF-α were increased in high dose group compared with control group (P<0.05). GeneCard database identified 26 differential pulmonary fibrosis proteins, which were mainly involved in extracellular matrix hydrolysis, cell inflammatory response, tissue repair, cell proliferation, inflammation response by GO analysis. The APP of CytoHubba was calculated that matrix metalloproteinase 9 (MMP9) and tissue inhibitor metalloproteinase 1 (TIMP1) played an important role in PPI network. The results of correlation analysis showed that MMP9 was correlated with the expression of matrix metalloproteinase 1 (MMP1), matrix metalloproteinase 3 (MMP3), TIMP1 and epidermal growth factor receptor (EGFR) (r=0.97, 0.98, 0.94, 0.93, P<0.05). Western blotting results showed that TIMP1 protein expression was increased in low, medium and high dose groups, while MMP9 protein expression was increased only in high dose group (P<0.05) . Conclusion: Differential expression proteins related with pulmonary fibrosis in MRC-5 cells mainly regulate biological processes of extracellular matrix hydrolysis, tissue repair, and cellular inflammation response following SiNPs exposure. MMP9 and TIMP1 may be the key proteins, which affected the fibrosis process in vitro pulmonary fibrosis model.
ABSTRACT
Background: It is well established that chronic exposure to tobacco induces head and neck cancers but the exact etiopathogenesis is not known. Though studies have shown expression of TIMP1, EPS8 and AXL in cancers, their role in tobacco-induced cancers is not known. We aimed this study to evaluate the role of these molecules in oral and oropharyngeal squamous cell cancers (SCC). Materials and Methods: In this single institutional study, 31 patients of oral and oropharyngeal SCC with history of chewing tobacco were included. Smokers were excluded from the study. After informed consent biopsies were taken from affected and contralateral normal mucosa. Paraffin blocks were made and tissue microarray (TMA) were constructed using these blocks. Immunohistochemistry (IHC) for TIMP1, EPS8, AXL kinase was carried out on these tissue microarrays. The intensity of staining was scored from 0 to 3+, related to expression of each of the three molecules. Results: The expression of TIMP1, EPS8 and AXL kinase was significantly more in the cancerous mucosa versus non-cancerous mucosa (P = 0.000 in all three) in oral and oropharyngeal SCC exposed to chewing tobacco. Conclusion: Immunohistochemical expression of these molecular markers in oral and oropharyngeal SCC correlated with their molecular based studies. Significant IHC expression of TIMP1, EPS8 and AXL establishes their role in the pathogenesis of oral and oropharyngeal squamous cell carcinomas. Novel targeted therapies may be researched that can detect and target these molecules at an earlier stage of pathogenesis of these tumors.
ABSTRACT
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.
ABSTRACT
Objective: To observe the effects of different combinations of Gentianae Macrophyllae Radix (Qinjiao) on the ankle joint matrix metalloproteinase-3 (MMP-3) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) of rheumatoid arthritis (RA) model rats with wind-cold-dampness arthralgia.Method: Eighty healthy SD rats were randomly divided into 8 groups, namely blank control group, collage Ⅱ model group, wind-cold-dampness syndrome model group, positive control group, single-taste Gentianae Macrophyllae Radix group, Gentianae Macrophyllae Radix-Clematidis Radix et Rhizoma group (GC group), Gentianae Macrophyllae Radix-Taxilli Herba group (GT group), Gentianae Macrophyllae Radix-Stephanlae Tetrandrae Radix group (GS group), with 10 rats in each group. Rat model of wind-cold-dampness RA was induced through the injection with type Ⅱ collagen emulsion and wind-cold-dampness stimulation. After the establishment of the model, the blank control group, collage Ⅱ model group and wind-cold-dampness syndrome model group were given normal saline, and the corresponding liquid medicine was given to each administration group. In the experiment, the thickness of the left posterior metatarsal of rats was measured every 3 days, and the swelling degree of metatarsal was calculated. The arthritis index (AI) was evaluated on the 38th day of the experiment. The serum rheamatoid factor(RF) content of rats was detected by enzyme linked immunosorbent assay (ELISA). The expressions of MMP-3 and TIMP-1 in ankle joint were detected by Western blot. The expressions of MMP-3 and TIMP-1 mRNA in ankle joint were detected by real-time fluorescence quantitative PCR (Real-time PCR).Result: Compared with the blank group, the swelling degree, AI score, serum RF content, MMP-3 protein expression and MMP-3 mRNA expression in ankle joints of coll age Ⅱ model group and model wind-cold-dampness syndrome group were significantly increased (PPPPPPConclusion: For rheumatoid arthritis with wind-cold-dampness arthralgia, mild and warm traditional Chinese medicine (TCM) has a better effect than the combination of mild and cold TCM or mild TCM drugs. The experimental results are basically consistent with the principle of "treating cold diseases with hot medicine". The mechanism of the compatibility in treating rheumatoid arthritis due to wind-cold-dampness arthralgia may be related to the reduction of MMP-3, the increase of TIMP-1 expression and the reduction of articular cartilage damage.
ABSTRACT
OBJECTIVE： To study the effects of Dengzhan shengmai capsules combined with isosorbide mononitrate on serum TIMP-1 and MMP-9 levels， blood lipid level and cardiac function in patients with unstable angina pectoris （UAP）. METHODS： Totally 198 UAP patients admitted to our hospital from Apr. 2016 to Apr. 2019 were selected， and divided into observation group （n＝102） and control group （n＝96） according to therapy plan. Control group received Isosorbide mononitrate tablets 40 mg， qd， orally； observation group additionally received Dengzhan shengmai capsules， 2 capsules per time， tid， orally， on the basis of control group. Both groups received treatment for consecutive 4 weeks. The clinical efficacy， serum levels of MMP-9 and TIMP-1， blood lipid indexes [low density lipoprotein cholesterol （LDL-C）， high density lipoprotein cholesterol （HDL-C）， triglyceride （TG）， total cholesterol （TC）]， cardiac function indexes [end-diastolic volume （EDV）， left ventricular ejection fraction （LVEF）， end-systolic volume （ESV）] and ADR were compared between 2 groups. RESULTS： The total response rate of observation group was significantly higher than control group （91.18％ vs. 70.83％， P＜0.05）. Before treatment， there was no statistical significance in the levels of ESV， MMP-9， TIMP-1， TC， LDL-C， TG， HDL-C， EDV and LVEF between 2 groups （P＞0.05）. After treatment， the levels of MMP-9， TC， LDL-C， TG， ESV and EDV in 2 groups were decreased significantly in both groups， and the observation group was significantly lower than the control group （P＜0.05）； meanwhile， the levels of TIMP-1， HDL-C and LVEF in 2 groups were increased significantly， and the observation group was significantly higher than control group （P＜0.05）. There was no statistical significance in the incidence of total ADR in 2 groups （P＞0.05）. CONCLUSIONS： Dengzhan shengmai capsules combined with isosorbide mononitrate show good clinical efficacy for UAP， which can effectively reduce serum MMP-9 level and increase serum TIMP-1 level， reduce blood lipid levels， as well as improve cardiac function for UAP patients， with good safety.
ABSTRACT
Objective To investigate the significance of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in serum and cerebrospinal fluid for evaluation of severe hand ,foot ,and mouth disease (HFMD) complicated with neurogenic pulmonary edema (NPE).Methods A total of 140 patients diagnosed with HFMD in Henan Children′s Hospital were enrolled and divided into three groups including mild group ,severe HFMD group without NPE ,severe HFMD group with NPE .These severe HFMD patients were also divided into survival group and death group according to the 28-day prognosis .Meanwhile ,50 age-matched healthy children were selected as controls .Serum MMP-9 and TIMP-1 levels were measured in all enrolled children .At the same time ,MMP-9 ,TIMP-1 and ratio of MMP-9／TIMP-1 in cerebrospinal fluid were measured in the severe HFMD group with and without NPE .Quantitative data were compared using one-way analysis of variance , and means comparisons between samples were conducted using LSD-t test .Results Among 140 children with HFMD ,66 were in mild group ,42 in severe HFMD without NPE group ,and 32 in severe HFMD with NPE group .And 50 healthy children were in control group .After 28 days ,14 cases died in severe HFMD groups .MMP-9 , TIMP-1 and MMP-9／TIMP-1 in serum of severe HFMD group with NPE increased significantly greater than those in the other three groups (F＝269 .356 ,121 .301 and 101 .502 ,respectively ,all P ＜0 .05). MMP-9 ,TIMP-1 and MMP-9／TIMP-1 in cerebrospinal fluid of severe HFMD group with NPE were (57 .24 ± 8 .92) μg／L ,(35 .26 ± 8 .14) μg／L and (1 .66 ± 0 .23) μg／L ,respectively ,while those in cerebrospinal fluid of severe HFMD group without NPE were (30 .57 ± 3 .89) μg／L ,(26 .25 ± 0 .32) μg／L and (1 .17 ± 0 .61) μg／L ,respectively .The differences between the two groups were all statistically significant (t＝62 .485 ,37 .680 and 169 .387 ,respectively ,all P＜0 .01).MMP-9 ,TIMP-1 and MMP-9／TIMP-1 in serum and cerebrospinal fluid of death group increased significantly greater than those in survival group ,the difference were statistically significant (all P＜0 .01).The maximum area under curve (AUC) was reached when the MMP9／TIMP-1 ratio in cerebrospinal fluid was 0 .890 (95% CI :0 .801 －0 .978).Conclusions MMP-9 and TIMP-1 may be involved in the pathogenesis of HFMD complicated with NPE .The detection of MMP-9 and TIMP-1 levels may be beneficial for the early diagnosis of severe HFMD with NPE .The imbalance of MMP-9／TIMP-1 ratio can be used as one of the predictors of severe HFMD combined with NPE.
ABSTRACT
Objective: To explore the possible effects and mechanism of modified Compound Biejia Ruangan Pills (MF-CBRP) on liver fibrosis induced by CCl4 in rats. Methods: SD rats were randomly divided into control group, model group, positive control (colchicine, 0.1 mg/kg) group, CBRP group (0.594 7 g/kg), MF-CBRP high-and low-dose (1.061 0 and 0.530 5 g/kg) groups. The rat model with liver fibrosis was established by sc injection CCl4 solution (dissolved in soybean oil), twice a week for six weeks except control group. The rats in the treatment groups were administered six weeks after the model establishment. At the end of the administration, the contents of serum biochemical, hydroxyproline (Hyp), lipid peroxidation, fibrosis factor, platelet-derived factor-α (PDGF-α), and connective tissue growth factor (CTGF) were measured and the pathological changes of liver tissue were examined by HE and Masson. The level of α-SMA and the expression of transforming growth factor-β1 (TGF-β1), matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-1 (TIMP-1) were detected by immunohistochemistry and RT-PCR. Results: The results showed that MF-CBRP improved the liver function significantly through reducing the level of ALT, AST, T-Bil, ALP, Hyp, HA, LN, PC-III, IV-C, and MDA (P < 0.05, 0.01), and increasing the content of S/L, TP, ALB, SOD, and GSH (P < 0.05, 0.01). MF-CBRP also reduced the expression of PDGF-α, CTGF, α-SMA, MMP-2, and TIMP-1 mRNA (P < 0.01) and improved the pathological changes of liver histopathology. Compared with CBRP, MF-CBRP anti-hepatic fibrosis effect was slightly weakened. Conclusion: The results suggested that MF-CBRP may against hepatic fibrosis by reducing oxidative stress, inhibiting collagen fibrillation, reversing hepatic stellate cell activation, and inhibiting the expression of hepatic growth factor. However its anti-fibrosis activity and mechanism was weaker than the original CBRP in a certain extent.
ABSTRACT
Aim To investigate the effects of IcarisideⅡ ( ICS Ⅱ) on myocardial fibrosis in spontaneously hypertensive rat( SHR) . Methods Twenty male SHR rats were randomly divided into the model group (group SHR) , ICS Ⅱ low ( ICS Ⅱ-L) , middle ( ICSⅡ-M) and high ( ICS Ⅱ-H) group, and male WKY rats were set as control group ( group WKY) . ICS Ⅱ-L, ICSⅡ-M and ICSⅡ-H groups were intragastrically administered with ICS Ⅱ for 12 weeks. After that the blood pressure was measured in rats. Then, the rats were sacrificed and the left ventricles were separated in order to calculate the left ventricular mass index. Mas-son staining was used to detect the occurrence of inter-stitial fibrosis in cardiac tissues. Real time PCR was used to observe the gene expression of MMP-2, MMP-9 and TIMP-1 in the left ventricle in SHRs. The protein expression levels of MMP-2, MMP-9, TIMP-1, Colla- gen Ⅰ and Collagen Ⅲ were measured by Western blot. Results Compared with SHR group, the myo-cardial fibrosis was reduced after ICS Ⅱ (8, 16 mg· kg-1) treatment. The blood pressure and left ventricu-lar mass index decreased(P<0.05). The expressions of MMP-2, MMP-9, CollagenⅠand CollagenⅢwere down-regulated in left ventricular tissues( P <0.05 ) , while the expression of TIMP-1 was up-regulated( P<0.05) . Conclusion Icariside Ⅱ ameliorates myocar-dial fibrosis in SHR, and the mechanisms might be re-lated to the decrease of blood pressure and down-regu-lation of MMP-2, MMP-9 expression and up-regulation of TIMP-1 expression.
ABSTRACT
OBJECTIVE:To investigate the effects of polydatin on protein expression of MMP-9 and TIMP-1 in renal tissue of renal fibrosis model rats,and to investigate the mechanism of delaying renal fibrosis in rats. METHODS:A total of 40 rats were randomly divided into sham operation group(normal saline),model group(normal saline),polydatin group(100 mg/kg)and benazepril group(positive control,5 mg/kg). Except for sham operation group,renal fibrosis rat model was induced by unilateral ureteral obstruction in other groups. 1 h after surgery,each group was given relevant medicine intragastrically,once a day.After 4 weeks of consecutive treatment,the contents of β 2-microglobulin(β 2-MG)and N-acetyl-β-D-glucosaminidase(NAG)in urine, serum contents of BUN and Cr were detected in each group. HE staining was used to observe and score pathological changes of renal tissue in rats. Protein expressions of MMP-9 and TIMP-1 in renal tissue of rats were determined by immunohistochemistry. RESULTS:Compared with sham operation group,the contents of β2-MG and NAG in urine,serum contents of BUN and Cr were increased significantly in model group(P<0.05),and pathological score and protein expressions of MMP-9 and TIMP-1 were also increased significantly(P<0.01). Compared with model group,protein expression of MMP-9 in renal tissue were increased continuously in polydatin group and benazepril group(P<0.01);other indexes were decreased significantly(P<0.05 or P<0.01),and there was no statistical significance in each index between polydatin group and benazepril group(P>0.05). CONCLUSIONS:Polydatin can delay renal fibrosis,the mechanism of which may be associated with up-regulating protein expression of MMP-9,down-regulating protein expression of TIMP-1 and increasing the ratio of MMP-9/TIMP-1.
ABSTRACT
Objective: To observe the effects of He-Ne laser on the expression of extracellular matrix and its regulatory factors in degenerated temporomandibular joint(TMJ) of rabbits. Methods: 40 New Zealand adult white rabbits were randomly divided into normal group,sham model group,TMJOA model group and laser treatment group(n = 10),the rats in treatment group were treated by He-Ne laser irradination at acupoints. The rats in each group were divided into 1 d and 11 d groups(n = 5). The animals were respectively killed 1 d and 11 d after operation and HE staining was used to observe the histomorpholy. The protein expression of extracellular matrix and its regulatory factors were examined by Western blot. Results: After He-Ne laser treatment,the fiber layer of condylar cartilage was slightly loose,part of the fiber was newly produced. The level of Col-2,PRG-4,TIMP-1,BMP-2 was up-regulated and the of MMP-13 was down-regulated in the 11 d treatment group. Conclusion: He-Ne laser irradiation on acupoints may up-regulate the expression of extracellular matrix (Col-2 and PRG-4) and its regulatory factors (TIMP-1 and BMP-2),down-regulate the expression of MMP-13.
ABSTRACT
ABSTRACT Background: Gastric cancer is the 3rd most common cause of death in men and the 5th common in women worldwide. Today, surgery is the only curative therapy. Currently available advanced imaging modalities can predict R0 resection in most patients, but it can only be detected with certainty in the perioperative period. Aim: To determine the role of serum CK18, MMP9, TIMP1 levels in predicting R0 resection in patients with gastric cancer. Methods: Fifty consecutive patients scheduled for curative surgery with gastric adenocarcinoma diagnosed between 2013-2015 were included. One ml of blood was taken from the patients to analyze CK18, MMP9 and TIMP1. Results: CK18, MMP9 and TIMP1 levels were positively correlated with pathological N and the stage (p<0,05). CK-18, MMP-9 and TIMP-1 averages in positive clinical lymph nodes and in clinical stage 3, were found to be higher than the averages of those with negative clinical lymph nodes and in clinical stage 2 (p<0,05). Conclusion: Although serum CK-18, MMP-9 and TIMP-1 preoperatively measured in patients scheduled for curative surgery did not help to evaluate gastric tumor resectability, they were usefull in predicting N3-stage.
RESUMO Racional: Câncer gástrico é a terceira causa mais comum de morte em homens e a quinta em mulheres em todo o mundo. Atualmente a cirurgia é a única terapia curativa. As modalidades de imagem avançadas atualmente disponíveis podem prever a ressecção R0 na maioria dos pacientes, mas ela só pode ser detectada durante o perioperatório. Objetivo: Determinar o papel dos níveis séricos de CK18, MMP9 e TIMP1 na predição da ressecção R0 em pacientes com câncer gástrico. Métodos: Foram incluídos no estudo pacientes consecutivos agendados para operação curativa entre 2013-2015. Foi retirado 1 ml de sangue dos pacientes incluídos para estudar CK18, MMP9 e TIMP1. Resultados: Os níveis de CK18, MMP9 e TIMP1 foram positivamente correlacionados com o N patológico e o estadiamento (p<0,05). As médias CK-18, MMP-9 e TIMP-1 das pessoas com linfonodos positivos e aqueles em estágio clínico 3 foram superiores às médias das pessoas com linfonodos negativos e estágio clínico 2 (p<0,05). Conclusão: Embora as dosagens séricas de CK-18, MMP-9 e TIMP-1 em pacientes agendados para operação curativa por adenocarcinoma gástrico não ajudem a ter ideia de ressecabilidade tumoral, ela foi útil na predição de estadiamento N3.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Aged, 80 and over , Stomach Neoplasms/surgery , Stomach Neoplasms/blood , Adenocarcinoma/surgery , Adenocarcinoma/blood , Matrix Metalloproteinase 9/blood , Keratin-18/blood , Reference Values , Stomach Neoplasms/pathology , Adenocarcinoma/pathology , Biomarkers, Tumor/blood , Logistic Models , Statistics, Nonparametric , Tissue Inhibitor of Metalloproteinase-1/blood , Lymphatic Metastasis/pathology , Neoplasm StagingABSTRACT
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.
ABSTRACT
AIM To study the effects of Biejiajian Pills (Colla Carapacis Trionycis,Asini Corii Colla,Nidus Vespae,etc.) on NF-κB,p65,p50 and IκB in NF-κB signaling pathway and target gene expression in HSC-T6 cells of rats.METHODS HSC-T6 cells were cultured with Biejiajian Pills drug serum for 24 hours,the expressions of p65,p50,VEGF and TIMP-1 mRNA were determined by qPCR;the expression of p65 was measured by immunofluorescence;the expressions of IκBα,IκBβ and α-SMA were determined by Western blot.RESULTS The Biejiajian Pills middle-,high-dose and positive control groups showed significantly lower expressions of p65,VEGF and TIMP-1 mRNA as compared with the blank control group and negative control group,the expressions of p50 mRNA among various groups showed no significant differences.But immunofluorescence showed that the expression of p65 in cytoplasm was decreased.Meanwhile,Biejiajian Pills showed significantly higher IκBα protein expression and obvious down-regulation of α-SMA expression in a dose-dependent manner,but had no significant influence on the expression of IκBβ.CONCLUSION Biejiajian Pills' therapeutic effects on hepatic fibrosis may be related to influencing NF-κB signaling pathway and inhibiting the expression of down-stream target gene.
ABSTRACT
Objective:To investigate the effects and mechanisms of Xuebijing injection on early sepsis induced acute lung injury and explore a new therapy.Methods:Thirty healthy male Sprague-Dawley rats were randomly divided into 3 groups:Sham group(n=10),NS group (NS 4mL/kg n=10),Xuebijing Group(Xuebijing 4mL/kg n=10).Cecal ligation and puncture (CLP) was used to duplicate severe sepsis model.At 6h after CLP,the rats were sacrificed,the lungs were resected and histopathological characteristic was observed by transmission electron microscopy technique.The change of the lung wet/dry (W/D) weight ratio were studied.Meanwhile,the lungs were resected for detection of ET-1,iNOS,MMP-9,TIMP-1 mRNA with reverse transcription-polymerase chain raction (RT-PCR).Results:The changes of pulmonary alveoli and the interstitial edema as well as lung tissue in Xuebijing group were better than those of NS group.The change of lung wet/dry (W/D) weight ratio in septic rats showed was significantly increased at 6 h after CLP (vs Sham group:5.37± 0.12 vs 4.33± 0.06,P＜0.01).The lung W/D was significantly decreased in XBJ group (vs NS group:4.67± 0.09 vs 5.37± 0.12,P＜0.05).The expressions in XBJ group lung tissue of ET-1 (0.511 ± 0.111 vs 0.705± 0.122,P＜0.01),iNOS(0.45610.075 vs 0.548± 0.098,P＜0.05)、MMP-9 (0.617± 0.079 vs 0.732± 0.131,P＜0.05),TIMP-1 (0.438± 0.043 vs 0.515± 0.049,P ＜0.01) mRNA were significantly decreased than those in NS group.And the expressions of ET-1 (0.705± 0.122vs 0.400± 0.033,P＜0.01),iNOS (0.548± 0.098 vs 0.334± 0.027,P＜0.01),MMP-9(0.732± 0.131 vs 0.352± 0.061,P＜0.01),TIMP-1(0.515± 0.049 vs 0.365± 0.068,P＜0.01) mRNA in NS group were significantly increased compared with those in the sham group.Conclusions:Xuebijing could protect against sepsis induced acute lung injury,which might be related with the decrease ofET-1,iNOS,MMP-9 and TIMP-1 mRNA expressions.
ABSTRACT
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.
ABSTRACT
Objective: To observe the effects of Gentianae Macrophyllae Radix (Qinjiao) in different combinations on the expression of MMP-3, TIMP-1, and ankle pathological changes in rheumatoid arthritis (RA) model rats, and to explore the relationship between medicinal properties-disease-efficacy and explore the mechanism of traditional Chinese medicine in the treatment of RA. Methods: Eighty SD rats were randomly divided into blank group, collage II model group, rheumatic fever model group, tripterygium group, single Qinjiao group, Qinjiao-Weilingxian (Clematidis Radix et Rhizoma) group, Qinjiao-Sangjisheng (Taxilli Herba) group, and Qinjiao-Fangji (Stephanlae Tetrandrae Radix) group. Rheumatic fever arthralgia model was induced by collage II and exposed in rheumatic fever environment. After modeling, each administration group was ig administrated with corresponding drug solution. The paw thickness was observed every 3 d, and swelling was calculated; The arthritis index was scored in the early, middle, and late stages. Rats were sacrificed on day 39. The expression of MMP-3 and TIMP-1 in the ankle joint of rats was detected by immunohistochemistry, and the pathological changes of ankle joint were observed by HE staining. Results: Compared with the blank group, the paw edema, AI, and expression of MMP-3 in collage II model group and rheumatic fever model group was significantly higher, the expression of TIMP-1 were significantly lower, the articular surface of two groups was rough and damaged, and the articular cartilage was severely damaged. A large number of granulation tissues proliferated, accompanied by a large number of inflammatory cell infiltration and neovascularization. After treatment, compared with rheumatic fever model group, the paw edema, AI, and expression of MMP-3 in all treatment groups were reduced to varying degrees, in which the Qin-Fang group were lower most significant; The serum levels of TNF-α and IL-1β in all treatment groups were significantly decreased in varying degrees, especially in the Qin-Fang group; The expression of TIMP-1 was significantly increased in varying degrees, especially in the Qin-Fang group. HE staining results showed that the area of articular cartilage destruction and the degree of destruction were reduced, the inflammatory cells and neovascularization decreased, the number of repair fibers increased, and the scar tissue increased. Conclusion: For the rheumatic fever arthralgia RA, the combined effect of mild and cold Chinese materia medica (CMM) is better than the combined effect of mild and warm CMM, the combined effect of mild CMMs. Experimental results are consistent with the principles of traditional Chinese medicine clinical treatment of "treating the hot diseases should use the cold medicine", the mechanism of the compatibility drug relieving rheumatic fever arthralgia RA may be related to its ability, which can reduce the expression of MMP-3, increase the expression of TIMP-1, reduce articular cartilage destruction, and reduce inflammatory cell infiltration and vascular proliferation.
ABSTRACT
Objective To investigate the effects of all-trans retinioc acid (ATRA)on proliferation of rat hepatic stellate cells (HSC-T6)and expressions of collagen Ⅰ,matrix metalloproteinase-2 (MMP-2),tissue inhibitor of metalloproteinases-1 (TIMP-1 )and signal protein Smad2/3 in TGF-β1-simulated HSC-T6 so as to explore the impact and molecular mechanisms of ATRA on liver fibrosis in vitro .Methods Cultured HSC-T6s were treated with different concentrations of ATRA (0.1,1,10 μmol/L)for fixed time (12,24,48 hours).After intervention time,cell proliferation was evaluated by MTT.Meanwhile,HSC-T6s stimulated by TGF-β1 (5 ng/mL)were treated with different concentrations of ATRA for 24 h.The mRNA expressions of COL1α2,MMP-2 and TIMP-1 were quantified by RT-PCR;the expression of Smad 2/3 protein was determined by cell immunochemistry.Results The proliferation of hepatic stellate cells was inhibited by ATRA in a dose-dependent manner (P < 0.05 ).After induced by TGF-β1,the mRNA expressions of COL1α2,MMP-2 and TIMP-1 and the expression of Smad 2/3 protein were increased significantly compared with control group (P <0.05).However,ATRA could obviously reduce themRNA expressions of COL1α2,MMP-2 and TIMP-1 and the expression of Smad 2/3 protein in HSC-T6 induced by TGF-β1 (P < 0.05 ).Conclusion ATRA can inhibit the proliferation of HSC-T6s and reduce the mRNA expressions of COL1α2,MMP-2 and TIMP-1 in HSC-T6 which were induced by TGF-β1.The anti-hepatic fibrosis function of ATRA may be related to its inhibition on the expression of Smad 2/3 protein in HSC-T6 to influence TGF-β1/Smad signaling pathway.
ABSTRACT
Objective To investigate the effects of all-trans retinioc acid (ATRA)on proliferation of rat hepatic stellate cells (HSC-T6)and expressions of collagen Ⅰ,matrix metalloproteinase-2 (MMP-2),tissue inhibitor of metalloproteinases-1 (TIMP-1 )and signal protein Smad2/3 in TGF-β1-simulated HSC-T6 so as to explore the impact and molecular mechanisms of ATRA on liver fibrosis in vitro .Methods Cultured HSC-T6s were treated with different concentrations of ATRA (0.1,1,10 μmol/L)for fixed time (12,24,48 hours).After intervention time,cell proliferation was evaluated by MTT.Meanwhile,HSC-T6s stimulated by TGF-β1 (5 ng/mL)were treated with different concentrations of ATRA for 24 h.The mRNA expressions of COL1α2,MMP-2 and TIMP-1 were quantified by RT-PCR;the expression of Smad 2/3 protein was determined by cell immunochemistry.Results The proliferation of hepatic stellate cells was inhibited by ATRA in a dose-dependent manner (P < 0.05 ).After induced by TGF-β1,the mRNA expressions of COL1α2,MMP-2 and TIMP-1 and the expression of Smad 2/3 protein were increased significantly compared with control group (P <0.05).However,ATRA could obviously reduce themRNA expressions of COL1α2,MMP-2 and TIMP-1 and the expression of Smad 2/3 protein in HSC-T6 induced by TGF-β1 (P < 0.05 ).Conclusion ATRA can inhibit the proliferation of HSC-T6s and reduce the mRNA expressions of COL1α2,MMP-2 and TIMP-1 in HSC-T6 which were induced by TGF-β1.The anti-hepatic fibrosis function of ATRA may be related to its inhibition on the expression of Smad 2/3 protein in HSC-T6 to influence TGF-β1/Smad signaling pathway.
ABSTRACT
This study was to evaluate, the pro-apoptotic activity of ethanolic Chelidonium majus L. (CM) leaf extract and/or gamma-radiation in Ehrlich ascites carcinoma (EAC) bearing mice by measuring tumor volume, apoptotic factors (caspase-3, Bcl-2 and Bax proteins, tumor necrosis factor-alpha (TNF-), angiogenesis factors (matrix metalloproteinase (MMP-2 and MMP-9); tissue inhibitor of metalloproteinases (TIMP-1), DNA fragmentation, besides histopathological examination. After tumor inoculation, mice received CM (100 mg/body weight/day) on the 10th day and were exposed to whole body -radiation (6Gy) on the 17th day. The data obtained reveales that combining CM with - radiation exposure induce significant regression in tumor growth, up-regulate Bax, caspase-3, TIMP-1 and inhibit Bcl-2, TNF-, MMP-(2 and 9). Cytotoxicity is substantiated by increased DNA-fragmentation (comet assay and DNA content) and histological examination. It could be suggested that combining CM with gamma-radiation increased apoptosis of tumor cells which would help to increase the lifespan of mice.
Este estudo foi realizado para avaliar a atividade pró-apoptótica do extrato etanólico da folha da Chelidonium majus L. (CM) e/ou radiação gama no tumor ascítico de Ehrlich tendo ratos para a medição do volume do tumor, fatores apoptóticos (caspase-3, proteínas Bcl-2 e Bax, fator de necrose tumoral (TNF-)), fatores angiogênicos (metaloproteinase matriz (MMP-2 e MMP-9)); inibidor tecidual de metaloproteinase (TIMP-1), fragmentação de DNA, além de exame histopatológico. Depois da inoculação do tumor, os ratos receberam CM 100 mg/peso corporal/dia no 10º dia e tiverem o corpo completo exposto a radiação gama (6Gy) no 17º dia. Os dados obtidos revelam que combinar CM com exposição à radiação gama induz a uma regressão significativa no crescimento do tumor, regula positivamente Bax, caspase-3, TIMP-1 e inibe Bcl-2, TNF-, MMP-(2 e 9). Citotoxicidade é substanciada pelo aumento da fragmentação do DNA (ensaio cometa e conteúdo de DNA) e exame histopatológico. Poderia ser sugerido que combinar CM com radiação gama aumentou a apoptose das células tumorais o que ajudaria a aumentar a expectativa de vida dos ratos.
Subject(s)
Rats , Carcinoma, Ehrlich Tumor , Chelidonium , Caspase 3 , DNA FragmentationABSTRACT
Background & objectives: Ischaemic stroke is a life burdening disease for which carotid endarterectomy (CEA) is considered a gold standard intervention. Pro-inflammatory markers like matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) and S-100 Beta (S100B) may have a role in the early inflammation and cognitive decline following CEA. This study was aimed to describe the perioperative time courses and correlations between of MMP-9, TIMP-1 and S100B following CEA. Methods: Fifty four patients scheduled for CEA were enrolled. Blood samples were collected at four time points, T1: preoperative, T2: 60 min after cross-clamp release, T3: first postoperative morning, T4: third postoperative morning. Twenty atherosclerotic patients were included as controls. Plasma MMP-9, TIMP-1 and S100B levels were estimated by ELISA. Results: TIMP-1 was decreased significantly in the CEA group (P<0.01). Plasma MMP-9 was elevated and remained elevated from T1-4 in the CEA group (P<0.05) with a marked elevation in T3 compared to T1 (P<0.05). MMP-9/TIMP-1 was elevated in the CEA group and increased further by T2 and T3 (P<0.05). S100B was elevated on T2 and decreased on T3-4 compared to T1. Interpretation & conclusions: Our study provides information on the dynamic changes of MMP-9-TIMP-1 system and S100B in the perioperative period. Preoperative reduction of TIMP-1 might be predictive for shunt requirement but future studies are required for verification.