Jiming Powder is a traditional ancient prescription with good therapeutic effect in the treatment of heart failure, but its mechanism lacks further exploration. In this study, a mouse model of coronary artery ligation was used to evaluate the effect and mechanism of Jiming Powder on myocardial fibrosis in mice with myocardial infarction. The study constructed a mouse model of heart failure after myocardial infarction using the method of left anterior descending coronary artery ligation. The efficacy of Jiming Powder was evaluated from multiple angles, including ultrasound imaging, hematoxylin-eosin(HE) staining, Masson staining, Sirius Red staining, and serum myocardial enzyme spectrum detection. Western blot analysis was performed to detect key proteins involved in ventricular remodeling, including transforming growth factor-β1(TGF-β1), α-smooth muscle actin(α-SMA), wingless-type MMTV integration site family member 3a(Wnt3a), β-catenin, matrix metallopeptidase 2(MMP2), matrix metallopeptidase 3(MMP3), TIMP metallopeptidase inhibitor 1(TIMP1), and TIMP metallopeptidase inhibitor 2(TIMP2). The results showed that compared with the model group, the high and low-dose Jiming Powder significantly reduced the left ventricular internal diameter in systole(LVID;s) and diastole(LVID;d), increased the left ventricular ejection fraction(LVEF) and left ventricular fractional shortening(LVFS), effectively improved cardiac function in mice after myocardial infarction, and effectively reduced the levels of myocardial injury markers such as creatine kinase(CK), creatine kinase isoenzyme(CK-MB), and lactic dehydrogenase(LDH), thus protecting ischemic myocardium. HE staining showed that Jiming Powder could attenuate myocardial inflammatory cell infiltration after myocardial infarction. Masson and Sirius Red staining demonstrated that Jiming Powder effectively inhibited myocardial fibrosis, reduced the collagen Ⅰ/Ⅲ ratio in myocardial tissues, and improved collagen remodeling after myocardial infarction. Western blot results showed that Jiming Powder reduced the expression of TGF-β1, α-SMA, Wnt3a, and β-catenin, decreased the levels of MMP2, MMP3, and TIMP2, and increased the level of TIMP1, suggesting its role in inhibiting cardiac fibroblast transformation, reducing extracellular matrix metabolism in myocardial cells, and lowering collagen Ⅰ and α-SMA content, thus exerting an anti-myocardial fibrosis effect after myocardial infarction. This study revealed the role of Jiming Powder in improving ventricular remodeling and treating myocardial infarction, laying the foundation for further research on the pharmacological effect of Jiming Powder.
Mice ; Animals ; Transforming Growth Factor beta1/metabolism* ; Matrix Metalloproteinase 2/metabolism* ; beta Catenin/metabolism* ; Matrix Metalloproteinase 3/therapeutic use* ; Powders ; Ventricular Remodeling ; Stroke Volume ; Ventricular Function, Left ; Myocardial Infarction/drug therapy* ; Myocardium/pathology* ; Heart Failure/metabolism* ; Collagen/metabolism* ; Creatine Kinase ; Fibrosis

BACKGROUND: Colorectal cancer is the third most common cancer worldwide and still lack of effective therapy so far. Petasin, a natural product found in plants of the genus Petasites, has been reported to possess anticancer activity. The present study aimed to investigate the anticolon cancer activity of petasin both in vitro and in vivo. The molecular mechanism of petasin was also further explored. METHODS: Caco-2, LoVo, SW-620, and HT-29 cell lines were used to detect the inhibitory effect of petasin on colon cancer proliferation. Cell viability was determined using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Cell apoptosis was analyzed by flow cytometry. Hoechst 33258 staining was used to visualize morphological changes. Cell migration was assessed using a wound-healing migration assay, and cell invasion was investigated using Transwell chambers. Western blotting assays were employed to evaluate the expression levels of proteins in the protein kinase B/mammalian target of rapamycin (Akt/mTOR) signaling pathway. Finally, in vivo activity of petasin was evaluated using the SW-620 subcutaneous tumor model established in Balb/c nude mice. Twelve rats were randomly divided into control group and 10 mg/kg petasin group. The tumor volume was calculated every 7 days for 28 days. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was performed to assess the apoptotic effect of petasin. Differences between two groups were assessed by analysis of independent-sample t tests. RESULTS: Petasin significantly inhibited the proliferation of human colon carcinoma cell lines, induced apoptosis, and suppressed migration and invasion in SW-620 cells. Western blotting results showed that petasin decreased the phosphorylation of Akt (1.01 ± 0.16 vs. 0.74 ± 0.06, P = 0.042), mTOR (0.71 ± 0.12 vs. 0.32 ± 0.11, P = 0.013), and P70S6K (1.23 ± 0.21 vs. 0.85 ± 0.14, P = 0.008), elevated the expression of caspase-3 (0.41 ± 0.09 vs. 0.74 ± 0.12, P = 0.018) and caspase-9 (1.10 ± 0.27 vs. 1.98 ± 0.22, P = 0.009), decreased the Bcl-2 protein (2.75 ± 0.47 vs. 1.51 ± 0.36, P = 0.008), downregulated the expression of matrix metalloproteinase (MMP)-3 (1.51 ± 0.31 vs. 0.82 ± 0.11, P = 0.021) and MMP-9 (1.56 ± 0.32 vs. 0.94 ± 0.15, P = 0.039) in SW-620 cell. In vivo, 10 mg/kg petasin inhibited tumor growth in Balb/c nude mice (924.18 ± 101.23 vs. 577.67 ± 75.12 mm at day 28, P = 0.001) and induced apoptosis (3.6 ± 0.7% vs. 36.0 ± 4.9%, P = 0.001) in tumor tissues. CONCLUSIONS Petasin inhibits the proliferation of colon cancer SW-620 cells via inactivating the Akt/mTOR pathway. Our findings suggest petasin as a potential candidate for colon cancer therapy.
Animals ; Antineoplastic Agents ; therapeutic use ; Apoptosis ; drug effects ; Caco-2 Cells ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; HT29 Cells ; Humans ; In Situ Nick-End Labeling ; Matrix Metalloproteinase 3 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Phosphorylation ; drug effects ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Sesquiterpenes ; therapeutic use ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; genetics ; metabolism

OBJECTIVETo study the effect of Shengji Huayu Recipe (SHR)on the expression of MMP-3 and TIMP-1 in the skin ulcer tissue of diabetic rats.

METHODSThe skin ulcer model was established in diabetic mice. Different compatibility proportions of SHR [the ratio of Shengji Recipe (SJR) to Huayu Recipe (HYR) = 2:1, 1:1, and 1:2, respectively] were used to intervene. The expression of MMP-3 protein in the skin ulcer of diabetic rats was detected by Western blot method,and TIMP-1 protein was detected by immunohistochemical assay.

RESULTSAt each time point, there was no statistical difference in the blood glucose level among groups (P > 0.05). But all of them increased significantly,when compared with those of the normal wound group (P < 0.01). As for the difference between after would area treatment and before would area treatment, better effect was obtained in the SHR No. 3 group and the normal ulcer group than in the diabetic ulcer model group (P < 0.05). Results of Western blot showed that the MMP-3 protein expression was higher in the SHR No. 2 group than in the SHR No.3 group (P < 0.05). Immunohistochemical results showed that TIMP-1 protein expression was lower in the SHR No. 2 group than in the SHR No. 3 group and the diabetic ulcer model group (P < 0.05). TIMP-1 protein expression was higherin the SHR No. 3 group than in the SHR No. 2 group (P < 0.01).

CONCLUSIONUsing SHR No.3 was conducive to the promotion of wound healing in early wound repair stage, and using SHR No. 2 might be conducive to inhibiting the formation of pathological scar.

Animals ; Diabetes Mellitus, Experimental ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Male ; Matrix Metalloproteinase 3 ; metabolism ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Skin ; drug effects ; pathology ; Skin Ulcer ; drug therapy ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism

OBJECTIVETo investigate the anti-inflammatory effect of Herba Siegesbeckiae extracts on mouse rheumatoid arthritis induced by arthrogen-CIA monoclonal antibody.

METHODSThe rheumatoid arthritis was induced by arthrogen-CIA arthritogenic monoclonal antibody in mice. The sandwich enzyme-linked immunosorbent assay was used to determine the concentration of IL-1βin mouse serum,and the content of IL-6,IL-17 and MMP-3 in supernatant of tissue homogenate of hind limb below the stifle of mice. One-way ANOVA was used for data analysis.

RESULTSThe toe swelling was attenuated in Siegesbeckiae group than that in model group [(0.218 ± 0.0307)cm(3) compared with (0.2545 ± 0.0179)cm(3), P<0.05]. The serum IL-1β level in Siegesbeckiae group was lower than that in model group [(63.74 ± 21.74)pg/ml compared with (104.96 ± 31.22)pg/ml, P<0.01]. The contents of IL-6, IL-17 and MMP-3 in tissue supernatants of Siegesbeckiae group were all lower than those of model group [(171.10 ± 48.35)pg/ml compared with (249.64 ± 75.08)pg/ml, P<0.05; (115.42 ± 56.52)pg/ml compared with (208.40 ± 88.54)pg/ml, P<0.05;(3660.31 ± 1680.99) pg/ml compared with (5420.79 ± 1201.43)pg/ml, P<0.05, respectively].

CONCLUSIONThe extract of Herba Siegesbeckiae has anti-inflammatory effect on mouse rheumatoid arthritis induced by mixed arthrogen monoclonal antibody.

Animals ; Anti-Inflammatory Agents ; therapeutic use ; Antibodies, Monoclonal ; Arthritis, Experimental ; drug therapy ; Arthritis, Rheumatoid ; chemically induced ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Interleukin-17 ; metabolism ; Interleukin-1beta ; blood ; Interleukin-6 ; metabolism ; Matrix Metalloproteinase 3 ; metabolism ; Mice ; Mice, Inbred BALB C

This study was conducted to determine if nasal salmon calcitonin has additional beneficial effects on clinical symptoms, serum NO, IL-1beta, matrix metalloproteinase 3, urinary C-terminal telopeptide type II collagen (CTX-II) levels and MRI findings in knee osteoarthritis (OA) when used concomitantly with exercise therapy. Fifty female patients with knee OA were randomized into two groups. The first group (n = 30) received 200 IU/day nasal salmon calcitonin and a home exercise program; the second group (n = 20) received a home exercise program for 6 months. Compared with baseline,while significant improvements were observed in visual analogue scale (VAS), WOMAC pain, physical function scores, 20-m walking time (P < 0.001) and WOMAC stiffness score (P = 0.041) in the first group, walking and resting VAS, and WOMAC physical function scores were improved (P = 0.029) in the second group after treatment. Significantly increased levels of serum NO and urinary CTX-II (P < 0.001) and significant improvements in the area of medial femoral condyle (P < 0.05) were noted only in the first group. There were significant differences in VAS activation values (P = 0.032) and NO levels (P < 0.001) in the favor of the first group. In conclusion, nasal salmon calcitonin may have possible chondroprotective effects besides its known effects on symptoms in patients with knee OA.
Aged ; Bone Density Conservation Agents/*therapeutic use ; Calcitonin/*therapeutic use ; Collagen Type II/urine ; Exercise Therapy ; Female ; Humans ; Interleukin-1beta/blood ; Magnetic Resonance Imaging ; Matrix Metalloproteinase 3/blood ; Middle Aged ; Nitric Oxide/blood ; Osteoarthritis, Knee/*drug therapy/radiography ; Peptide Fragments/urine ; Respiratory Therapy ; Severity of Illness Index ; Treatment Outcome ; Walking

This study was conducted to determine if nasal salmon calcitonin has additional beneficial effects on clinical symptoms, serum NO, IL-1beta, matrix metalloproteinase 3, urinary C-terminal telopeptide type II collagen (CTX-II) levels and MRI findings in knee osteoarthritis (OA) when used concomitantly with exercise therapy. Fifty female patients with knee OA were randomized into two groups. The first group (n = 30) received 200 IU/day nasal salmon calcitonin and a home exercise program; the second group (n = 20) received a home exercise program for 6 months. Compared with baseline,while significant improvements were observed in visual analogue scale (VAS), WOMAC pain, physical function scores, 20-m walking time (P < 0.001) and WOMAC stiffness score (P = 0.041) in the first group, walking and resting VAS, and WOMAC physical function scores were improved (P = 0.029) in the second group after treatment. Significantly increased levels of serum NO and urinary CTX-II (P < 0.001) and significant improvements in the area of medial femoral condyle (P < 0.05) were noted only in the first group. There were significant differences in VAS activation values (P = 0.032) and NO levels (P < 0.001) in the favor of the first group. In conclusion, nasal salmon calcitonin may have possible chondroprotective effects besides its known effects on symptoms in patients with knee OA.
Aged ; Bone Density Conservation Agents/*therapeutic use ; Calcitonin/*therapeutic use ; Collagen Type II/urine ; Exercise Therapy ; Female ; Humans ; Interleukin-1beta/blood ; Magnetic Resonance Imaging ; Matrix Metalloproteinase 3/blood ; Middle Aged ; Nitric Oxide/blood ; Osteoarthritis, Knee/*drug therapy/radiography ; Peptide Fragments/urine ; Respiratory Therapy ; Severity of Illness Index ; Treatment Outcome ; Walking

OBJECTIVETo investigate the value of metalloproteinase-3 (MMP-3) levels in assessing efficacy of etanercept treatment in patients with ankylosing spondylitis (AS).

METHODSThe serum and synovial fluid levels of MMP-3 were measured by enzyme linked immunosorbent assay (ELISA) in 48 patients with AS in week 0, 6 and 12; and also measured in 30 serum samples and 10 synovial fluid samples from healthy controls.

RESULTSThe serum levels of MMP-3 in AS patients were significantly higher than those in controls. In AS patients, the MMP-3 levels in synovial fluid were significantly higher than those in serum levels. The serum MMP-3 levels in AS patients with peripheral arthritis were higher than those with exclusively axial involvement; while C-reactive protein (CRP) levels and erythrocyte sedimentation rate (ESR) did not differ between these 2 groups of AS. At week 6 and week 12 of etanercept treatment, the serum MMP-3 levels were significantly decreased (p<0.01) with the declining trend of ESR, CRP, Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and Bath Ankylosing Spondylitis Functional Index (BASFI) (all p<0.01). Before the etanercept treatment (week 0), serum levels of MMP-3 were correlated with ESR, CRP, BASDAI and BASFI (p<0.05). ESR was also correlated with CRP and BASFI, but not with BASDAI (r=0.361, P=0.071). At weeks 12, serum MMP-3 levels were still correlated with ESR, CRP and BASDAI (P<0.05), but not with BASFI (P=0.339); ESR was correlated with CRP, but not with BASDAI and BASFI. There was a significant correlation between BASDAI and BASFI (r=0.818,P=0.001).

CONCLUSIONSerum MMP-3 levels are closely related to disease activity and may serve as an useful indicator for efficacy of etanercept treatment in AS patients.

Adolescent ; Adult ; Aged ; Antirheumatic Agents ; therapeutic use ; Biomarkers ; blood ; Etanercept ; Female ; Humans ; Immunoglobulin G ; therapeutic use ; Male ; Matrix Metalloproteinase 3 ; blood ; Middle Aged ; Receptors, Tumor Necrosis Factor ; therapeutic use ; Spondylitis, Ankylosing ; blood ; drug therapy ; Young Adult

OBJECTIVETo evaluate the efficacy and safety of etanercept, a tumor necrosis factor (TNF)-alpha inhibitor, in the treatment of ankylosing spondylitis (AS), and investigate its effect on serum levels of matrix metalloproteinase-3 (MMP-3).

METHODSForty-eight patients with AS received etanercept 25 mg twice a week for a treatment course of 12 weeks. The patients' symptoms, signs, Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), Bath Ankylosing Spondylitis Functional Index (BASFI), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) levels and side effects were observed before and after the treatment. The serum levels of MMP-3 was determined using enzyme-linked immunosorbent assay (ELISA).

RESULTSAll the patients completed the treatment. The degree of spinal pain and pain at night, the duration of morning stiffness, the finger-to-floor distance, BASDAI and BASFI were significantly improved after the treatment (P<0.05). Etanercept treatment resulted in a significant reduction in serum MMP-3 level in the AS patients to 31.22-/+10.26 ng/ml as compared with the level before treatment (46.17-/+25.74 ng/ml, P<0.05). The reduction of serum MMP-3 was positively correlated to decrement of ESR and CRP (r=0.397 and 0.474, respectively, P<0.05). The most common adverse events of etanercept included injection site reaction and upper respiratory infection.

CONCLUSIONEtanercept treatment has obvious therapeutic effects on AS without serious adverse effects. MMP-3 may be a potentially useful indicator to assess the effect of anti-TNF-alpha treatment in AS patients.

Adolescent ; Adult ; Antirheumatic Agents ; therapeutic use ; C-Reactive Protein ; metabolism ; Etanercept ; Female ; Humans ; Immunoglobulin G ; therapeutic use ; Male ; Matrix Metalloproteinase 3 ; blood ; Middle Aged ; Receptors, Tumor Necrosis Factor ; therapeutic use ; Spondylitis, Ankylosing ; blood ; drug therapy ; pathology ; Treatment Outcome ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors ; Young Adult

OBJECTIVE: To examine the expression of MMP-3 and TIMP-1 in the synovial fluid in knee joints with osteoarthritis before and after being treated with hyaluronic acid(HA), glucosamine sulfate(GS) and arthroscopic de bridment(AD), and to explore the therapeutic mechanism. METHODS: Sixty patients (64 knees) with osteoarthritis(OA) were randomly divided into HA group AD group and GS+AD group. Some patients from the HA group and the GS+HA group were selected, and served as HA' group and GS+HA' group. The expression of MMP-3 and TIMP-1 in the synovial fluid was measured by enzyme-linked immunosorbent assay (ELISA) before and after 4 week and 6 month therapy. RESULTS: The level of MMP-3 and the ratio of MMP-3/TIMP-1 in the synovial fluid decreased after being treated for 4 weeks, and the effect on MMP-3 and MMP-3/TIMP-1 lasted for 6 months in the HA group and the GS+HA groups. The levels of TIMP-1 increased significantly after being treated for 4 weeks only in the GS+HA group. The level of MMP-3 and the ratio of MMP-3/TIMP-1 in the synovial fluid decreased, but the level of TIMP-1 increased after being treated for 4 weeks, and the effect on MMP-3 and MMP-3/TIMP-1 lasted for 6 months. The level of MMP-3 and the ratio of MMP-3/TIMP-1 in the synovial fluid increased after being treated for 6 months compared with those for 4 weeks. The level of TIMP-1 increased in the GS group more than that in the HA group after being treated for 4 weeks. The level of TIMP-1 increased in the AD group more than that in the HA' group and the GS+HA' group after being treated for 4 weeks. CONCLUSION (1) HA, GS and AD all can decrease the level of of MMP-3 and the ratio of MMP-3/TIMP-1 in the synovial fluid in knee joints with OA. (2) The level of TIMP-1 in the synovial fluid has no difference before and after being treated with HA. The AD group and GS group can increase the level of TIMP-1, which indicates that the AD group or GS group might produce better therapeutic effect. (3) The level of TIMP-1 increased in the AD group is more than that in the HA' group and the GS+HA' group after being treated for 4 weeks, which indicates that the AD group might get better therapeutic effect than the other 2 groups. (4) One of the most important mechanisms of HA, GS and AD in treating OA might be attributed to the expression of MMPs and TIMPs in knee joints with OA.
Aged ; Debridement ; methods ; Female ; Glucosamine ; therapeutic use ; Humans ; Hyaluronic Acid ; therapeutic use ; Male ; Matrix Metalloproteinase 3 ; metabolism ; Middle Aged ; Osteoarthritis, Knee ; metabolism ; therapy ; Synovial Membrane ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism

OBJECTIVETo investigate the mechanism of Tougu Xiaotong Granula (TXG) on prevention and treatment of knee osteoarthritis.

METHODSFifty New Zealand rabbits were randomly divided into 6 groups. Except those in the normal control group, all the rabbits were replicated into knee osteoarthritis model using modified Hulth method. They were administered by gastrogavage once every day respectively with 100 ml of normal saline to the rabbits in the normal group and those in the model group, with 10 g of Zhuanggu Guanjie Pill to those in the control group, and 5 g, 10 g and 20 g of TXG to those in the three TXG tested groups (tested group 1, 2 and 3). The levels of interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and metalloproteinase 3 (MMP-3) in joint fluid, the blood content of malondialdehyde (MDA) and nitric oxide (NO) absorbance as well as the SOD activity in synovia were observed.

RESULTSOverexpressions of IL-1, IL-6, TNF-alpha and MMP-3 in joint fluid, increased blood content of NO and MDA were shown in the 8th and 16th week, and decreased SOD activity in synovia was shown in the 16th week of the experiment in all the model rabbits, as compared to those in the normal group, the difference was significant respectively (P < 0.05 or P<0.01). After 8 weeks of treatment, the levels of IL-1, TNF-alpha, MMP-3, NO and MDA in the control group, tested group 2 and 3 were significantly different to those in the model group respectively (P < 0.05, P < 0.01), and significant difference was also shown in the comparisons of those indexes between the control group and the tested group 1 vs the tested group 3 (P < 0.05). As for the level of IL-6, significant difference was shown in comparisons of the model group with the control group, tested group 2 and 3 in the 8th and 16th week of the treatment (P < 0.05 or P < 0.01), also in comparison of the tested group 3 with the tested group 1 in the 8th week, and in that of the tested group 2 with the control group and the tested group 1 in the 16th week (P < 0.05).

CONCLUSIONTXG could effectively postpone the degeneration of cartilage through effectively inhibiting the biological effects of cytokines, MMP-3 and oxygen free radical.

Animals ; Cytokines ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Female ; Male ; Malondialdehyde ; blood ; Matrix Metalloproteinase 3 ; metabolism ; Nitric Oxide ; blood ; Osteoarthritis, Knee ; blood ; drug therapy ; metabolism ; Phytotherapy ; Rabbits ; Superoxide Dismutase ; metabolism