Maxing xiongting mixture regulates factors relevant to lung reshaping and vascular remodeling of hypoxic pulmonary hypertension rats / 中国组织工程研究

BACKGROUND: Hypoxic pulmonary hypertension is a key link in the progression from chronic obstructive pulmonary disease to cor pulmonale. Its severity is closely related to disease development and prognosis. Current treatments cannot prevent or reverse disease progression. Maxing Xiongting Mixture has significant effect on hypoxic pulmonary hypertension with the syndrome of intermingled phlegm and blood stasis. OBJECTIVE: To study how the Maxing Xiongting Mixture regulates relevant factors of lung reshaping and vascular remodeling of hypoxic pulmonary hypertension rats with the syndrome of intermingled phlegm and blood stasis. METHODS: Seventy Sprague-Dawley rats, 5 weeks old, were randomly divided into normal group (n=10) and model group (n=60), where acute cor pulmonale model was prepared by injecting 50 mg/kg monocrotaline solution (1%) intraperitoneally, followed by forced smoking and swimming 6 days a week lasting for 4 weeks. Except for 10 rats in the normal group, there were 46 model rats in the model group. According to the normal distribution of body mass, 40 rats were selected and randomly divided into 4 groups: model group, high-dose Maxing Xiongting Mixture group (MH), low-dose Maxing Xiongting Mixture group (ML) and fasudil group, with 10 rats in each group. Rats in MH and ML groups were respectively given Maxing Xiongting Mixture at 20 g/(kg·d) and 5 g/(kg·d), respectively and those in the fasudil group were given fasudil at a dose of 10 mg/(kg·d). Other groups were given equal amount of saline. Administration was given intraperitoneally and intragastrically, once a day for 14 days in total. RT-PCR was used to test the expression of factors related to lung reshaping and vascular remodeling, including RhoA, stromelysin 1 and tumor necrosis factor-α mRNAs. An approval for the study was obtained from the Ethics Committee of Chengdu University of Traditional Chinese Medicine (approval No. 2017-03). RESULTS AND CONCLUSION: Compared with the model group, the expressions of RhoA, stromelysin 1, and tumor necrosis factor-α mRNAs were significantly lowered in the MH group (all P 0.05). To conclude, Maxing Xiongting Mixture, which is similar to fasudil, intervenes lung reshaping and vascular remodeling of hypoxic pulmonary hypertension rats with the syndrome of intermingled phlegm and blood stasis by inhibiting the expressions of RhoA, stromelysin 1, and tumor necrosis factor-α mRNAs.

Heterozygosity of stromelysin‑1 (rs3025058) promoter polymorphism is associated with gastric cancer.

BACKGROUND: Gastric cancer (GC) is the third most common cancer in India and is mediated by multiple genetic, epigenetic and environmental risk factors. A single nucleotide polymorphism rs3025058 at − 1171 of the stromelysin‑1 (matrix metalloproteinase [MMP]‑3) promoter is resulting due to insertion/deletion of adenine thought to have an impact on increasing the risk for tumor formation. AIM: This study is aimed to understand the role of stromelysin‑1 rs3025058 (−1171, 5A/6A) promoter polymorphism in the etiology of GC in Indian population. MATERIALS AND METHODS: Genomic DNA was isolated from blood samples of the GC patients and controls. The genotyping of stromelysin‑1 rs3025058 (−1171, 5A/6A) promoter polymorphism was carried out by amplification refractory mutation system‑polymerase chain reaction method followed by agarose gel electrophoresis. RESULTS: The frequency of 5A/5A, 5A/6A, and 6A/6A genotypes in GC patients were 7.69%, 76.92%, and 15.38%, while in controls were 5.31%, 86.73%, and7.96%, respectively. There was a significant difference in the distribution of 5A/6A genotype in patients compared to the controls (P < 0.05). CONCLUSION: This study showed an increased frequency of heterozygotes for stromelysin‑1 rs3025058 and thought to be involved in the etiology of GC.

Changes of matrix metalloproteinase and cytokines and its clinical significance in children with juvenile idiopathic arthritis / 中华风湿病学杂志

Objective The purpose of the study was to investigate the changes and clinical significance of matrix metalloproteinase (MMP)-1,MMP-3,tissue inhibitor of metalloproteinase (TIMP),interleukin (IL)-1,tumor necrosis factor (TNF)-α and transforming growth factor(TGF)-β in juvenile idiopathic arthritis (JIA).Methods Thirty-two JIA subjects and 28 controls (traumatic arthritis patients) were included into this study.The MMP-1,MMP-3,TIMP,IL-1,TNF-α and TGF-β level in the serum and synovia were assessed by ELISA.The WBC count,the level of CRP,ESR,RF were also detected.Independent t-test and Pearson's analysis were adopted for data analysis.Results ① The level of MMP-1,MMP-3,IL-1 and TNF-α in the serum was (158±67) ng/ml,(212±89) ng/ml,(39±19) pg/ml,(26±10) pg/ml respectively,which was significantly higher than that of the control group (all P＜0.05) ; the ratio of MMP-3/TIMP-1 (0.86±0.32) was higher in the study group than that of the control group (P＜0.05),while the value of TIMP-1,TGF-β was (248±88),(17±9) ng/ml respectively,which was lower than that of the control group (P＜0.05).The value of seral MMP-3,MMP-3/TIMP-1 was positively correlated with that of WBC,CRP,ESR in the JIA group (all P＜0.05).② The value of MMP-1,MMP-3,IL-1 and TNF-α in the synovia was (216±78) ng/nl,(766±291)ng/ml,(56±21) pg/ml,(36±14) pg/ml respectively,which was higher than that of the control group (all P＜0.05); the ratio of MMP-3/TlMP-1 (2.68±0.89) was higher than that of the control group (P＜0.05),while the value of TIMP-1,TGF-β was (286±88) ng/ml,(12±4) ng/ml respectively,which was lower than that of the control group (all P＜0.05).③ The value of MMP-1,MMP-3,TIMP-1,IL-1 and TNF-α in the synovia was higher than that in the serum (all P＜0.05),while the value of TGF-β was lower than that in the serum (P＜0.05).Conclusion The value of MMP-1,MMP-3,IL-1 and TNF-α increases both in the serum and synovia,while the value of TIMP-1 decreases.The value of TGF-β decreases,which may have protective effect on JIA.The ratio of MMP-3/TIMP-1 in the serum is positively correlated to inflammation parameters,which may be used to judge the activity of illness in JIA.

Morphological studies on osteoarthritis of the articular cartilage in C57 black mice and its relationship with etalloproteinase-3 and lnterleukln-1 in serum and synovial fluid / 中华风湿病学杂志

Objective To study the levels of Matrix Metalloproteinase-3 (MMP-3) and interleukin (IL-1) in the synovial fluid and plasma of C57 black mice with osteoarthritis (OA) and their relationships with the severity of pathological changes so as to investigate their effects and correlation with OA. Methods The C57 black mice with OA were enrolled for this study. Different levels of exercise were appicated based on their age. Knee joint pathological changes were examined for pathological severity of OA. ELISA sandwich method was used to measure the levels of MMP-3 and IL-1 in serum and synovial fluid. Correlation analysis was performed to demonstrate the relationship between the levels of MMP-3 and IL-1 in the serum and synovial fluid, and the pathological severity of OA. Results ①Morphological observations: C57 black mice were characterized by spontaneously developing OA and the incidence and the severity of osteoarthritis gradually increased with age and exercising burdens. ② The level of MMP-3 and IL-1 in the synovial fluid of exercising mice MMP-3 (84±6) ng/ml, IL-1 (48±3) ng/ml was higher than that in the aged ones [MMP-3 (84±6) ng/ml, IL-1 (71±5) ng/ml J, the difference was significant (P<0.01). The level of MMP-3 and IL-1 level in the serum had a linear correlation with that of the synovial fluid. At the same time, they also had linear correlation with the pathological severity of OA (All r>0.67, and all P<0.01). Conclusion The levels of MMP-3 and IL-1 in serum and synovial fluid can help to make early diagnosis of OA, especially elevated MMP-3 level.

Stromelysin-1 and Membrane type-MMP-1 Expressions in Human Chronic Periodontitis with Type 2 Diabetes Mellitus / 대한치주과학회지

PURPOSE: The purposes of this study were to compare and quantify the expression of Stromelysin-1 and MT-MMP-1 in the gingival tissues of patients with type 2 diabetes mellitus(DM) and healthy adults with chronic periodontitis. MATERIALS AND METHODS: Gingival tissue samples were obtained during periodontal surgery or tooth extraction. According to the patient's systemic condition & clinical criteria of gingiva, each gingival sample was devided into three groups. Group 1 (n=8) is clinically healthy gingiva without bleeding and no evidence of bone resorption or periodontal pockets, obtained from systemically healthy 8 patients. Group 2 (n=8) is inflammed gingiva from patients with chronic periodontitis. Group 3 (n=8) is inflammed gingiva from patients with chronic periodontitis associated with type 2 DM. Tissue samples were prepared and analyzed by Western blotting. The quantification of Stromelysin-1 and MT-MMP-1 were performed using a densitometer and statistically analyzed by one-way ANOVA followed by Tukey test. RESULTS: In the analysis of expression levels, Stromelysin-1 and MT-MMP-1 expressions were similar in group 1 and 2. Stromelysin-1 and MT-MMP-1 expressions was more increased in group 3 than group 1, 2. The difference between group 3 and group 1, 2 was statistically significant. Also, in the interrelationship of Stromelysin-1 and MT-MMP-1 expressions, expressions of Stromelysin-1 and MT-MMP-1 showed increasing tendency in chronic periodontitis associated with type 2 DM and it seems that the MT-MMP-1 expressions were increasing in proportion to Stromelysin-1 expressions. CONCLUSION: It is suggested that Stromelysin-1 and MT-MMP-1 may be partly involved in the progression of periodontal inflammation associated with type 2 DM, as related to a metabolism of other factors, such as AGE, plasmin and other inflammatory mediators. Therefore, the expression levels of Stromelysin-1 and MT-MMP-1 can be inflammatory markers of periodontal inflammed tissue with type 2 DM.

Microglia, major player in the brain inflammation: their roles in the pathogenesis of Parkinson's disease

Inflammation, a self-defensive reaction against various pathogenic stimuli, may become harmful self-damaging process. Increasing evidence has linked chronic inflammation to a number of neurodegenerative disorders including Alzheimer's disease (AD), Parkinson's disease (PD), and multiple sclerosis. In the central nervous system, microglia, the resident innate immune cells play major role in the inflammatory process. Although they form the first line of defense for the neural parenchyma, uncontrolled activation of microglia may directly toxic to neurons by releasing various substances such as inflammatory cytokines (IL-1beta, TNF-alpha, IL-6), NO, PGE

The Roles of Stromelysin-1 and the Gelatinase B Gene Polymorphism in Stable Angina

Matrix metalloproteinases contribute to vascular remodeling by breaking down extracellular-matrix while new matrix is synthesized. Of the variety of MMPs, stromelysin-1 and gelatinase B may have key roles in coronary artery atherosclerosis. Moreover, The 5A/6A polymorphism in the promoter region of the stromelysin-1 gene may be a pathogenetic risk factor for acute myocardial infarction. Gelatinase B (92-kDa type IV collagenase and MMP-9) is one of the MMPs found to be highly expressed in the disruption-prone regions of atherosclerotic plaques. C- to T substitution at the promoter site (-1562) resulted in the higher promoter activity of the T-allelic promoter. The R279Q polymorphism in exon 6 led to the substitution of adenosine by guanine, and was a common polymorphism in the general population. We evaluated the relation between these polymorphisms and stable angina, the severity of atherosclerosis in coronary artery disease, and instent restenosis after percutaneous coronary angioplasty. The study population was composed of 131 patients with stable angina (mean age 61.3 years, 89 males) and 117 control subjects (mean age 59.3 years, 59 males). Coronary angiographies were performed in all cases at Yonsei University Cardiovascular Hospital from February 1998 to June 2000. The genotype for each polymorphism was determined using a SNaPshotTM kit and by restriction fragment length polymorphism (RFLP). The prevalence of 5A containing a polymorphism of the stromelysin-1 gene was higher in the stable angina group than in control patients, but no difference in the two polymorphisms of the gelatinase B gene was found between the two groups. By multiple logistic analysis, the 5A-allele of the stromelysin-1 gene was found to be an independent risk factor of stable angina with an odds ratio of 2.29 (95% CI; 1.19-4.38). However, the severity of atherosclerosis in coronary artery or in stent restenosis was not related to any polymorphism of stromelysin-1 or gelatinase B. Our results show that functional genetic variation of stromelysin-1 could be a significant risk factor for stable angina, and might play an important role in coronary atherosclerosis involving vascular remodeling.

Effects of matrix metalloproteinase 3 on wound healing: an experimental study / 解放军医学杂志

Objective To explore the changes and significance of matrix metalloproteinase 3 (MMP3) during wound healing in rat. Methods Wound fluids and dermal pluripotent stem cells (DPSCs) were isolated with routine method from rats, and they were purified and expanded. Wound fluid was collected on the first day to serve as wound microenvironment, then the responses of DPSCs to wound fluids were investigated, and the expression of MMP3 protein in DPSCs was determined by immunohistochemistry staining. Meanwhile, animal models were repruduced with cutaneous incision and suturing, and the animals were respectively assigned to intractable wound group, in which rats received whole body irradiation, and simple wound group, in which no irradiation was given. The rats were sacrificed on 3rd, 5th, 7th, 10th and 14th day posttrauma (n=5 each), and specimens of wound tissue were harvested, fixed with 10% formalin solution. Twenty four hours later, the samples were dehydrated and embeded, then paraffin section were made. Paraffin sections were stained with hematoxylin and eosin (HE) staining. Light microscopy was used to observe the pathological changes in wounds. Five randomly selected fields were observed under a ?40 objective to evaluate the histological features, especially the amount of tissue repairing cells in wounds. Furthermore, MMP3 contents in wound sites were determined by immunohistochemistry assay and image analysis. Results The expression of MMP3 in DPSCs increased significantly after stimulation by wound fluids. MMP3 contents in rats of simple wound group increased significantly, especially in the dermal tissues, and the peak value appeared 5-7 days after trauma. MMP3 contents in rats of intractable wound group were significantly less than those of simple wound group, and the time when the peak value appeared was also delayed till the 10th day after trauma. Conclusions MMP3 may be an important substrate involved in wound healing. DPSCs may participate in the processes of wound repairing via high expression of MMP3.

EFFECTS OF RECOMBINANT HUMAN AUGMENTER OF LIVER REGENERATION ON STROMELYSIN-1 GENE EXPRESSION IN RATS WITH LIVER FIBROSIS / 解放军医学杂志

To investigate the influence of recombinant human augmenter of liver regeneration (hALR) on stromelysin 1 gene expression in rats with fibrotic liver. CCl 4 or albumin induced liver fibrosis in rats was established, and different dosages of recombinant human augmenter of liver regeneration were given to rats with liver fibrosis.Liver specimens were obtained at different intervals of treatment , total RNA of liver tissues were isolated and stromelysin 1 gene expression was measured by reverse transcription polymerase chain reaction (RT PCR) . The results showed that in both rat models of experimental liver fibrosis , stromelysin 1 gene expression levels were significantly higher in hALR treated rats than those without the treatment at various intervals. Stromelysin 1 gene expression levels in high dose hALR treatment group were significantly higher than that in low dose hALR treatment group. It suggested that recombinant human augmenter of liver regeneration may enhance stromelysin 1 gene expression in rats with fibrotic liver.

Recombinant human augmenter of liver regeneration may promote stromelysin-1 gene expression in experimental liver fibrosis / 中华传染病杂志

Objective To investigate the influence of recombinant human augmenter of liver rege neration (hALR) on stromelysin 1 gene expression in experimental liver fibrosis. Methods Two kinds of rat model of experimental liver fibrosis induced by CCl 4 and albumin were established and different dosages of recombinant human augmenter of liver regeneration were given during the process of model making. Total RNA of liver tissues was extracted and stromelysin 1 gene expression levels were measured by reverse transcription polymerase chain reaction (RT PCR). Results In both rat models of experimental liver fibrosis, stromelysin 1 gene expression levels in hALR treating group were significantly higher than those of model groups in different periods of model forming. Stromelysin 1 gene expression levels in high dose hALR treated group were significantly higher than those of low dose hALR treated group.Conclusion Recombinant human augmenter of liver regeneration may have effects of promoting gene expression of stromelysin 1 in experimental liver fibrosis.

Expression of matrix metalloproteinase-2,9 and it′s tissue inhibitor-1,2 in endometrial carcinoma / 中华妇产科杂志

Objective To study the expression of matrix metalloproteinase(MMP)-2,9 and tissue inhibitor of metalloproteinase(TIMP)-1,2 protein in human endometrial carcinoma tissue and its relation to the invasion and metastasis of endometrial carcinoma. Methods Immunocytochemistry and zymography techniques were used to measure the MMP-2,MMP-9,TIMP-1,TIMP-2 protein levels and activities in endometrial carcinoma tissue of 37 patients and control group composed of 7 normal postmenstrual endometrial samples. Results The MMP-2,MMP-9,TIMP-1 and TIMP-2 proteins mainly expressed in endometrial carcinoma cells, glandular cells and endothelial cells. The strongly positive expression proportions of MMP-2,9 and TIMP-1 proteins in grade Ⅲ carcinoma cells were respectively 73%, 20% and 67%, which were higher than those in gradeⅡ (13%, 0, 27%) and gradeⅠ (0) ones ( P

Effect of matrix metalloproteinase-3 on vitreous and vitreoretinal interface in rabbit's eye / 中华眼底病杂志

Objective To investigate the effects of intravitreal injection of matrix metalloproteinase-3 (MMP-3) on the vitreoretinal adhesion and the vitreous gelatin. Methods Twenty-four pigmented rabbits were randomly divided into 3 experimental groups (group A, B, and C)and one control group with 6 rabbits (12 eyes) in each. Different concentrations of 0.1 ml MMP-3 (5,10, 20 ng in group A, B, and C, respectively) and equivalent dose of balanced salt solution were intravitreally injected to the rabbits, respectively. Clinical examinations (such as gross observation, slit-lamp biomicroscopy, indirect fundus ophthalmoscopy ), electroretinography (ERG) and fundus fluorecein angiography (FFA) were taken before and after injection. Results One week after injection, posterior vitreous detachment (PVD) and focal vitreous liquefaction were recognized clinically for the first time in 1 eye in group B. By the end of this study, clinically detected PVD developed in 1 eye in group A, 3 eyes in group B, but the synchisis developed slowly, and no liquefaction or PVD occurred in control group. As for the histological examination, partial PVD was observed in 1 eye in group A and 3 eyes in group B 60 minutes after injection. All of the eyes in group A and B showed partial PVD 1 week after injection, and the area of PVD enlarged in contrast with before. Complete PVD were recognized in 1 eye in group A and 3 eyes in group B 15 weeks after injection, and the cleavage was narrow and limited. In group C, inflammatory cell infiltration in the inner layer of retina, destruction of retinal structure, and fluorescein leakage at late phase was found in the early period after injection. Conclusions MMP-3 is effective in disrupting the adhesion of the posterior hyaloid to the inner limiting membrane leading to PVD, and helpful to some extent in producing vitreous liquefaction. The dose of 10 ng MMP-3 is safe and effective for the rabbits eyes, which may be used as an promising assistant of vitreous surgery.

Interferon-gamma upregulates the stromelysin-1 gene expression by human skin fibroblasts in culture

The equilibrium between deposition and degradation of extracellular matrix(ECM) is essential to normal tissue development and repair of wound or inflammatory responses. It has recently become apparent that several cytokines and growth factors are capable of modulating fibroblast proliferation and biosynthetic activity. To understand the role of these factors in connective tissue regulation, we examined the effect of interferon-gamma (IFN-gamma) on stromelysin-1 gene expression in cultured human dermal fibroblasts. The steady-state levels of stromelysin-1 mRNA were increased in IFN-gamma treated cultured dermal fibroblasts. In the CAT assay, the stromelysin-1 promoter activity was increased 2.8-fold compared with untreated control. Therefore IFN-gamma stimulates the stromelysin-1 promoter activity, resulting in transcriptional enhancement of gene expression. Transforming growth factor-beta (TGF-beta) showed the antagonistic action to the effects of IFN-gamma in cultured dermal fibroblasts. Furthermore, gel mobility shift assays demonstrated enhanced AP-1 binding activities in nuclear extracts from cells incubated with IFN-gamma. These data suggest that IFN-gamma is an up-regulator and TGF-beta is a down regulator on the stromelysin-1 gene expression, respectively, and the AP-1 binding site may be necessary for gene response.

Interleukin-10 Differently Regulates Type I Collagen and Stromelysin-1 Promoter Activities in Dermal Fibroblast Cultures

BACKGROUND: Interleukin-10(IL-10) was initially discovered and isolated on the basis of its ability to suppress cytokine by Thl helper T cell. Recently, the effect of IL-10 has been reported in cultured connective tissue cells. OBJECTIVE: To further investigate the mechanisms of IL-10 on extracellular matrix(ECM) homeostasis, we evaluated the expression of type I collagen and stromelysin-1 at the transcriptional level and also observed their promoters activities. METHODS: We examined that effect of the recombinant human IL-10 on the expression of genes involved in extracelluar matrix(ECM) synthesis and remodelimg in human dermal fibroblast cultures. Quantification of collagen synthesis, Northern blot analysis, transient transfection and a chloramphenicol acetyl transferase(CAT) assay were performed. RESULTS: In studying the dose and time response effect of IL-10 on collagen synthesis, maximal reduction was seen at 1.0 ng/ml concentration and 24 hours incubation. The synthesis of type I collagen mRNA was downregulated, while stromelysin-1 gene expression was enhanced by IL-10. In the transient transfection and CAT assay, the type I collagen promoter/CAT reporter gene construct showed downregulation by IL-10, while the stromelysin-1 gene promoter activities were upregulated. CONCLUSION: It is suggested that IL-10 differently regulates the transcriptional levels of type I collagen and stromelysin-1 gene and denoting IL-10 seems to take part in the homeostasis of ECM at pretranscriptional level.

Effects of the cyclic-tension force on the expression of MMP-3 mRNA in rat bone marrow osteoclasts / 实用口腔医学杂志

Objective:To study the effect of the cyclic-tension force on the expression of matrix metalloproteinase-3 ( MMP-3) mRNA in osteoclasts. Methods: Bone marrow cells were cultured and induced by the combination of IL-6 and 1,25(OH) 2D 3 and identified. The bone marrow cells were seeded onto elastic 6-well culture plate at a density of 2?105 cells/ml in 2ml in each well and cultured for 7 days. Then the cells were subjected to 12% elongation by a strain unit at 6 cycles/min (i.e.5-s elongation and 5-s relaxation) . After 24 hours of stretching, the expression of MMP-3 mRNA in osteoclasts were determined by in situ hybridization analysis. Results:The stretched osteoclasts showed enhanced MMP-3 mRNA expression level. Conclusion: The mechanical stretching may affect the bone-resorbing activity by up-regulated the expression of MMP-3 mRNA in osteoclasts.