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1.
Article in Chinese | WPRIM | ID: wpr-905900

ABSTRACT

Objective:To illustrate the effect of M1/M2 polarization of macrophages on gouty arthritis models induced with monosodium urate and reveal the molecular mechanism of total saponins from Dioscoreae Nipponicae Rhizoma to treat gouty arthritis. Method:A total of 72 male SD rats were randomly divided into four groups: normal group, model group, total saponin group (160 mg·kg<sup>-1</sup>), celecoxib group (43.3 mg·kg<sup>-1</sup>), with 18 rats in each group. Gouty arthritis models were induced by injecting monosodium urate into ankle joints bilaterally. Histopathology changes of ankle joints were observed by hematoxylin-eosin(HE) staining. Immunohistochemistry method was used to detect the protein expression change of CD68, interleukin-4(IL-4), inducible nitric oxide synthase (iNOS) and transforming growth factor-<italic>β</italic><sub>1</sub>(TGF-<italic>β</italic><sub>1</sub>). Result:HE staining results showed that the inflammation of the model group was most obvious on the third day after modeling, and the disease was in the acute stage. On day 5, the inflammation was alleviated, and on day 8, the inflammation was still present but close to normal. The total saponin group and celecoxib group could improve the pathological changes of synovial tissue, and the effect of total saponin group was more obvious. Immunohistochemical results were as follows. Compared with the normal group. The expression of CD68 and iNOS in the model group increased on the 3rd,5th and 8th day of administration (<italic>P</italic><0.01). Compared with the model group, the total saponins group could reduce the expression of CD68 and iNOS (<italic>P</italic><0.05,<italic>P</italic><0.01)on the 3rd day of administration, and significantly reduced them expression on the 5th and 8th days (<italic>P</italic><0.01). Compared with the normal group, IL-4 and TGF-<italic>β</italic><sub>1</sub> expression were increased in the model group when the drug was given for three days(<italic>P</italic><0.01). Total saponin group could enhance IL-4 expression(<italic>P</italic><0.05)and decreased the TGF-<italic>β</italic><sub>1</sub> expression(<italic>P</italic><0.01). Compared with normal group, the expression of IL-4 in the model group decreased on the 5th and 8th day of administration (<italic>P</italic><0.01), and the expression of TGF-<italic>β</italic><sub>1</sub> in the model group decreased on the 5th day of administration(<italic>P</italic><0.01). Compared with the model group, the total saponins group could increase the expression of IL-4 and TGF-<italic>β</italic><sub>1</sub> at 5 d and 8 d after administration (<italic>P</italic><0.01). Conclusion:Total saponins from Dioscoreae Nipponicae Rhizoma has the potential effect to treat gouty arthritis by regulating M1/M2 polarization.

2.
Organ Transplantation ; (6): 421-2021.
Article in Chinese | WPRIM | ID: wpr-881526

ABSTRACT

Objective To evaluate the effect of interleukin (IL)-10 on donor lung function after ex vivo lung perfusion (EVLP) in rats of cardiac death. Methods Twenty adult male SD rats were randomly divided into the simple perfusion group (group A, n=10) and modified perfusion group (group B, n=10). Perfusate A (without IL-10) and perfusate B (supplemented with IL-10) was administered in group A and B, respectively. The EVLP rat models of cardiac death were established. The appearance of donor lung, dry-to-wet (D/W) ratio of donor lung tissues, the function and metabolism of donor lung, the morphology of donor lung and the levels of inflammatory markers of donor lung were statistically compared between two groups. Results After perfusion, evident edema of the whole donor lung, poor compliance and a large amount of edema fluid discharged from the airway were observed in group A, whereas no obvious edema and good compliance were found in group B. Compared with group A, the D/W ratio of lung tissues in group B was higher (P < 0.05). In both groups, the pulmonary vein partial pressure of oxygen reached the peak at 2 h after perfusion, which did not significantly differ between two groups (P > 0.05). In group B, the pulmonary artery pressure was increased at a lower speed and significantly lower after perfusion, and the lactic acid level in the perfusate was significantly lower than those in group A (all P < 0.05). In group A, the alveolar structure was largely destroyed and the cells was rare. In group B, the alveolar structure was relatively normal without evident cell edema. The incidence of cell apoptosis of donor lung was high in group A, whereas no obvious cell apoptosis of donor lung was noted in group B. After perfusion for 4 h, the levels of monocyte chemoattractant protein (MCP)-1 and IL-6 were significantly increased, the IL-4 levels were remarkably decreased (all P < 0.05), but the levels of tumor necrosis factor (TNF)-α, IL-1α and inducible nitric oxide synthase (iNOS) did not significantly change in both groups (all P > 0.05). Conclusions IL-10 may improve the function of donor lung after EVLP in rat of cardiac death by reducing cell apoptosis.

3.
Article in Chinese | WPRIM | ID: wpr-872950

ABSTRACT

Objective::To investigate the effect of betulic acid(BA) on steatosis LO2 cells. Method::LO2 cells were intervened with BA at different gradient concentrations (0, 10, 20, 40, 80, 160, 250 μmol·L-1) for 24 hours. methyl thiazolyl tetrazolium(MTT) staining was used to observed cell viability to determine the final concentration of BA. The cells were divided into control, model, dimethylsulfoxide (DMSO) and BA groups, as well as BA groups intervened with low, middle and high concentrations. First, model, DMSO and BA group's cells were cultured in 10% Lipid Mix 1 medium for 24 hours to establish a nonalcoholic fatty liver model. Then, DMSO group and low, medium and high-concentration groups were separately cultured with 0.1%DMSO medium and 20, 40, 80 μmol·L-1 BA medium for 24 hours. And control and model groups were cultured in drug-free medium for 24 hours. Oil red O staining and Nile red staining were used to observe the intracellular lipid droplets. Immunofluorescence was used to detect the protein expression of inducible nitric oxide synthase (iNOS). Western blot was used to detect the protein expression levels of receptor for advanced glycation end-products (RAGE), nuclear factor κB p65 (NF-κB p55) and iNOS. Result::BA within the concentration of 80 μmol·L-1 had no significant toxicity on LO2 cells. Compared with control group, the intracellular lipid droplets were significantly increased in the model group, and the expressions of oxidative stress-related proteins RAGE, NF-κB p65 and iNOS also increased significantly(P<0.05). Compared with model group, the intracellular lipid droplets in DMSO group were similar to those in model group, with no significant difference in the three protein expressions between the two groups. However, the intracellular lipid droplets deposition in the BA group was significantly decreased. And the expressions of RAGE, NF-κB p65 and iNOS proteins in high-concentration BA group were significantly decreased(P<0.05, P<0.01). Conclusion::BA can significantly improve the intracellular fat deposition in LO2 cells, which was probably related to the inhibition of the expressions of oxidative stress-related proteins RAGE, NF-κB p65 and iNOS.

4.
Article in Chinese | WPRIM | ID: wpr-802419

ABSTRACT

Objective: To evaluate the effect of Houpu Sanwu Tang on the postoperative ileus (POI), and observe its underlying mechanisms of action on interstitial cells of cajal (ICC) and inducible nitric oxide synthase(iNOS) regulation of POI. Method: Totally 87 healthy adult male SD rats were randomly divided into sham operation group, saline control group and Houpu Sanwu Tang group at low, medium and high doses. Houpu Sanwu Tang low, middle and high dose groups received orally Houpu Sanwu Tang(2.25,4.5,9 g·kg-1); Sham operation group (Sham operation) and saline control group (Saline control) received orally normal saline. Surgical procedure was used to induce the postoperative ileus. Changes in intestinal propulsion rate, intestinal mucosal injury and small intestine expression of c-kit and iNOS among these groups were detected. Result: Intestinal propulsion rate was significantly higher in Houpo Sanwu Tang group than that in Saline control group (PPPPConclusion: Houpu Sanwu Tang can improve the intestinal propulsion rate and the recovery in POI rats. The mechanisms are related to the inhibition of the generation of iNOS, the alleviation of inflammatory response, and increase of the number of ICC, so as to ensure its normal function, and improve the intestinal dynamic disorder.

5.
Clinical Medicine of China ; (12): 249-252, 2018.
Article in Chinese | WPRIM | ID: wpr-706662

ABSTRACT

Objective To observe the effect of HIF-1a/iNOS signaling pathway on myocardial ischemia-reperfusion injury in rat heart transplantation and the protective mechanism of N-acetylcysteine (NAC) on donor heart after cardiac transplantation in rats.Methods Eighty healthy male Lewis rats were randomly divided into 3 groups,the control group (0.3 ml saline was infused via inferior vena cava 30 min before donor harvest or implantation),NAC donor pretreatment group [NAC (30 mg/kg.w) was injected into the vena cava of donor rat 30 min defore donor harvest],and the NAC receptor pretreatment group (NAC 300 mg/kg.w was injected into the vena cava of the recipient rats 30 min before transplantation.The 30 min was injected into the vena cava of the recipient rats).A transplant model was established and the graft was obtained after 24 h transplantation.The expression of iNOS,HIF-1a and mRNA in cardiac muscle tissue was detected by immunohistochemistry and Real time-PCR.Results HIF-1a protein expression in graft myocardial tissue was significantly lower in NAC donor pretreatment and recipient pretreatment group compared with control group (P <0.05),the differences were statistically significant (2.72±0.17 vs.2.24±0.23 vs.3.14.±0.16,F=56.26,P =0.000).The iNOS protein expression in NAC donor pretreatment group,and NAC recipient pretreatment group were lower than that in the control group (1.52±0.18 vs.1.61±0.19 vs.3.30±0.18,F=232.345,P =0.000),the differences were statistically significant (P < 0.05).24 h after transplantation,the differences in graft myocardial tissue HIF-1a and iNOS mRNA among the three groups were statistically significant (F=7.467,16.490,P=0.003,0.000).The expression of iNOS mRNA in the NAC receptor pretreatment group was significantly lower than that in the control group (P<0.05).Conclusion HIF-1a/iNOS signaling pathway can regulate ischemia reperfusion injury in rat heart transplantation,and the protective effect of NAC on donor heart maybe mediated via this pathway.

6.
Acupuncture Research ; (6): 226-230, 2018.
Article in Chinese | WPRIM | ID: wpr-844467

ABSTRACT

OBJECTIVE: To observe the effect of acupuncture intervention on depressive-like behavior and nuclear factor-κB (NF-κB), inducible nitric oxide synthase (iNOS) and nitric oxide (NO) levels in the prefrontal cortex in chronic stress induced depression rats, so as to explore its mechanism underlying antidepressant effect. METHODS: SD rats were randomly divided into control, model, acupuncture and medication (Fluoxetine) groups (8 rats/group). The depression model was established by chro-nic unpredictable stress stimulation (fasting, water deprivation, cold water swimming, tail clamping, constraining, etc., and solitary raising) for 28 days. Acupuncture treatment was applied to "Baihui" (GV 20) and unilateral "Neiguan"(PC 6) once every other day for 14 days. Fluoxetine (10 mg/kg) was given to rats of the medication group by gavage (p.o.) before every stress stimulation, once every day for 28 days. The animals' behavior was tested by using sucrose intake and open field tests (number of crossings and rearings in 5 min). The expression of NF-κB in the prefrontal cortex was detected by Western blot, and the contents of prefrontal iNOS and NO were detected by ELISA and nitric reductases, respectively. RESULTS: After modeling, the sucrose intake and the numbers of crossing and rearing times were significantly decreased (P0.05). CONCLUSION: Acupuncture treatment can improve the depressive-like behavior of depression rats, which is closely related to its effects in down-regulating the levels of NF-κB protein, iNOS and NO in the prefrontal cortex to reduce brain inflammatory damage.

7.
Braz. j. pharm. sci ; 49(spe): 57-65, 2013. ilus
Article in English | LILACS | ID: lil-686586

ABSTRACT

Dendrimers constitute an intriguing class of macromolecules which find applications in a variety of areas including biology. These hyperbranched macromolecules with tailored backbone and surface groups have been extensively investigated as nanocarriers for gene and drug delivery, by molecular encapsulation or covalent conjugation. Dendrimers have provided an excellent platform to develop multivalent and multifunctional nanoconjugates incorporating a variety of functional groups including drugs which are known to be anti-inflammatory agents. Recently, dendrimers have been shown to possess anti-inflammatory properties themselves. This unexpected and intriguing discovery has provided an additional impetus in designing novel active pharmaceutical agents. In this review, we highlight some of the recent developments in the field of dendrimers as nanoscale anti-inflammatory agents.


Dendrímeros constituem uma classe intrigante de macromoléculas que apresentam aplicações em diversas áreas incluindo biologia. Essas macromoléculas extremamente ramificadas com esqueleto planejado e grupos de superfície foram extensivamente investigadas como nanotransportadores de genes e de fármacos, por encapsulamento molecular ou conjugação covalente. Dendrímeros têm proporcionado uma plataforma excelente de desenvolvimento nanoconjugados multivalentes e multifuncionais incorporando uma variedade de grupos funcionais, incluindo fármacos que são conhecidos por atuarem agentes antiinflamatórios. Recentemente, os dendrímeros mostraram propriedades antiinflamatórias. Esta inesperada e intrigante descoberta tem proporcionado um impulso adicional no planejamento de novos agente farmacêuticos ativos. Nesta revisão, nós destacamos alguns dos desenvolvimentos recentes no campo dos dendrímeros como agentes antiinflamatórios em nanoescala.


Subject(s)
Anti-Inflammatory Agents/analysis , Dendrimers/analysis , Cytokines , Nitric Oxide Synthase/metabolism
8.
Article in English | WPRIM | ID: wpr-116724

ABSTRACT

Inducible nitric oxide synthase (iNOS) is a main enzyme producing nitric oxide during inflammation and thus contributes to the initiation and development of inflammatory cardiovascular diseases such as atherosclerosis. Epigallocatechin-3-gallate (EGCG), the major catechin derived from green tea, has multiple beneficial effects for treating cardiovascular disease, but the effect of EGCG on the expression of vascular iNOS remains unknown. In this study, we investigated (i) whether EGCG inhibits the expression of vascular iNOS induced by angiotensin II in human umbilical vein endothelial cells and, if it does inhibit, (ii) mechanisms underlying the inhibition. Angiotensin II increased expression levels of vascular iNOS; EGCG counteracted this effect. EGCG increased the production of reactive oxygen species. Moreover, EGCG did not affect the production of reactive oxygen species induced by angiotensin II. These data suggest a novel mechanism whereby EGCG provides direct vascular benefits for treating inflammatory cardiovascular diseases.


Subject(s)
Angiotensin II , Atherosclerosis , Cardiovascular Diseases , Catechin , Human Umbilical Vein Endothelial Cells , Humans , Inflammation , Nitric Oxide , Nitric Oxide Synthase Type II , Reactive Oxygen Species , Tea
9.
Article in Korean | WPRIM | ID: wpr-109513

ABSTRACT

INTRODUCTION: Heat shock protein70 (HSP70) is a highly conserved family of proteins produced after a variety of stresses. Many studies reported that the overexpression of HSP70 can improve the prognosis of the patients with sepsis through a reduction of the nitric oxide concentration. However, these results only revealed the effect of HSP70 and nitric oxide. No studies have examined the relationship between HSP70 and nitric oxide. The aim of this study was to evaluate the effect of the overexpression of HSP70 on the expression of inducible nitric oxide synthase and the nitric oxide concentration. In addition, the mechanism of the relationship of HSP70 and inducible nitric oxide synthase (iNOS) in sepsis was examined. MATERIALS AND METHODS: The experiments were performed on male sprague-dawley rats. Sepsis was induced by a cecal ligation and puncture (CLP). Glutamine (GLN) or saline was administered 1 hour after the initiation of sepsis. Serum and lung tissues were acquired from the rats 12 hours or 24 hours after the initiation of sepsis. The nitric oxide concentration, the expression of HSP70 in lung, and the gene expression of iNOS in lung were analyzed. The three groups, sham operation, CLP and CLP+GLN, were compared. RESULTS: Compared to the other groups, in CLP+GLN, GLN administered after the initiation of sepsis enhanced the expression of HSP70 in the lung at 12 hours (47.19+/-10.04 vs. 33.22+/-8.28, P=0.025) and 24 hours (47.06+/-10.60 vs. 31.90+/-4.83, P=0.004). In CLP+GLN, GLN attenuated the expression of iNOS messenger RNA (mRNA) in the lung at 12 hours (5,513.73+/-1,051.60 vs. 4,167.17+/-951.59, P=0.025) and 24 hours (18,740.27 +/-8,241.20 vs. 9,437.65+/-2,521.07, P=0.016), and reduced the concentration of nitric oxide in the serum at 12 hours (0.86+/-0.48 vs. 3.82+/-2.53, P=0.016) and 24 hours (0.39+/-0.25 vs. 1.85+/-1.70, P=0.025). CONCLUSION: The overexpression of HSP70 induced by the administration of GLN in sepsis attenuates the expression of the iNOS gene but reduces the nitric oxide concentration.


Subject(s)
Animals , Gene Expression , Glutamine , Heat-Shock Proteins , Hot Temperature , HSP70 Heat-Shock Proteins , Humans , Ligation , Lung , Male , Nitric Oxide , Nitric Oxide Synthase , Nitric Oxide Synthase Type II , Prognosis , Proteins , Punctures , Rats , Rats, Sprague-Dawley , RNA, Messenger , Salicylamides , Sepsis , Shock
10.
Article in Chinese | WPRIM | ID: wpr-405822

ABSTRACT

dosage ( P<0.01).Conclusion UVA can inhibit the proliferation activity of human skin fibroblasts. It might be related to the up-regulation of iNOS gene expression and the over-secretion of NO induced by UVA.

11.
China Oncology ; (12): 899-903, 2009.
Article in Chinese | WPRIM | ID: wpr-404771

ABSTRACT

Background and purpose: There have been no reports on the clinical and biological significance of the co-expression of inducible nitric oxide synthase (iNOS) and β-catenin (β-cat) in has.pharyngeal carcinoma(NPC). This study was aimed to investigate the expression of iNOS and β-cat in NPC, to analyze their interrelation, and to explore their roles in the carcinogenesis, development, invasion, and metastasis of NPC. Methods: The expression of iNOS and β-cat was examined in 50 poorly differentiated NPC and 15 normal nasopharyngeal epithelium tissue by immunohistochemical staining (SP method). None of the patients had received radiotherapy and chemotherapy. The results of immunostaining were observed by two pathologists independently, using double blind scoring method. iNOS and β-cat expression were categorized by the extent and intensity of staining using a semiquantitative method. Results: No iNOS expression was observed in 15 normal nasopharyngeal epithelium, but β-cat expression was located in cytomembrane in normal samples. The positive rate of iNOS protein expression was 74.0% (37/50) in NPC tissues. The expression of iNOS was statistically different among the NPC group and normal nasopharyngeal epithelium group, T_(1-2) group and T_(3-4) group, metastatic lymph nodes group and no metastatic lymph nodes group. There was a great quantity of β-cat expression in cytoplasm, little or no β-cat expression in cytomembrane and nucleus in NPC tumorous tissues. Statistical analysis indicated that strong positive expression of β-cat in cytoplasm was significantly higher in NPC than those of normal nasopharyngeal epithelium. The positive expression of β-cat was significantly correlated with clinical stage, invasion and lymph gland metastasis. Neither iNOS nor β-cat was significantly different between different ages and genders. There was a positive relationship between iNOS and β-cat expression in NPC tissues (r=0.394, P=0.005). Conclusion: The positive expressions of iNOS and β-cat in cytoplasm were significantly correlated with clinical stage, invasion and metastasis, and were not significantly different in terms of ages and genders of the patients. There was a positive relationship between the expression of iNOS and β-cat in NPC tissues. Co-overexpression of iNOS and β-cat may play an important role in carcinogenesis, development, invasion, and metastasis of NPC.

12.
Article in Korean | WPRIM | ID: wpr-66136

ABSTRACT

BACKGROUND: Near-infrared light (NIR, 0.8-1.5 micrometer light) has been used in therapeutic devices for various injuries such as infected, ischemic and hypoxic wound. NIR-emitting technology has been developed recently in Korea. We hypothesized that NIR may have an anti-inflammatory effect and investigated the effect of NIR-irradiated media on cell culture. METHODS: Three kinds of cell lines, CAPE (vascular endothelial cell), NIH3T3 (fibroblast), and RD (smooth muscle cell) cells were cultured for 4 days in 10% FBS-containing media (1x10(4) cells/well), which were irradiated or not irradiated (control) by Eco-NFIR Drive (Model #0210, Ecowavetech, Korea). The cells were stimulated by 10 mcg/mL of bacterial lipopolysaccharide (LPS) or sodium nitroprusside (SNP). Cellular proliferation was measured by methylthiazol tetrazolium assay. Expression of interleukin (IL)-1 beta and nitric oxide was measured by ELISA. Expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) was measured by immunofluorescence staining. RESULTS: NIR-irradiated medium was favorable for CAPE cell proliferation (N=8, P=0.000). IL-1 beta secretion from LPS-stimulated NIH3T3 cells incubated in the NIR medium was below that of control medium (N=4, P=0.026). Nitrate production seemed to be low in NIR-irradiated medium although statistically insignificant (N=4, P=0.076). Expression of iNOS of the LPS-stimulated cells was decreased in NIR medium, however, Cox-2 expression was not different between the two media. CONCLUSIONS: NIR-irradiated medium supported vascular endothelial cell proliferation and showed an anti-inflammatory effect on fibroblast culture. These results can be used as basic data for future research on the clinical application of NIR.


Subject(s)
Animals , Anti-Inflammatory Agents/chemistry , Cattle , Cell Line , Culture Media , Cyclooxygenase 2/metabolism , Humans , Infrared Rays , Interleukin-1beta/metabolism , Lipopolysaccharides/pharmacology , Mice , Nitric Oxide/metabolism , Nitric Oxide Synthase Type II/metabolism
13.
China Oncology ; (12)2006.
Article in Chinese | WPRIM | ID: wpr-545391

ABSTRACT

Background and purpose:Concurrent chemo-radiotherapy(CCRT) was considered the best treatment plan for advanced nasopharyngeal carcinoma(NPC),but there was no uniform conclusion as to which category of patients and which chemotherapy associated radiotherapy would have the best therapeutic effect. As the standard treatment plan for advanced NPC, DDP concurrent chemo-radiotherapy was recommended by some scholars. DDP can raise the expression of inducible nitric oxide synthase(iNOS) protein and synthesize nitric oxide (NO) with anti-tumor effects, so we considered whether the therapeutic effect could be predicted and the corresponding treatment plan could be selectived to detect the iNOS expression in the pretherapy NPC tissues.The purpose of this study was to investigate the relation between the expression of iNOS protein and the nasopharyngeal tumor with complete response or with residue after DDP concurrent chemo-radiotherapy, so that the most appropriate plan of treatment can be adopted and the complete response rate of nasopharyngeal tumor can be raised. Methods:All patients were poorly differentiated NPC.The expression of iNOS protein was examined in 30 patients of nasopharyngeal tumor with complete response and 30 patients with residual tumor after DDP concurrent chemo-radiotherapy by immunohistochemical staining (SP method).None of the patients had received radiotherapy and chemotherapy.Results:Immunohistochemical examination revealed that iNOS expression in the NPC tissues was located in both the nucleus and cytoplasm of the tumorous tissues. The intensity of iNOS expression was stronger in the nucleus than in the cytoplasm of the tumorous tissues.The positive rates of iNOS protein expressions were 71.67%(43/60) in NPC tissues. It was 86.67% and 53.33% in 30 tumors with complete response and with residual tumor, respectively. The difference was statistically significant.The rate of iNOS strong postive expressions in the group of residual tumors was higher than that of the group with complete response. It was statistically different,but weak and moderate postive expressions did not have statistical difference.Conclusions:According to the difference of iNOS expression, it is a valuable method to select the most appropriate plan of treatment and the complete response rate of nasopharyngeal tumor can be raised.

14.
Article in Korean | WPRIM | ID: wpr-51455

ABSTRACT

BACKGROUND: Particulate matters (PM) when inhaled is known to induce pulmonary diseases including asthma and chronic bronchitis when inhaled. Despite the epidemiological proofevidence, the pathogenesis of PM-related pulmonary diseases is unclearremain poorly understood. METHODS: Primary alveolar macrophages were harvested from the SPF and inflammatory rats by bronchioalveolar lavage (BAL). The cultured primary alveolar macrophages were treated with the medium only, PM only (5~40 microgram/cm2), LPS (5ng/ml) only, and PM with LPS for 24 and 48 hours. The level of secreted nitric oxide (NO) was assayed from the cultured medium by using the Griess reaction. The cultured cells were utilized for the western blotting against the inducible nitric oxide synthase (iNOS) proteins. Immunocyto- chemical staining against the iNOS and NT-proteins were performed in cells that cultured in the Lab-Tek(R) chamber slide after treatments. RESULTS: The PM that utilizein this experiments induced NO formation with iNOS expression in the cultured SPF and inflammatory rats alveolar macrophages, by itself. When the cells were co-treated with PM and LPS, there was a statistically significant synergistic effect on NO formation and iNOS expression over the LPS effect. The cells from the sham control showed minimal immunoreactivity for the NT-proteins. Significantly higher quantities of NT-proteins were detected in the PM and PM with LPS co-treated cells than from the sham control. CONCLUSION: Increased iNOS expression and NO formation with increased NT-proteins formation might be involved in the pathogenesis of PM-induced lung injury.


Subject(s)
Animals , Asthma , Blotting, Western , Bronchitis, Chronic , Bronchoalveolar Lavage , Cells, Cultured , Lung Diseases , Lung Injury , Macrophages, Alveolar , Nitric Oxide Synthase Type II , Nitric Oxide , Rats
15.
China Pharmacy ; (12)2005.
Article in Chinese | WPRIM | ID: wpr-530836

ABSTRACT

OBJECTIVE:To observe the inhibitory effect of phenolicalkaloids of menispermum dauricum(PAMD)on the human pancreatic cell line(BXPC-3)tumor and study the effect of PAMD on serum content of inducible nitric oxide synthase(iNOS)in tumor-bearing mice.METHODS:The BXPC-3 tumor-bearing mice model was established,and the model mice were randomly assigned to 6 groups:model control group,blank control group,cyclophosphamide group,3 PAMD groups(high,medium and low dosages).The blank control group was not inoculated with tumor strain but given same volume of normal saline.The tumor-inhibition ratio of PAMD was detected and the content of iNOS of in peripheral blood of tumor-bearing mice was determined by fluorospectrophotometry.RESULTS:PAMD(high,medium and low dosages)showed marked inhibitory effect on tumor growth of BXPC-3 cell line in tumor-bearing mice,with the inhibition ratios at 34.91%,52.83% and 41.51%,respectively.As compared with model control group,PAMD-treated groups showed significantly lower content of iNOS in peripheral blood of tumor-bearing mice.CONCLUSION:PAMD showed marked inhibitory effect on tumor growth of BXPC-3 cell line in tumor-bearing mice,which might be attributed to the lowered content of iNOS.

16.
Article in Korean | WPRIM | ID: wpr-76221

ABSTRACT

PURPOSE: Intimal hyperplasia is a pathologic condition that is commonly observed with the atherosclerotic change of blood vessels; this condition is usually observed after arterial operation and interventions such as angioplasty, endaterectomy and bypass grafting, and it remains a major cause for hindering for arterial patency. It is a multiple processes that includes smooth muscle cell migration, proliferation, and expression of extracellular matrix proteins, such as fibronectin. Nitric oxide (NO) has been found to possess many different vasoprotective properties: inhibition of platelet aggregation and adherence, inhibition of leukocyte chemotaxis, inhibition of vascular smooth muscle cell proliferation and migration, inhibition of endothelial cell apoptosis and stimulation of endothelial cell growth. Overexpression of nitric oxide synthase (NOS) in the vascular wall has been used to regulate vasomotor function, prevent neointimal formation after balloon injury or vein grafting, and to prevent transplant vasculopathy. NOS gene transfer to the vascular wall holds great promise as a means of controlling local vascular function. METHOD: We investigated whether the inducible NOS (iNOS) gene transfer to the arterial wall has an inhibitory effect on intimal hyperplasia after endothelial denudation and on the change of the level of extracellular matrix fibronectin expression in the rabbit common carotid artery. Rabbits were divided into three groups: the saline only (without injury) normal group, the injury + saline intima injury group, and the injury + recombinant adenoviral vector encording human iNOS (AdiNOS) gene transfer group (n=5 per group). AdiNOS (1.6x10(10) plaque-forming units [pfu]) were used for the iNOS gene delivery and the virus was intraluminally infected to the balloon-injured common carotid arterial wall for 20 minutes. The NO levels were assessed in the blood of all the animals at the three time point: before-, 1 hr after-, and 14 days after the surgical procedures. The left common carotid arteries were harvested from the animals at 14 days after balloon injury and they were then assessed for fibronectin expression and intimal hyperplasia. RESULT: 14 days after AdiNOS infection, the NO levels were increased 67% in the gene transfer group compared with those levels obtained before the gene transfer (333.5+/-11.5 vs. 557.4+/-72.1, respectively, P<0.05), and the intima/intima+media ratio and the FN expression were reduced by 60% and 50%, respectively, in the gene transfer group compared with the intima injury group (41.5+/-5.6 vs. 16.6+/-2.8, respectively, P<0.05 and 307.9+/-38.3 vs. 154.3+/-20.2, respectively, P<0.05). CONCLUSION: Our results show that iNOS gene transfer to the arterial wall resulted in increased NO levels in the blood, and the procedure greatly inhibited intimal hyperplasia and it reduced fibronectin expression in the arteriaql wall.


Subject(s)
Angioplasty , Animals , Apoptosis , Blood Vessels , Carotid Arteries , Carotid Artery, Common , Cell Proliferation , Chemotaxis, Leukocyte , Endothelial Cells , Extracellular Matrix , Extracellular Matrix Proteins , Fibronectins , Humans , Hyperplasia , Muscle, Smooth, Vascular , Myocytes, Smooth Muscle , Nitric Oxide , Nitric Oxide Synthase , Nitric Oxide Synthase Type II , Platelet Aggregation , Rabbits , Transplants , Veins
17.
Article in Korean | WPRIM | ID: wpr-34545

ABSTRACT

BACKGROUND: Nitric oxide(NO) produced by activated macrophages through the action of iNOS is the key molecule in the killing mycobacterium. Prostaglandins produced by the action of COX-2, also, are the important mediators of inflammation and other pathophysiologic process. A complex relationship is emerging with regard to "cross-talk" between the NO and COX-2 pathways. OBJECTIVE: The purposes of this study were to investigate the expression of iNOS and COX-2 across the spectrum of leprosy in the paraffin-embedded skin lesions, to demonstrate the interaction between iNOS and COX-2 expression, and to demonstrate the differences in the cell types expressing the iNOS or COX-2. METHOD: In the paraffin-embedded skin lesions of 30 new cases of leprosy(TT, n=4; BT, n=4; BL, n=7; LL, n=15), iNOS and COX-2 expression were detected by using immunohistochemical staining. RESULTS: iNOS expression was 2.0-55.8%(mean 15.9%) and the level of expression of iNOS in TT(31.2%) and BT(32.6%) lesions was significantly higher than that of BL(11.1%) and LL(8.6%) lesions(p<0.05). COX-2 expression was 3.6-74.5%(mean 27.1%) and the level of expression of COX-2 in TT(59.2%) lesions was significantly higher than that of BT, BL and LL lesions(p<0.05). There was positive correlation between iNOS and COX-2 expression, that is, the lesions which expressed high level of iNOS also expressed COX-2 highly. The correlation was statistically significant(r=0.535, p<0.05). The overall level of COX-2 expression(27.1%) was higher than that of iNOS expression(15.9%), and when compared the expression of them across the spectrum of leprosy, COX-2 expressed higher than iNOS in TT and LL lesions. CONCLUSION: Both iNOS and COX-2 were expressed in all types of leprosy skin lesions and the level of iNOS expression in TT and BT lesions was significantly higher than that of BL and LL lesions. The level of expression of COX-2 in TT lesions was significantly higher than that of BT, BL and LL lesions. These results suggest that iNOS and COX-2 have important roles in anti-mycobacterial activities in leprosy lesions. The positive correlation between iNOS and COX-2 expression suggests that NO and COX-2 might interact synergistically or additively rather than suppress each other.


Subject(s)
Homicide , Humans , Inflammation Mediators , Leprosy , Macrophages , Mycobacterium , Prostaglandins , Skin
18.
Article in Korean | WPRIM | ID: wpr-78823

ABSTRACT

BACKGROUND: Nitric oxide is a short-lived effector molecule derived from L-arginine by the nitric oxide synthase(NOS). Nitric oxide plays a role in a number of physiologic and pathophysiologic functions including host defense, edema formation, and regulation of smooth muscle tone. Some kinds of cells including macrophage are known to produce large quantities of nitric oxide in response to inflammatory stimuli such as interleukin-1beta(IL-1beta), tumor necrosis factor-alpha(TNF-alpha), interferon-gamma(IFN-gamma) and lipopolysaccharide(LPS). Reactive oxygen species are also known to be important in the pathogenesis of acute cell and tissue injury such as acute lung injury model. METHODS: Using the RAW264.7 cells, we have examined the ability of oxidant hydrogen peroxide(H2O2) to stimulate nitric oxide production and inducible NOS mRNA expression. Also, we have examined the effects of NOS inhibitors and antioxidants on H2O2 induced nitric oxide production. RESULTS: Stimulation of RAW264.7 cells with combinations of 100 ng/ml IL-1beta, 100 ng/ml TNF-alpha, and 100 U/ml IFN-gamma or 100 U/ml IFN-gamma and 1 microg/ml LPS induced the synthesis of nitric oxide as measured by the oxidation products nitrite(NO2-) and nitrate(NO3-). Addition of 250 microM- 2 mM H2O2 to the cytokines significantly augmented the synthesis of NO2- and NO3-(p<0.05). When cells were incubated with increasing concentrations of H2O2 in the presence of IL-1beta, TNF-alpha and IFN-gamma at constant level, the synthesis of NO2- and NO3- was dose-dependently increased(p<0.05). 3. NG-nitro-L-arginine methyl ester(L-NAME), dose dependently, significantly inhibited the formation of NO2- and NO3- in cells stimulated with LPS, IFN-gamma and H2O2 at constant level(p<0.05). 4. Catalase significantly inhibited the H2 O2-induced augmentation of cytokine- induced NO2- and NO3- formation(p<0.05). But, boiled catalase did not produce a significant inhibition in comparison with the native enzyme. Another antioxidant 2-mercaptoethanol and orthophenanthroline dose-dependently suppressed NO2- and NO3- synthesis(p<0.05). Northern blotting demonstrated that H2O2 synergistically stimulated the cytokine-induced iNOS mRNA expression in RAW264.7. CONCLUSION: These results suggest that H2O2 contributes to inflammatory process by augmenting the iNOS expression and nitric oxide synthesis induced by cytokines.


Subject(s)
Acute Lung Injury , Antioxidants , Arginine , Blotting, Northern , Catalase , Cytokines , Edema , Hydrogen Peroxide , Hydrogen , Macrophages , Mercaptoethanol , Muscle, Smooth , Necrosis , Nitric Oxide , Nitric Oxide Synthase Type II , Nitroarginine , Reactive Oxygen Species , RNA, Messenger , Tumor Necrosis Factor-alpha
20.
Article in Chinese | WPRIM | ID: wpr-670914

ABSTRACT

Objective:To study the changes of iNOS expression in periodontal tissues of rats with periodontitis in orthodontic tooth movement. Methods:48 SD rats at 10 weeks were divided randomly into two groups: periodontitis teeth group and normal teeth group. The maxillary first molar was pulled mesially both in the two groups. 4 rats were executed each time after activation of pull-spring 1, 2, 3, 7,14 and 21d both in two groups. iNOS immunohistochemistry staining were done to compare the expression variations. Results: There were significant differences between the two groups in 2,3, 7, 14 and 21 d. The periodontitis teeth group had stronger positive signals. Conclusion: Periodontitis will increase the expression of iNOS in the rats periodontal tissue during tooth movement, and influence the reconstruction of periodontium.

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