1.Airway neutrophils apoptosis in children with severe asthma.
Jing LIAO ; Guang-min NONG ; Min JIANG ; Yu-xian ZHANG ; Xiu-an LIANG ; Jing LIU
Chinese Journal of Pediatrics 2009;47(8):628-631
OBJECTIVETo investigate the changes of neutrophils in airway inflammation in children with severe asthma.
METHODChildren with mild to moderate asthma (n=23), severe asthma (n=16) and healthy control subjects (n=16) underwent lung function tests and sputum induction. The sputum specimens were assayed for cellular differential count, the supernatant and peripheral blood were assayed for the concentrations of IL-8 by "sandwich" enzyme linked immunosorbent assay (ELISA). Sputum supernatant, IL-8 and mifepristone were assessed for their abilities to prolong the in vitro survival of blood-derived neutrophils.
RESULTThe percentage of sputum neutrophils was significantly higher in severe asthmatics [59.54 (41.99-74.65)%] than mild-moderate asthmatics [30.03 (15.94-47.71)%] and healthy control subjects [29.72(16.53-45.74)%] (P < 0. 01); the level of IL-8 in sputum was significantly higher in severe asthmatics [2907.78 (331.67 - 3457.93) ng/L] than mild-moderate asthmatics [287.58 (130.75-656.84) ng/L] and healthy control subjects [179.2 (58.55-346.59) ng/L] (P < 0.01); the percentages of neutrophilic apoptosis respectively cultured with LPS [(10.57 +/- 1.97)%], severe asthmatics supernatant [(11.82 +/- 2.96)%], IL-8 [(10.47 +/- 1.93)%], dexamethasone [(9.93 +/- 1.95)%], severe asthma supernatant + mifepristone [(12.15 +/- 2.86)%] in vitro were lower than that cultured with PBS [(17.98 +/- 2.27)%], healthy control supernatant [(17.37 +/- 2.50)%], mild-moderate asthmatics supernatant [(16.35 +/- 3.26)%], mifepristone [(17.89 +/- 2.38)%], and dexamethasone + mifepristone [(17.06 +/- 2.59)%] (P < 0.01).
CONCLUSIONSuppression of neutrophilic apoptosis seems to play a potential role in airway neutrophilic inflammation in severe asthmatics, and the level of IL-8 in sputum was significantly higher in patients with severe asthmatics.
Adolescent ; Apoptosis ; Asthma ; metabolism ; pathology ; physiopathology ; Case-Control Studies ; Child ; Female ; Humans ; Inflammation ; Interleukin-8 ; metabolism ; Leukocyte Count ; Male ; Neutrophils ; cytology ; Respiratory System ; metabolism ; pathology ; Sputum ; metabolism
2.Cholesterol Depletion in Cell Membranes of Human Airway Epithelial Cells Suppresses MUC5AC Gene Expression.
Kee Jae SONG ; Na Hyun KIM ; Gi Bong LEE ; Ji Hoon KIM ; Jin Ho KWON ; Kyung Su KIM
Yonsei Medical Journal 2013;54(3):679-685
PURPOSE: If cholesterol in the cell membrane is depleted by treating cells with methyl-beta-cyclodextrin (MbetaCD), the activities of transmembrane receptors are altered in a cell-specific and/or receptor-specific manner. The proinflammatory cytokines, IL-1beta is potent inducers of MUC5AC mRNA and protein synthesis in human airway epithelial cells. Cells activated by IL-1beta showed increased phosphorylation of extracellular signal regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK). Thus, we investigated the effects of cholesterol depletion on the expression of MUC5AC in human airway epithelial cells and whether these alterations to MUC5AC expression were related to MAPK activity. MATERIALS AND METHODS: After NCI-H292 cells were pretreated with 1% MbetaCD before adding IL-1beta for 24 hours, MUC5AC mRNA expression was determined by reverse transcription-polymerase chain reaction (RT-PCR) and real time-PCR. Cholesterol depletion by MbetaCD was measured by modified microenzymatic fluorescence assay and filipin staining. The phosphorylation of IL-1 receptor, ERK and p38 MAPK, was analyzed by western blot. RESULTS: Cholesterol in the cell membrane was significantly depleted by treatment with MbetaCD on cells. IL-1beta-induced MUC5AC mRNA expression was decreased by MbetaCD and this decrease occurred IL-1-receptor-specifically. Moreover, we have shown that MbetaCD suppressed the activation of ERK1/2 and p38 MAPK in cells activated with IL-1beta. This result suggests that MbetaCD-mediated suppression of IL-1beta-induced MUC5AC mRNA operated via the ERK- and p38 MAPK-dependent pathway. CONCLUSION: Cholesterol depletion in NCI-H292 cell membrane may be considered an anti-hypersecretory method since it effectively inhibits mucus secretion of respiratory epithelial cells.
Cell Membrane/drug effects/*metabolism
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Cholesterol/*metabolism
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Epithelial Cells/metabolism
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Gene Expression
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Humans
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Mucin 5AC/genetics/*metabolism
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Respiratory System/*metabolism/pathology
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beta-Cyclodextrins/pharmacology
3.Effect of nuclear factor-kappaB on airway remodeling in asthmatic rats.
Shuyun XU ; Yongjian XU ; Zhenxiang ZHANG ; Wang NI ; Shixin CHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(1):13-18
In order to investigate the effect of nuclear factor-kappaB (NF-kappaB) on airway remodeling in asthmatic rats, 18 Wistar rats were divided into three groups: asthmatic group; pyrrolidine dithiocarbamate (PDTC) group, in which rats were injected intraperitoneally with NF-kappaB specific inhibitor PDTC (100 mg/kg) before ovalbumin (OVA) challenge; control group. The NF-kappaB activity and the expression of inhibitory protein kappaBalpha (I-kappaBalpha) in airway were detected by electrophoretic mobility shift assay (EMSA), Western blot and immunohistochemistry respectively. The infiltration of inflammatory cells, the number of Goblet cells, the area of collagen and smooth muscle in airway were measured by means of image analysis system. The results showed that with the up-regulation of airway NF-kappaB activity in asthmatic group, the number of goblet cells (3.084 +/- 0.86/100 microm basement membrane (BM)), the area of collagen (24.71 +/- 4.24 microm2/microm BM) and smooth muscle (13.81 +/- 2.11 microm2/microm BM) in airway were significantly increased (P<0.05) as compared with control group (0.14 +/- 0.05/100 microm BM, 14.31 +/- 3.16 microm2/microm BM and 7.67 +/- 2.35 microm2/microm BM respectively) and PDTC group (0.33 +/- 0.14/100 microm BM, 18.16 +/- 2.85 microm/microm BM and 8.95 +/- 2.16 microm2/microm BM respectively). However, there was no significant difference between PDTC group and control group (P>0.05). It was concluded that the activity of NF-kappaB is increased in airway of asthmatic rats. Inhibition of NF-kappaB activation can attenuate constructional changes in asthma airway, suggesting NF-kappaB may contribute to asthmatic airway remodeling.
Animals
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Asthma
;
metabolism
;
physiopathology
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Epithelium
;
metabolism
;
physiopathology
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I-kappa B Proteins
;
metabolism
;
Lung
;
pathology
;
physiopathology
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Mice
;
NF-kappa B
;
metabolism
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Ovalbumin
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Pyrrolidines
;
Rabbits
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Rats
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Rats, Wistar
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Respiratory System
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metabolism
;
physiopathology
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Thiocarbamates
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Transcription Factor RelA
4.The effect of budesonide on the eosinophil infiltration in airway of asthmatic rats.
Yi-ping LIN ; Ye HU ; Zhi-hua JIN
Chinese Journal of Applied Physiology 2009;25(3):365-367
Animals
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Anti-Inflammatory Agents
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pharmacology
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Asthma
;
pathology
;
physiopathology
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Budesonide
;
pharmacology
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Eosinophils
;
drug effects
;
pathology
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Inflammation
;
pathology
;
physiopathology
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NF-kappa B
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metabolism
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Random Allocation
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Rats
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Rats, Sprague-Dawley
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Respiratory System
;
pathology
5.Effect of MAPK signal transduction pathway inhibitor U0126 on aquaporin 4 expression in alveolar type II cells in rats with oleic acid-induced acute lung injury.
Chun-ling CHEN ; Tao-ping LI ; Li-hua ZHU
Journal of Southern Medical University 2009;29(8):1525-1528
OBJECTIVETo investigate the relationship between aquaporin 4 (AQP4) in alveolar type II (AT-II) cells and MAPK signaling pathway in rats with early-stage oleic acid-induced acute lung injury (ALI) and acute respiratory distress syndrome (ARDS).
METHODSThree groups of rats, namely the normal control, ALI and U0126 treatment group were used in this study. After oleic acid-induced ALI in the latter two groups, the rats in the treatment group received 100 micromol/L U0126 treatment at the dose of 10 micro, and dimethyl sulfoxide (DMSO) were given in the normal control and ALI groups. Arterial blood gas and the extravascular lung water (EVLW) content were measured after the treatments, and pathological changes in the lung tissues were observed microscopically. ATII cells were isolated from the lung tissues and identified using tannic acid staining and alkaline phosphatase (APK) staining. The expression of AQP-4 mRNA in the cells was detected with RT-PCR.
RESULTSBlood gas analysis, HE staining and EVLW content measurement revealed severer injury of the lung tissues in ALI group than in the normal control group, but the severity was comparable between the treatment and ALI groups. RT-PCR demonstrated significantly increased AQP-4 mRNA expression in ALI group as compared with that in the normal control group, and U0126 treatment resulted in obvious reduction in AQP-4 mRNA expression in the U0126 treatment group.
CONCLUSIONOleic acid-induced ALI results in the activation of MAPK signaling pathway and up-regulation of AQP-4 mRNA expression in the ATII cells of rats.
Acute Lung Injury ; chemically induced ; metabolism ; pathology ; Animals ; Aquaporin 4 ; genetics ; Butadienes ; pharmacology ; Extravascular Lung Water ; metabolism ; Gene Expression Regulation ; drug effects ; MAP Kinase Signaling System ; drug effects ; Male ; Nitriles ; pharmacology ; Oleic Acid ; pharmacology ; Pulmonary Alveoli ; enzymology ; metabolism ; pathology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Respiratory Distress Syndrome, Adult ; chemically induced ; metabolism ; pathology
6.Involvement of the p38 mitogen-activated protein kinase signal transduction pathway in burns-induced lung injury.
Xu-lin CHEN ; Zhao-fan XIA ; Duo WEI ; Yong-jie WANG ; Chang-rong WANG
Chinese Medical Journal 2005;118(4):329-332
Animals
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Body Water
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metabolism
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Capillary Permeability
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Hydrazones
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pharmacology
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Lung
;
metabolism
;
pathology
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MAP Kinase Signaling System
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Male
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Phosphorylation
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Rats
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Rats, Sprague-Dawley
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Respiratory Distress Syndrome, Adult
;
etiology
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p38 Mitogen-Activated Protein Kinases
;
physiology
7.Airway Inflammation and Allergen Specific IgE Production May Persist Longer Than Airway Hyperresponsiveness in Mice.
Yoon Seok CHANG ; Yoon Keun KIM ; Tae Bum KIM ; Hye Ryun KANG ; Sun Sin KIM ; Joon Woo BAHN ; Kyung Up MIN ; You Young KIM ; Sang Heon CHO
Journal of Korean Medical Science 2004;19(1):69-73
During the preclinical study of new therapeutic modality, we evaluate whether the treatment can reverse the established asthma phenotypes in animal model. However, few have reported on the long term persistence of asthma phenotypes upon re-challenge with allergen (secondary challenge) in animal model. We evaluated the persistence of asthma phenotypes by secondary challenge at different times in previously challenged murine asthma model. BALB/c mice sensitized by intraperitoneal injections of 20 microgram of ovalbumin and 1 mg of alum on days 1 and 14 were challenged initially by the inhalation of 1% ovalbumin for 30 min on days 21, 22, and 23. Each group of mice was rechallenged at 5, 7, 9, or 12 weeks after the initial challenge. Airway hyperresponsiveness, BAL fluid, airway histology and serum ovalbumin-specific IgE level were evaluated. Airway eosinophilia, airway inflammation and serum ovalbumin-specific IgE production persisted upon secondary allergen challenges at least 12 weeks after the initial challenge. However, airway hyperresponsiveness persisted only until mice were rechallenged 7 weeks after the initial challenge. Airway inflammation and allergen specific IgE production may persist longer than airway hyperresponsiveness in a mouse asthma model of secondary allergen challenge.
Allergens
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Animals
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Asthma/metabolism/*pathology
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Bronchial Hyperreactivity/*diagnosis
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Bronchoalveolar Lavage
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Bronchoalveolar Lavage Fluid
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Female
;
Immunoglobulin E/*biosynthesis/chemistry
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*Inflammation
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Lung/pathology
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Mice
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Mice, Inbred BALB C
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Ovalbumin/pharmacology
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Phenotype
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Respiratory Hypersensitivity/*diagnosis
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Respiratory System/pathology
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Support, Non-U.S. Gov't
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Time Factors
8.Two Cases of Citrullinemia Presented with Strokes.
Hyun Mi KIM ; Jae Bok KIM ; Jung Ho KIM ; Sang Jin BAE ; Chong Hyun YOON ; Han Wook YOO
Journal of the Korean Pediatric Society 1999;42(3):437-441
Urea cycle disorders are characterized by encephalopathy, respiratory alkalosis, and hyperammonemia. A urea cycle disorder should be considered a diagnostic possibility in any patient regardless of age with occult encephalopathy. The most common central nervous system pathology of urea cycle disorder is cerebral edema. The cerebral edema is caused by astrocyte swelling secondary to hyperammonemia and intracellular glutamine accumulation. Strokes in children occur in conjunction with cardiac disease, hematologic disorders, mitochondrial encephalopathy, trauma, intracranial infections and migraines. Recently, several inborn errors in metabolism have been recognized as possible causes of stroke. To our knowledge, there have been several reports on ornithine transcarbamylase deficiency with stroke. However, the case of citrullinemia presenting with a stroke-like episode has not been described previously. We report two infantile cases of citrullinemia with initial presentation of stroke. The differential diagnosis of unexplained strokes should include inborn errors of urea cycle metabolism during childhood.
Alkalosis, Respiratory
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Astrocytes
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Brain Edema
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Central Nervous System
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Child
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Citrullinemia*
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Diagnosis, Differential
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Glutamine
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Heart Diseases
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Humans
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Hyperammonemia
;
Metabolism
;
Migraine Disorders
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Mitochondrial Diseases
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Ornithine Carbamoyltransferase Deficiency Disease
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Pathology
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Stroke*
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Urea
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Urea Cycle Disorders, Inborn
9.Effect of all-trans retinoic acid on airway inflammation in asthmatic rats and its mechanism.
Hong FANG ; Hongfang JIN ; Hongwei WANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2004;24(3):229-232
The inhibitive effects of all-trans retinoic acid (ARTA) on airway inflammation in asthmatic rats and its mechanism on the basis of the regulation of nuclear factor kappaB (NF-kappaB) were explored. Thirty-two SD rats were randomly divided into 4 groups: control group, asthma group, dexamethasone treatment group and retinotic acid treatment group. The total and differential cell counts in the collected bronchoalveolar lavage fluid (BALF) were measured. The pathological changes in lung tissues were estimated by scoring. The expression of NF-kappaB inhibitor (IkappaBa), NF-kappaB, intercellular adhering molecule-1 (ICAM-1) in lung tissue was detected by immunohistochemical method. The results showed that in the two treatment groups, the total cell counts and proportion of inflammatory cells in BALF were significantly reduced, but there was no significant difference in differential cell counts in BALF between them. The pathological changes in lung tissues in the treatment groups were significantly attenuated as compared with asthma group. Except the epithelial injury in retinotic acid treatment group was milder than in dexamethasone treatment group, the remaining lesions showed no significant difference between them. In the two treatment groups, the expression of IkappaBa was increased, while the expression of NF-kappaB and ICAM-1 decreased with the difference between the two groups being not significant. It was concluded that the similar anti-inflammatory effects and mechanism of ATRA on airway in asthmatic rats to those of dexamethasone were contributed to the increase of cytoplasmic IkappaBa content and suppression of NF-kappaB activation and expression.
Animals
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Asthma
;
metabolism
;
pathology
;
Bronchoalveolar Lavage Fluid
;
cytology
;
Dexamethasone
;
pharmacology
;
I-kappa B Proteins
;
biosynthesis
;
genetics
;
Inflammation
;
Intercellular Adhesion Molecule-1
;
biosynthesis
;
genetics
;
Male
;
NF-kappa B
;
biosynthesis
;
genetics
;
Random Allocation
;
Rats
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Rats, Sprague-Dawley
;
Respiratory System
;
Tretinoin
;
pharmacology
10.Effect of spearmint oil on lipopolysaccharide induced emphysema-like changes and expression of matrix metalloproteinase-9.
Junbo LIU ; Yan WANG ; Fadi TANG ; Chenxi YU ; Mengshan HUANG ; Xiaojing ZHAO ; Youfa ZHU
China Journal of Chinese Materia Medica 2011;36(8):1054-1059
OBJECTIVETo investigate the effect of spearmint oil on emphysema-like changes and the expression of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta(IL-1beta), matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-9) in lipopolysaccharide (LPS) treated rats.
METHODEmphysematous changes model was induced by intratracheal instillation of LPS once a week for up to 8 weeks in rats. Rats were divided into control, dexamethasone (0.3 mg x kg(-1)), and spearmint oil (10, 30,100 mg x kg(-1)) groups. Each group was treated with saline, dexamethasone, and spearmint of oil respectively for 4 weeks. Then total and different white blood cell counts in bronchoalveolar lavage fluid(BALF) were carried out. The pathologic changes of lung tissue such as alveolar structure, airway inflammation, and goblet cell metaplasia were observed by HE and AB-PAS staining. Expression of TNF-alpha, IL-1beta, TIMP-1 and MMP-9 were measured.
RESULTBoth spearmint and dexamethasone decreased the destruction of pulmonary alveolus. The total and different white blood cell counts in BALF including neutrophile and lymphocyte of spearmint oil 100 mg x kg(-1) and dexamethasone group were significantly reduced, and the goblet cell metaplasia was also inhibited. Dexamethasone had inhibitory effect on the expression of TNF-alpha, IL-1beta, TIMP-1 and MMP-9. Spearmint oil 30, 100 mg x kg(-1) significantly reduced TNF-alpha and IL-1beta respectively. Spearmint oil 10, 30 and 100 mg x kg(-1) had no effect on the expression of TIMP-1, but could decrease the expression of MMP-9 significantly in lung tissues.
CONCLUSIONSpearmint oil has protective effect on rats with emphysematous changes, since it improves alveolar destruction, pulmonary inflammation, and goblet cell metaplasia. The mechanism may include reducing TNF-alpha, IL-1beta content and inhibiting overexpression of matrix metalloproteinase-9 in lung tissues.
Animals ; Azo Compounds ; pharmacology ; Bronchoalveolar Lavage Fluid ; cytology ; Goblet Cells ; drug effects ; Interleukin-1beta ; drug effects ; metabolism ; Leukocytes ; drug effects ; metabolism ; Lipopolysaccharides ; Lymphocytes ; drug effects ; metabolism ; Matrix Metalloproteinase 9 ; drug effects ; metabolism ; Mentha spicata ; chemistry ; Metaplasia ; Monocytes ; drug effects ; metabolism ; Neutrophils ; drug effects ; metabolism ; Phytotherapy ; Plant Oils ; therapeutic use ; Pulmonary Emphysema ; chemically induced ; drug therapy ; enzymology ; pathology ; Rats ; Respiratory System ; drug effects ; pathology ; Tissue Inhibitor of Metalloproteinase-1 ; drug effects ; metabolism ; Tumor Necrosis Factor-alpha ; drug effects ; metabolism