This study aims to explore the mechanism of fresh Phragmitis Rhizoma against chronic bronchitis airway inflammation. The SD rats of SPF grade were divided into control group, model group, Guilongkechuanning group(GLKCN, 1.125 g·kg~(-1)), high-dose fresh Phragmitis Rhizoma group(LG-HD, 15 g·kg~(-1)), and low-dose fresh Phragmitis Rhizoma group(LG-LD, 7.5 g·kg~(-1)). The chronic bronchitis models of rats in other groups except the control group were induced by the modified smoking method. From the 15 th day of modeling, the rats were given corresponding agents by gavage for 20 consecutive days. After the last administration, the rats were sacrificed for sample collection. Enzyme-linked immunosorbent assay(ELISA) was employed to detect serum transforming growth factor-β(TGF-β) and interleukin-6(IL-6) levels. The protein expression of TGF-β, IL-1β and IL-6 in lung tissue was detected by immunohistochemical method. Masson staining was performed to detect collagen fibers and muscle fibers in lung tissue, and HE staining to detect the pathological changes of lung tissue. Human bronchial epithelial(16 HBE) cells were cultured in vitro, and CCK-8(cell counting kit-8) method was used to detect the cytotoxicity of cigarette smoke extract(CSE) and fresh Phragmitis Rhizoma. After the exposure of 16 HBE cells to 3.5% CSE and appropriate concentration(800, 400 μg·mL~(-1)) of fresh Phragmitis Rhizoma for 24 h, quantitative real-time PCR was conducted to determine the mRNA levels of TGF-β and IL-1β, and Western blot was employed to determine the protein levels of TGF-β and IL-6 in the cells. The rat model of chronic bronchitis induced by smoking was successfully established. Fresh Phragmitis Rhizoma reduced serum TGF-β and IL-6 levels, down-regulated the protein levels of TGF-β, IL-1β, and IL-6 in lung tissue, and alleviated pathological changes and fibrotic lesions in lung tissue. Moreover, it down-regulated the CSE-induced protein expression of TGF-β and IL-6 as well as the mRNA level of TGF-β in 16 HBE cells. These results indicated that fresh Phragmitis Rhizoma could prevent airway inflammation from chronic bronchitis and promote cell repair by inhibiting the TGF-β signaling pathway.
Animals ; Bronchitis, Chronic/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Inflammation ; Lung ; Poaceae/chemistry* ; Rats ; Rats, Sprague-Dawley ; Rhizome ; Signal Transduction ; Transforming Growth Factor beta/genetics*

OBJECTIVE: To determine the relation between resistance of hypermutable Pseudomonas aeruginosa and beta-lactamases produced. METHODS: The bacteria cultured were identified with API 20NE system. Susceptibilities of the bacteria were detected by disk diffusion method. The hypermutable strains were tested with broth dilution assays. The beta-lactamases produced by these strains were characterized by 3-dimensional test and 2-mercaptopropanoic acid inhibited assays. RESULTS: Altogether 120 strains were analyzed and 45 (37.5%) trains were hypermutable.The resistant rates of hypermutable strains were close to or above 60.0% for imipenem, meropenem, cefoperazone/sulbactam, piperacillin/ tazobactam, ceftazidime, cephfime, aztreonam, amikacin and ciprofloxacin.The 3-dimensional test showed that 18 (40.0%) strains produced extended spectrum beta-lactamases (ESBLs), 25 (55.6%) strains produced AmpC enzymes, and 6 (13.3%) strains produced metallo-beta-lactamases. CONCLUSION The resistant rates of hypermutablce strains of Pseudomonas aeruginosa to routine antibiotics are high, which is one of the most important reasons for multi-drug resistance that the hypermutable strains produced ESBLs, AmpC enzymes, and metallo-beta-lactamases.
Adult ; Aged ; Aged, 80 and over ; Bronchitis, Chronic ; microbiology ; Drug Resistance, Multiple, Bacterial ; genetics ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Pseudomonas aeruginosa ; drug effects ; enzymology ; genetics ; Pulmonary Heart Disease ; microbiology ; beta-Lactamases ; biosynthesis

OBJECTIVETo explore the mechanism of anti-inflammatory effect of mangiferin.

METHODThe model of chronic bronchitis in rat was established by LPS + smoke. The activity of SOD, content of MDA and NO in BALF and serum, content of TNF-alpha and IL-8 were determined. The expression of RAW264.7 macrophage COX-2 mRNA induced by LPS in mice was detected by RT-PCR.

RESULTThe activity of SOD, the content of NO in BALF and serum in rat with chronic bronchitis were significantly higher with high, medium and low-dose of lg mangiferin (400,200,100 mg x kg(-1)), while the content of MDA, and the content of TNF-alpha and IL-8 in lung tissues were lower. The expression of RAW264.7 macrophage COX-2 mRNA induced by LPS was significantly reduced by mangiferin with 200,100, 50 micromol x L(-1).

CONCLUSIONThe anti-inflammatory mechanism of mangiferin is to relieve inflammation by raising the activity of SOD and content of NO and reducing the content of MDA and the expression of TNF-alpha, IL-8 and COX-2 mRNA.

Animals ; Bronchitis, Chronic ; drug therapy ; genetics ; immunology ; Cell Line ; Cyclooxygenase 2 ; genetics ; immunology ; Cytokines ; genetics ; immunology ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Gene Expression ; drug effects ; Humans ; Macrophages ; drug effects ; enzymology ; immunology ; Male ; Mice ; Rats ; Rats, Sprague-Dawley ; Xanthones ; administration & dosage ; pharmacology

This study is to investigate the protective effect of mangiferin on NF-kappaB (P65) and IkappaBalpha expression in peripheral blood mononuclear cell (PBMC) in rats with cigarette smoke induced chronic bronchitis. The rat model with chronic bronchitis was established by cigarette smoke. Real-time fluorescence RT-PCR was executed for evaluating the NF-kappaB (P65) and IKkappaBalpha gene expression in mononuclear cell, and flow cytometry for their protein expression. The serum hs-CRP (high-sensitivity C-reactive proteins) and TNF-alpha (tumor necrosis factor-alpha) were detected by enzyme-linked immunosorbent assay. The histopathological score was obtained from lung tissue HE staining slides of lung tissue. The results showed that mangiferin could markedly suppress the NF-kappaB (P65) mRNA and protein expression in mononuclear cell, while promote the IkappaBalpha mRNA and protein expression. Furthermore, mangiferin could lower serum hs-CRP and TNF-alpha level, and reduce the chronic inflammatory damage of bronchiole. These results suggested that mangiferin could notably ameliorate chronic bronchiole inflammation induced by cigarette smoke, and this protective effect might be linked to the regulation of NF-kappaB (P65) and IkappaBalpha expression in mononuclear cell.
Animals ; Bronchi ; pathology ; Bronchitis, Chronic ; blood ; etiology ; metabolism ; pathology ; C-Reactive Protein ; metabolism ; I-kappa B Kinase ; genetics ; metabolism ; Leukocytes, Mononuclear ; metabolism ; pathology ; Male ; Mangifera ; chemistry ; Plants, Medicinal ; chemistry ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tobacco Smoke Pollution ; Transcription Factor RelA ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; blood ; Xanthones ; isolation & purification ; pharmacology