ObjectiveTo investigate the role and mechanism of S100A8/A9 in rat model of chronic obstructive pulmonary disease (COPD). Methods Twelve Wistar rats were randomly divided into a normal control group and a COPD group. The COPD model was established by exposing the rats to cigarette smoke and injected lipopolysaccharide (LPS) in bronchus for 1 month. The pathological changes of the lung tissue were observed under light microscope, and the emphysema indexes of pulmonary mean linear intercept (MLI), mean alveolar numbers (MAN) and pulmonary alveolar area (PAA) were analyzed by image analysis system. The concentrations of S100A8/A9 in serum and bronchoalveolar lavage fluid (BALF) were measured by enzyme linked immunosorbent assay. The mRNA expression levels of S100A8, S100A9, Toll-like receptor-4 (TLR4) and myeloid differentiation factor 88 (MyD88) of lung tissues were measured by real time polymerase chain reaction. The protein expressions of S100A8/A9, TLR4 and MyD88 of lung tissues were detected by immunohistochemistry. Results After cigarette smoking and LPS injection for 1 month, the rat lung tissue appeared in accordance with the typical pathological changes of COPD. The MLI, MAN and PAA had obvious difference compared with the normal control group (P<0.05). The concentrations of S100A8/A9 protein in BALF and serum of the COPD group were obviously higher than those of the normal control group (P<0.05). The levels of S100A8, S100A9, TLR4 and MyD88 mRNA of lung tissues in the COPD group were obviously higher than those in the normal control group (P<0.05), and the expression levels of S100A8 and S100A9 mRNA were positively correlated with the expression levels of TLR4 and MyD88 mRNA respectively (P<0.05). The levels of S100A8/A9, TLR4 and MyD88 protein of lung tissues in COPD group were obviously higher than those in normal control group (P<0.05), and the levels of S100A8/A9 protein were positively correlated with the levels of MyD88 and TLR4 protein (P<0.05). Conclusions As a new inflammatory mediator, S100A8/A9 may be involved in the occurrence and development of COPD. By up-regulating the expression of TLR4 and MyD88, the classical TLR4-MyD88 inflammatory pathway is activated, thus promotes the occurrence and development of COPD.
Objective To investigate the preventive effect of simvastatin，a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor，on hypoxic pulmonary hypertension and the relation between it and the angiotensin Ⅱ receptor-1（AT1R） expression in pulmonary arteriole.Methods Thirty male Sprague-Drawley rats were randomly allocated into three groups：a control group，a hypoxic group and a simvastatin preventive group.The animal model of hypoxic pulmonary hypertension was established by exposing the rats to normobaric hypoxic condition（8 h×6 d×3 w），and the preventive group were treated with simvastatin 10 mg/kg before hypoxic processing while the control and hypoxic groups were treated with sodium chloride.The mean pulmonary pressure（mPAP），serum cholesterol concentration，right ventricular hypertrophy index [RV/（LV+S）]，percentage of the wall thickness in the external diameter（WT%），percentage of the wall area in the total vascular area（WA%），and the AT1R expression in pulmonary arterioles were measured.Results When compared with the hypoxic group，in the preventive group，the mPAP and RV/（LV+S）obviously reduced [（22.6±3.86）mm Hg vs （29.3±2.27）mm Hg，（25.13±0.75）% vs （33.18±1.58）%，Plt;0.01 respectively]，the indices of wall thickness of rat pulmonary arteriole and area also decreased significantly [WT%：（15.98±1.96）% vs （25.14±1.85）%；WA%：（54.60±3.94）% vs 74.77±4.52）%；Plt;0.01 respectively]，and the positive degree of AT1R still lessened noticeably（1.23±0.09 vs 1.57±0.13，Plt;0.01）.All of the indices above in the hypoxic group increased markedly compared with the control group（Plt;0.01 respectively）.However，the differences of serum cholesterol among three groups were not significant（Pgt;0.05）.Conclusions Simvastatin can suppress the expression of AT1R in pulmonary vessel and prevent hypoxic pulmonary hypertension.
ObjectiveTo investigate the effect of maresin-1 (MaR1) on lung inflammation and MAPK signaling pathway in asthmatic mice.MethodsTwenty-four female BALB/c mice were randomly divided into normal group, asthma model group, MaR1 group and dexamethasone group. The asthma model was successfully established by using ovalbumin (OVA) combined with aluminum hydroxide, and then MaR1 and dexamethasone were respectively given to asthmatic mice. Serum, bronchoalveolar lavage fluid (BALF) and lung tissue were collected for further analysis. Pathological changes of lung tissue in mice were detected by hematoxylin-eosin and periodic acid-Schi?. Proportion of inflammatory cells in BALF classified by Swiss-Giemsa staining. Th2-related inflammatory cytokines, interleukin (IL)-4, IL-5, IL-13 and IgE in serum and BALF were detected by enzyme linked immunosorbent assay. The protein concentration of p-p38 and p-JNK in lung tissues were detected by Western blot.ResultsCompared with the normal group, the asthma model group had increased both airway inflammation and the number of goblet cells significantly (P<0.05). The number of various inflammatory cells in BALF had also increased significantly (P<0.05). The levels of IL-4, IL-5 and IL-13 in BALF and IgE and OVA-specific-IgE in serum were significantly increased (P<0.05). The protein contents of p-p38 and p-JNK in lung tissues were significantly increased (P<0.05). Compared with the asthma model group, both MaR1 and dexamethasone group had reduced inflammation and mucus secretion in lung tissue, number of inflammatory cells in BALF (P<0.05), levels of related inflammatory cytokines in BALF and IgE in serum (P<0.05), and expression of p-p38 and p-JNK proteins in lung tissue (P<0.05).ConclusionsMaR1 can inhibit the production and release of both Th2-related inflammatory cytokines and IgE, effectively reduce the inflammatory response and mucus production in lung tissues of asthmatic mice, with similar effect to dexamethasone. The mechanism may be related to the down-regulation of MAPKs signaling pathway.
Objective To observe the expression of S100A8 and S100A9 in alveolar macrophages (AMs) of chronic obstructive pulmonary disease (COPD) rats, and explore the effect on the release of inflammatory mediators from AMs in COPD rats. Methods Twelve adult male Wistar rats were randomly divided into a normal control group and a COPD group. The COPD model was established by exposing the rats to cigarette smoke and intratracheal injection of endotoxin for 1 month. The pathological changes of lung tissue of rats were observed under light microscope. Total cells counts and the number of AMs, lymphocytes, neutrophils in bronchoalveolar lavage fluid (BALF) of two groups were examined by Wright's staining methods. Rat AMs from the control group and the COPD group were isolated and cultured, and then treated with different doses of S100A8 and S100A9 for 6 hours and 12 hours. The levels of interleukin (IL)-8, IL-6 and tumour necrosis factor-α (TNF-α) in the AMs supernatants were measured by enzyme linked immunosorbent assay. The expression of S100A8 and S100A9 mRNA in AMs of rats were observed by in situ hybridization. The immunohistochemical method was used to observed the expression of S100A8/A9 protein of AMs. Results After cigarette smoking combined with intratracheal injection of endotoxin for 1 month, the lung tissue of rats showed typical pathological changes of COPD. Total cell counts and the number of AMs, lymphocytes, neutrophils in BALF of the COPD rats were significantly higher than those of the normal rats (P<0.05). Among them, the increase in the number of AMs was the most obvious. Compared with the control group, the expression of S100A8 mRNA, S100A9 mRNA and S100A8/A9 protein in AMs of the COPD group were up-regulated significantly (P<0.05). After the AMs of COPD rats were treated with S100A8 and S100A9, the contents of IL-8, IL-6 and TNF-α in AMs supernatants increased significantly in a time- and dose-dependent manner. When the AMs were treated with the same dose of S100A8 and S100A9 for the same time, the levels of IL-8, IL-6 and TNF-α in the AMs supernatant of the COPD group were higher than those of the normal control group. Conclusions The expression of S100A8 and S100A9 in cultured COPD rat AMs is significantly increased. S100A8 and S100A9 can promote the secretion and release of inflammatory factors IL-6, IL-8 and TNF-α from AMs of COPD rats in a time and dose-dependent manner. The effects of S100A8 and S100A9 on the secretion of IL-6, IL-8 and TNF-α in AM of COPD rats are significantly enhanced compared with those of normal rats.
