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Objective To study the protective effect of melatonin on PM2.5-induced inflammation and lymphangiogenesis in the lung of ApoE-/- mice. Methods Twenty-eight male ApoE-/- mice were randomized into # control group, PM2.5 group, melatonin group and PM2.5 + melatonin group. All mice were fed with western diet for 24 weeks. From the 25th week, mice in the melatonin group and PM2.5 + melatonin group were daily orally gavage with melatonin (20 mg/kg • bw) for 8 weeks; mice in the PM2.5 group and PM2.5 + melatonin group were exposed to PM2.5 by tracheal instillation (5mg/kg); and mice in the control group and melatonin group were instilled with saline at the same time. After 24 h of PM2.5 exposure, mice were euthanized and weight gain and lung weight/body weight ratio in four groups were analyzed. The concentrations of inflammatory cytokines (TNF - α and IL-6) in the lung tissue of mice were measured. Immunofluorescence staining of lung tissue was visualized the lymphatic marker LYVE1 expression. Western Blot was used to assess the protein expression levels of lymphangiogenesis markers PROX1 and LYVE1, lymphangiogenesis regulatory proteins VEGF-C and VEGFR-3 in lung tissues. Results The levels of TNF-α and IL-6 and the protein expressions of PROX1, LYVE1, VEGF-C and VEGFR-3 in lung tissues of PM2.5 group were significantly higher than the control group (P < 0.05). Moreover, the levels of TNF-α, IL-6 and the protein expressions of PROX1, LYVE1, VEGF-C and VEGFR-3 in lung tissues of the PM2.5 + melatonin group were significantly lower than the PM2.5 group (P < 0.05). Conclusion Ambient PM2. 5 exposure obviously increases lung inflammation of ApoE-/- mice, and may increase lymphangiogenesis in lung through regulating the VEGF-C/VEGFR-3 signaling pathway in the lung tissues; melatonin markedly improves PM2.5-induced lung inflammation and reduces lymphangiogenesis in lung. [ABSTRACT FROM AUTHOR]

Objective To identify the effective components and key molecules of Yiqi Huoxu compound(YQHX) in the treatment of pulmonary inflammation and fibrosis in scleroderma, providing basis for the development of drug targets for scleroderma. Methods A mouse model of pulmonary fibrosis was established by a single tracheal perfusion of bleomycin. Twenty-four mice were randomly divided into four groups, i.e. intragastric PBS, bleomycin alone, intragastric Yiqi Huoxue compound, and control. HE staining and Masson staining were used to verify the therapeutic effect of Yiqi Huoxue compound on lung inflammation and fibrosis. The active components and target genes of anti-fiber salvia miltiorrhiza were obtained from TCMSP database. PPI network, GO/KEGG enrichment analysis, molecular docking and other bioinformatics analyses were integrated to identify the main active ingredients and key targets of salvia miltiorrhiza in the treatment of scleroderma, and to analyze the relevant signaling pathways. The expression levels of CD3 antibody, p-PI3K antibody and p-AKT antibody were analyzed by immunohistochemistry to verify the effects of Yiqi Huoxu compound on inflammation and key pathways. Results The results of HE staining and Masson staining showed that the severity of fibrosis and inflammatory infiltration were decreased, suggesting that Yiqi Huoxue compound could significantly improve the pulmonary fibrosis, inflammation and tissue damages. The key component of salvia miltiorrhiza was mainly luteolin. PPI network analysis showed that AKT1, EGFR, ESR1, TNF and IL6 were the core targets of salvia miltiorrhiza in the treatment of scleroderma. KEGG enrichment analysis showed that the genes were mainly enriched in the PI3K-AKT pathway. Molecular docking analysis showed the potential binding sites between luteolin and four key targets. The immunohistochemical results showed that YQHX significantly decreased bleomycin-induced CD3+T cell infiltration, p-PI3K and p-AKT levels in mouse lung tissue. Conclusions YQHX has significant therapeutic effects on lung inflammation and fibrosis. Luteolin may be a key effective molecule in YQHX, and treat scleroderma lung inflammation and fibrosis by targeting the PI3K-AKT pathway.

Objective To identify the effective components and key molecules of Yiqi Huoxu compound (YQHX) in the treatment of pulmonary inflammation and fibrosis in scleroderma, providing basis for the development of drug targets for scleroderma. Methods A mouse model of pulmonary fibrosis was established by a single tracheal perfusion of bleomycin. Twenty-four mice were randomly divided into four groups, i. e. intragastric PBS, bleomycin alone, intragastric Yiqi Huoxue compound, and control. HE staining and Masson staining were used to verify the therapeutic effect of Yiqi Huoxue compound on lung inflammation and fibrosis. The active components and target genes of anti-fiber salvia miltiorrhiza were obtained from TCMSP database. PPI network, GO/KEGG enrichment analysis, molecular docking and other bioinformatics analyses were integrated to identify the main active ingredients and key targets of salvia miltiorrhiza in the treatment of scleroderma, and to analyze the relevant signaling pathways. The expression levels of CD3 antibody, p-PI3K antibody and p-AKT antibody were analyzed by immunohistochemistry to verify the effects of Yiqi Huoxu compound on inflammation and key pathways. Results The results of HE staining and Masson staining showed that the severity of fibrosis and inflammatory infiltration were decreased, suggesting that Yiqi Huoxue compound could significantly improve the pulmonary fibrosis, inflammation and tissue damages. The key component of salvia miltiorrhiza was mainly luteolin. PPI network analysis showed that AKT1, EGFR, ESR1, TNF and II6 were the core targets of salvia miltiorrhiza in the treatment of scleroderma. KEGG enrichment analysis showed that the genes were mainly enriched in the PI3K-AKT pathway. Molecular docking analysis showed the potential binding sites between luteolin and four key targets. The immunohistochemical results showed that YQHX significantly decreased bleomycin-induced CD3 + T cell infiltration, p-PI3K and p-AKT levels in mouse lung tissue. Conclusions YQHX has significant therapeutic effects on lung inflammation and fibrosis. Luteolin may be a key effective molecule in YQHX, and treat scleroderma lung inflammation and fibrosis by targeting the PI3K-AKT pathway. [ABSTRACT FROM AUTHOR]

[Objectives]This study aimed to investigate the effect of the pulmonary microbiota on PM2.5-induced lung inflammatory injury in broilers and to explore the role of Enterococcus cecorum on PM2.5-induced lung inflammatory injury using E.cecorum-treated alveolar epithelial cells. [Methods]A total of fourty-five male AA broilers(19-day-old)were randomly divided into three groups:the control group, the PM2.5 group(PM), and the pulmonary microbiota-intervened group(ABX-PM). From 21 days of age, broilers in the ABX-PM group were intratracheally instilled with antibiotics once a day for three days. Meanwhile, broilers in the other two groups were simultaneously instilled with sterile saline. At 24 and 26 days of age, broilers in the PM and ABX-PM groups were intratracheally instilled with PM2.5 suspension to induce lung inflammatory injury, and broilers in the control group were simultaneously instilled with sterile saline. Twenty-four hours after the last intratracheal instillation of PM2.5, we weighed the broilers and collected lung tissue, bronchoalveolar lavage fluid(BALF), and serum samples. We measured the levels of inflammatory factors in BALF and serum. Besides, we measured the gene expression levels of tight junctions(TJ)protein, the relative content of Enterococcus spp. and E.cecorum in the lung tissue, and the correlation between the relative content of E.cecorum and inflammatory factor levels. In addition, type Ⅱ alveolar epithelial cells(A549)were cultured and divided into two groups. The control group was treated with DMEM/F12 medium without E.cecorum, while the E.cecorum treatment group was treated with DMEM/F12 medium containing 1×104 CFU·mL-1 E.cecorum. We compared the gene expression levels of inflammatory factors, TJ, and apoptosis between the two groups after 6 h of treatment. [Results]The levels of IL-8 in BALF and serum, as well as the mRNA expression levels of occludin and claudin-1 in lung tissue, were significantly higher in the PM group than those in the control group. The claudin-5 mRNA expression level was significantly lower in the PM group than the control group, but the above indicators were not statistically different in the ABX-PM group compared to the control group. The relative contents of Enterococcus spp. and E.cecorum in the PM group were significantly higher than those in the control and ABX-PM groups. The relative content of E.cecorum was significantly positively correlated with the levels of IL-1β, IL-6, and IL-8 in BALF and IL-1β in serum. In the cellular experiment, the mRNA expression levels of tnf-α, il-1β, il-6, and il-8 were significantly higher, while the mRNA expression level of il-10 was significantly lower in the E.cecorum treatment group than those in the control group. The mRNA expression levels of zo-1 and claudin-1 in the E.cecorum treatment group were significantly higher than those in the control group. The ratio of bax/bcl-2 mRNA expression levels and the mRNA expression level of caspase-3 were significantly higher, while the mRNA expression level of bcl-2 was significantly lower in the E.cecorum treatment group than those in the control group. [Conclusions]The pulmonary microbiota intervention reduced PM2.5-induced proliferation of E.cecorum, and alleviated the degree of lung inflammatory injury in broilers, indicating the involvement of pulmonary microbiota in the process of PM2.5-induced lung inflammation injury in broilers. Moreover, E.cecorum could induce an inflammatory response in alveolar epithelial cells, which suggested that the imbalance of pulmonary microbiota aggravated lung inflammation injury. [ABSTRACT FROM AUTHOR]

Objective: To investigate the roles of angiotensin-converting enzyme 2 (ACE2) in ozone-induced pulmonary inflammation and airway remodeling in mice., Methods: Sixteen wild-type (WT) C57BL/6J mice and 16 ACE2 knock-out (KO) mice were exposed to either filtered air or ozone (0.8 ppm) for 3 h per day for 5 consecutive days. Masson's staining and HE staining were used to observe lung pathologies. Bronchoalveolar lavage fluid (BALF) was collected and the total cell count was determined. The total proteins and cytokines in BALF were determined by BCA and ELISA method. The transcription levels of airway remodeling-related indicators in the lung tissues were detected using real-time quantitative PCR. The airway resistance of the mice was measured using a small animal ventilator with methacholine stimulation., Results: Following ozoneexposure ACE2 KO mice had significantly higher lung pathological scores than WT mice ( P < 0.05). Masson staining results showed that compared with ozone-exposed WT mice, ozone-exposed ACE2 KO mice presented with significantly larger area of collagen deposition in the bronchi [(19.62±3.16)% vs (6.49±1.34)%, P < 0.05] and alveoli [(21.63±3.78)% vs (4.44±0.99)%, P < 0.05]. The total cell count and total protein contents in the BALF were both higher in ozone-exposed ACE2 KO mice than in WT mice, but these differences were not statistically significant ( P > 0.05). The concentrations of IL-6, IL-1β, TNF- α , CXCL1/KC and MCP-1 in the BALF were all higher in ozone-exposed ACE2 KO mice than in ozone-exposed WT mice, but only the difference in IL-1β was statistically significant ( P < 0.05). The transcription levels of MMP-9, MMP-13, TIMP 4, COL1A1, and TGF-β in the lung tissues were all significantly higher in ozone-exposed ACE2 KO mice ( P < 0.01). No significant difference was found in airway resistance between ozone-exposed ACE KO mice and WT mice after challenge with 0, 10, 25, or 100 mg/mL of methacholine., Conclusion: ACE2 participates in ozone-induced lung inflammation and airway remodeling in mice.