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Cysathionine γ-lyase (CSE) is a core enzyme for the synthesis of endogenous H
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Objective To investigate the therapeutic effect and possible mechanism of adenosine A2A receptor agonist (CGS21680) combined with bone marrow mesenchymal stem cells (BMMSC) transplantation in acute liver failure (ALF).Methods Fifty male C57BL/6 mice, 6-8 weeks old, were fed with standard diet for 1 week and randomly divided into 5 groups according to random number table: healthy control group (n=6), model group (n=11), BMMSC group (n=11), CGS21680/BMMSC group (n=11) and CGS21680 group (n=11).Except healthy control group, the other mice were injected with D-GalN and lipopolysaccharide (LPS) to establish ALF model.Ten hours later, CGS21680/BMMSC group and CGS21680 group were injected intraperitoneally with adenosine A2A receptor agonist CGS21680 (2.1 mg/kg).In addition, the BMMSC group and CGS21680/BMMSC group were injected BMMSC (1×10.6) through tail vein.After 24 hours, pathological changes of liver tissue was observed by hematoxylin and eosin staining.The change of proportion of mouse splenic Treg among CD4+T lymphocytes was detected by flow cytometry.Toll-like receptor (TLR)4 and nuclear factor (NF)-κB expression levels in liver tissue were detected by real-time fluorescence quantitative polymerase chain reaction (PCR) and Western blot.One-way analysis of variance (one-way ANOVA) and SNK-q test was conducted for data analysis.Results Serum IL-6 levels were (23.67±2.97) pg/mL in healthy control group, (151.47±6.03) pg/mL in model control group, (72.10±3.74) pg/mL in BMMSC group, (53.35±2.50) pg/mL in CGS21680/BMMSC group and (84.85±3.25) pg/mL in CGS21680 group.The differences between healthy control group and the other 4 groups were all statistically significant (t=46.02, 25.51, 19.58 and 34.03, respectively, all P<0.01).Serum TNF-ɑ levels were (24.62±3.19) pg/mL in healthy control group, (102.25±2.10) pg/mL in model control group, (54.71±2.23) pg/mL in BMMSC group, (42.20±4.72) pg/mL in CGS21680/BMMSC group and (81.76±3.50) pg/mL in CGS21680 group.The differences between healthy control group and the other 4 groups were all statistically significant (t=46.49, 19.97, 7.72 and 29.57, respectively, all P<0.01).The differences of spleen Treg proportion in healthy control group were statistically significant compared with model control group, BMMSC group, CGS21680/BMMSC group and CGS21680 group (t=51.67, 12.22, 5.91 and 18.21, respectively, all P<0.01).The differences of TLR4 mRNA levels of liver tissue in healthy control group were statistically significant compared with model control group, BMMSC group, CGS21680/BMMSC group and CGS21680 group (t=26.31, 21.33, 13.24 and 27.14, respectively, all P<0.05).The differences of NF-κB mRNA level of liver tissue in healthy control group were statistically significant compared with model control group, BMMSC group, CGS21680/BMMSC group and CGS21680 group (t=16.56, 16.34, 7.83 and 13.11, respectively, all P<0.05).The differences of TLR4 protein level in liver tissue of healthy control group were statistically significant compared with model control group, BMMSC group, CGS21680/BMMSC group and CGS21680 group (t=35.60, 10.38, 6.05 and 18.02, respectively, all P<0.05).The differences of liver NF-κB protein level in the healthy control group were statistically significant compared with model control group, BMMSC group, CGS21680/BMMSC group and CGS21680 group (t=10.80, 7.30, 4.61 and 13.24, respectively, all P<0.05).Conclusions Adenosine A2A receptor agonist combined with BMMSC can significantly up-regulate the proportion of Treg cells in acute liver failure mice and inhibit the TLR4/NF-κB pathway activation, with both coordinated regulation, and further inhibit the liver inflammation.
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Objective To explore the mechanism of azithromycin for intervening with inflammation of rats with chronic ob‐structive pulmonary disease (COPD) via TLR‐4/NF‐κB signal pathway .Methods Thirty‐six SD rats were divided into health con‐trol group ,model group and azithromycin group .The rat COPD model in the model group and the azithromycin group was estab‐lished by smoking and intratracheal administration of lipopolysaccharide for 1 month .At 30 min before smoking ,the azithromycin group was given azithromycin 50 mg/kg by gavage ,while the health control group and model group were given the equal amount of normal saline .One month later ,rats were sacrificed and lung tissue was obtained .The pathological morphology of the lung was ob‐served by the HE staining .The enzyme linked immunosorbent assay (ELISA) method was used to detect the level of TNF‐α,IL‐1βand IL‐6 from lung tissue homogenate .The expression of TLR4 ,NF‐κB and pp65 mRNA was detected by RT‐PCR .The expression of TLR4 ,NF‐κB ,pp65 ,MMP‐2 and MMP‐9 protein was detected by Western blot .The cytological classification and count in bron‐choalveolar lavage fluid (BALF) were performed .Results Compared with the model group ,the azithromycin group could improve the lung tissue morphology ,decreased the neutrophil granulocyte count(P< 0 .01) ,reduced the secretion of TNF‐α,IL‐1β,IL‐6 , MMP‐2 and MMP‐9 in lung tissue homogenates(P<0 .01) ,and suppressed the expression of TLR4 and pp65 phosphorylation level (P<0 .01) .Conclusion Azithromycin can intervene with inflammation of rats with COPD .