Toxoplasma ROP16I/III ameliorated inflammatory bowel diseases via inducing M2 phenotype of macrophages.

BACKGROUND: Inflammatory bowel disease (IBD) is characterized by chronic and non-specific inflammation of the intestinal mucosa and mainly includes ulcerative colitis and Crohn's disease. AIM: To explore the beneficial effect of ToxoROP16I/III-induced M2 phynotype macrophages in homeostasis of IBDs through downregulation of M1 inflammatory cells. METHODS: RAW264.7 macrophages stimulated by lipopolysaccharide (LPS) (M1 cells) were co-cultured with Caco-2 cells as an inflammatory model of IBD in vitro. The expression of ToxoROP16I/III was observed in RAW264.7 macrophages that were transfected with pEGFP-rop16 I/III. The phenotypes of M2 and M1 macrophage cells were assessed by quantitative real-time reverse transcriptase polymerase chain reaction and the expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-6, transforming growth factor (TGF)-ß1, IL-10, inducible nitric oxide synthase (iNOS), and arginase-1 (Arg-1) was detected. The expression of iNOS, Arg-1, signal transducer and activator of transcription 3 (Stat3), p-Stat3, Stat6, p-Stat6, programmed death ligand-2 (PD-L2), caspase-3, -8, and -9 was analyzed by Western blotting, and Griess assays were performed to detect nitric oxide (NO). TNF-α, IL-1ß, IL-6, TGF-ß1, and IL-10 expression in the supernatants was detected by enzyme-linked immunosorbent assay, and Caco-2 cell apoptosis was determined by flow cytometry after mixing M1 cells with M2 cells in a Caco-2 cell co-culture system. RESULTS: M1 cells exhibited significantly increased production of iNOS, NO, TNF-α, IL-1ß, and IL-6, while ToxoROP16I/III induced macrophage bias to M2 cells in vitro, showing increased expression of Arg-1, IL-10 and TGF-ß1 and elevated production of p-Stat3 and p-Stat6. The mixed M1 and M2 cell culture induced by ToxoROP16I/III exhibited decreased production of NO and iNOS and upregulated expression of Arg-1 and PD-L2. Accordingly, Caco-2 cells became apoptotic, and apoptosis-associated proteins such as caspase-3, -8 and -9 were dampened during co-culture of M1 and M2 cells. Flow cytometry analysis showed that co-culture of M1 cells with Caco-2 cells facilitated the apoptosis of Caco-2 cells, but co-culture of M1 and M2 cells alleviated Caco-2 cell apoptosis. CONCLUSION: ToxoROP16I/III-induced M2 macrophages inhibited apoptosis of Caco-2 cells caused by M1 macrophages. This finding may help gain a better understanding of the underlying mechanism and represent a promising therapeutic strategy for IBDs.

[Characteristics of CO2 emission from Carex-dominated wetland in Poyang Lake in non-flooded period].

By using static chamber/gas chromatography, the CO2 fluxes in a Carex cinerascen-dominated wetland in the Poyang Lake Nanji Wetland National Nature Reserve were measured in nonflooded period (from September 2009 to April 2010). Two treatments were installed, i. e. , soil-plant system (TC) and aboveground plant removal (TJ), representing ecosystem respiration and soil respiration, respectively. There was an obvious seasonal variation in the ecosystem respiration and soil respiration. The respiration rate in treatment TC ranged from 89.57 to 1243.99 mg CO2 x m(-2) x h(-1), and that in TJ was from 75.30 to 960.94 mg CO2 x m(-2) x h(-1). Soil respiration accounted for 39% -84% of ecosystem respiration, with an average of 64%. Soil temperature was the main factor controlling the ecosystem respiration and soil respiration, explaining more than 80% of the respiration variance. The temperature coefficient (Q10), an index of temperature sensitivity for respiration, was 3.31 for ecosystem respiration and 2.75 for soil respiration. The Q10 value was higher in winter than in autumn and spring. No significant correlation was observed between soil moisture and CO2 fluxes. In non-flooded period, the C. cinerascens-dominated wetland acted as a carbon sink of atmospheric CO2, with a carbon uptake of 1717.72 g C x m(-2).