Schistosoma japonicum cystatin alleviates paraquat poisoning caused acute lung injury in mice through activating regulatory macrophages.

BACKGROUND: Paraquat (PQ) is a widely used herbicide that poisons human by accident or intentional ingestion. PQ poisoning causes systemic inflammatory response syndrome (SIRS) resulting in acute lung injury (ALI) with an extremely high mortality rate. Blood trematode Schistosoma japonicum-produced cystatin (Sj-Cys) is a strong immunomodulatory protein that has been experimentally used to treat inflammation related diseases. In this study, Sj-Cys recombinant protein (rSj-Cys) was used to treat PQ-induced lung injury and the immunological mechanism underlying the therapeutic effect was investigated. METHODS: PQ-induced acute lung injury mouse model was established by intraperitoneally injection of 20 mg/kg of paraquat. The poisoned mice were treated with rSj-Cys and the survival rate was observed up to 7 days compared with the group without treatment. The pathological changes of PQ-induced lung injury were observed by examining the histochemical sections of affected lung tissue and the wet to dry ratio of lung as a parameter for inflammation and edema. The levels of the inflammation related cytokines IL-6 and TNF-α and regulatory cytokines IL-10 and TGF-ß were measured in sera and in affected lung tissue using ELISA and their mRNA levels in lung tissue using RT-PCR. The macrophages expressing iNOS were determined as M1 and those expressing Arg-1 as M2 macrophages. The effect of rSj-Cys on the transformation of inflammatory M1 to regulatory M2 macrophages was measured in affected lung tissue in vivo (EKISA and RT-PCR) and in MH-S cell line in vitro (flow cytometry). The expression levels of TLR2 and MyD88 in affected lung tissue were also measured to determine their role in the therapy of rSj-Cys on PQ-induced lung injury. RESULT: We identified that treatment with rSj-Cys significantly improved the survival rate of mice with PQ-induced lung injury from 30 % (untreated) to 80 %, reduced the pathological damage of poisoning lung tissue, associated with significantly reduced levels of proinflammatory cytokines (IL-6 from 1490 to 590 pg/ml, TNF-α from 260 to 150 pg/ml) and increased regulatory cytokines (IL-10 from360 to 550 pg/ml, and TGF-ß from 220 to 410 pg/ml) in both sera (proteins) and affected lung tissue (proteins and mRNAs). The polarization of macrophages from M1to M2 type was found to be involved in the therapeutic effect of rSj-Cys on the PQ-induced acute lung injury, possibly through inhibiting TLR2/MyD88 signaling pathway. CONCLUSIONS: Our study demonstrated the therapeutic effect of rSj-Cys on PQ poisoning caused acute lung injury by inducing M2 macrophage polarization through inhibiting TLR2/MyD88 signaling pathway. The finding in this study provides an alternative approach for the treatment of PQ poisoning and other inflammatory diseases.

Effect of the pre-crosslinking of Ba2+ ions on wet spinning of agar fibers.

Decreased coagulation bath concentration and difficult recovery are classical issues observed during the wet spinning of fibers. In this paper, a novel method was presented for preparing environment-friendly agar fibers using deionized water as the coagulation and stretch baths. The addition of Ba2+ into the spinning solution increased the crosslinking time and improved the performance of spinning solution. The results showed that the introduction of Ba2+ in the spinning solution increased the viscosity of the spinning solution. Particularly, when the concentration of BaCl2 in the spinning solution was 7 wt%, the viscosity increased to 39.29 Pa·s, which made the molecular chain arrangement of agar more compact and ordered and promoted the gelation transformation of the spinning solution, resulting in an increase in the gel temperature from 0.2 °C (Ba-0/agar) to 5.4 °C (Ba-7/agar). The spinning solution was more conducive to the formation of fibers in deionized water. In addition, the physical and chemical properties of agar fibers were characterized by X-ray diffraction, Fourier transform infrared spectroscopy, tensile testing, and scanning electron microscopy. The results showed that the use of deionized water as the coagulation bath can improve the color of fiber and solve the problem of fiber adhesion, whereas the mechanical strength of agar fibers with pre-cross-linking metal ions was also improved. For example, the breaking strength of Ba-7/agar/DIW was 0.73 cN/dtex while the breaking strength of Ba-0/agar/DIW was only 0.62 cN/dtex.