CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES ›› 2017, Vol. 35 ›› Issue (2): 110-114.

• ORIGINAL ARTICLES • Previous Articles     Next Articles

Effects of cystatins derived from two species of helminths on the release of nitric oxide and secretion of cytokines from murine peritoneal exudate cells

Xiao-di YANG1, Hui-hui LI1,2, Zhi-yong TAO1, Qiang FANG1, Yang CHENG3, Lan-song XU1, Ren-min XUE1, Yong CHEN1, Hui XIA1, Hui ZHANG1, Hui JIANG1, Tao LIU1, Kun PENG1, Xing-zhi CHEN1,*()   

  1. 1 Department of Microbiology and Parasitology, Anhui Key Laboratory of Infection and Immunity, Bengbu Medical College, Bengbu 233030, China
    2 Department of Histology and Embryology, Bengbu Medical College, Bengbu 233030, China
    3 Department of Patheogenic Infection and Immunity, Wuxi Medical School,Jiangnan University, Wuxi 214122, China
  • Received:2016-12-30 Online:2017-04-20 Published:2017-05-02
  • Contact: Xing-zhi CHEN E-mail:chen007835@163.com
  • Supported by:
    Supported by the Education Commission of Anhui Province(No. KJ2017A235);The Natural Science Fund of Anhui Province（No. 1508085QH158）;The Scientific Research Innovation Team Project of Anhui Colleges and Universities（No. 2016-40）;A Key Supporting Project of Prominent Youth in University of Anhui（No. gxyqZD2016159）;and a National University Students′ Innovation and Entrepreneurship Training Program（No. 201610367009)

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Abstract:

Objective To investigate the effects of cysteine protease inhibitors(cystatins) from Trichinella spiralis (Tscystatin) and Angiostrongylus cantonensis (Accystatin) on the release of nitric oxide (NO) and secretion of cytokines from mouse peritoneal exudate cells (PECs). Methods Ten BALB/c mice were sacrificed under anesthesia to collect intraperitoneal fluid, from which PECs were prepared, consisting mostly of macrophages. The lipopolysaccharide(LPS)-stimulated PECs were incubated with Tscystatin or Accystatin. PECs were assigned into four groups, RPMI group, LPS group, Accystatin + LPS group and Tscystatin + LPS group. The cells in the latter three groups were stimulated with lipopolysaccharides (LPS 2 μg/ml) 2 hours prior to 2 μg/ml Accystatin or Tscystatin treatment for 24 hours. Each group had six replicates. The levels of tumor necrosis factor-α(TNF-α), interleukin IL-6(IL-6) and IL-10 in the culture supernatant were measured by ELISA. The concentration of nitric oxide(NO)was measured by nitrate reductase method. Statistic analysis was performed using SPSS11.5 software. Results The level of pro-inflammatory cytokines TNF-α and IL-6 was(507.50 ± 66.32) and (1 440.49 ± 77.25) pg/ml respectively in the Accystatin + LPS group, significantly higher than (454.15 ± 53.11) and (1 016.27 ± 115.10) pg/ml in LPS group (P < 0.05), but compared with LPS group and Accystatin + LPS group, the level of TNF-α [(296.35 ± 55.30) pg/ml] and IL-6 [(793.54 ± 86.61) pg/ml] in the Tscystatin + LPS group was significantly lower than those in the LPS group and Accystatin + LPS group (P < 0.05). The level of IL-10 was (38.80 ± 6.71) pg/ml in the RPMI group and (53.66 ± 7.72) pg/ml in the LPS group, with no significant difference(P > 0.05). The level of IL-10 in the Accystatin + LPS group [（149.74 ± 26.01) pg/ml] and the Tscystatin + LPS group [(158.76 ± 19.67) pg/ml] was significantly higher than that in the LPS group(P < 0.05). The level of NO in the Accystatin + LPS group and the Tscystatin + LPS group was(12.54 ± 2.12) μmol/L and (7.95 ± 1.40) μmol/L, respectively, significantly decreased compared to the LPS group (P < 0.05). In addition, the concentration NO in the of Tscystatin + LPS group was significantly lower than the Accystatin + LPS group (P < 0.05). Conclusion Cystatins derived from the two species of helminths can both inhibit the release of NO and up-regulate the level of IL-10. Accystatin remarkably promotes the secretion of TNF-α and IL-6, while Tscystatin has the opposite effects.

Key words: Trichinella spiralis, Angiostrongylus cantonensis, Cysteine protease inhibitor

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