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Expression of proteinase-activated receptor (PAR)-2 in monocytes from allergic patients and potential molecular mechanism

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Abstract

Serine proteases play an important role in inflammation via PARs. However, little is known of expression levels of PARs on monocytes of allergic patients, and influence of serine proteases and PARs on TNF-α secretion from monocytes. Using quantitative real-time PCR (qPCR) and flowcytometry techniques, we observed that the expression level of PAR-2 in monocytes of patients with allergic rhinitis and asthma was increased by 42.9 and 38.2 %. It was found that trypsin, thrombin, and tryptase induced up to 200, 320, and 310 % increase in TNF-α release from monocytes at 16 h, respectively. PAR-1 agonist peptide, SFLLR-NH2, and PAR-2 agonist peptide tc-LIGRLO-NH2 provoked up to 210 and 240 % increase in release of TNF-α. Since SCH 79797, a PAR-1 antagonist, and PD98059, an inhibitor of ERK inhibited thrombin- and SFLLR-NH2-induced TNF-α release, the action of thrombin is most likely through a PAR-1- and ERK-mediated signaling mechanism. Similarly, because FSLLRN-NH2, an inhibitor of PAR-2 diminished tryptase- and tc-LIGRLO-NH2-induced TNF-α release, the action of tryptase appears PAR-2 dependent. Moreover, in vivo study showed that both recombinant cockroach major allergens Per a 1 and Per a 7 provoked upregulation of PAR-2 and PAR-1 expression on CD14+ cells in OVA-sensitized mouse peritoneum. In conclusion, increased expression of PAR-2 in monocytes of AR and asthma implicates that PAR-2 likely play a role in allergy. PAR-2- and PAR-1-mediated TNF-α release from monocytes suggests that these unique protease receptors are involved in the pathogenesis of inflammation.

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Abbreviations

PARs:

Proteinase-activated receptors

MMP:

Matrix metalloprotease

SBTI:

Soybean trypsin inhibitor

OVA:

Ovalbumin

BSA:

Bovine serum albumin

FCS:

Fetal calf serum

AR:

Allergic rhinitis

HC:

Healthy control subjects

ARIA:

Allergic Rhinitis and its Impact on Asthma

PBMC:

Peripheral blood mononucleated cells

MACS:

Magnet-activated cell sorting

CASE:

Cellular activation of signaling ELISA

ERK:

Extracellular signal-regulated kinase

TGF:

Transforming growth factor

TNF:

Tumor necrosis factor

VCAM:

Vascular cell adhesion molecule

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Acknowledgments

This project was sponsored by the grants from the “12th Five-Year ” National Science and Technology Support Plan (2014BAI07B02), the National Natural Science Foundation of China (nos. 81172836, 81471592, 81472016 ); Major Science and Technology Platform for Institution of Higher Education in Liaoning Province (2014168); “Twelfth five-year” Public Welfare Industry Special Scientific Research Project (2015SQ00136); the National Natural Science Foundation of Liaoning Province (2014022027, 2014022019); Program for Liaoning Innovation Research Team in University (LNIRT, LT2013017); Climbing Scholar Project for Institution of Higher Education in Liaoning province (2013222); Allergic Disease Translational Medicine Research Centre of Liaoning Province (201341); Liaoning Provincial Engineering Research Centre for Diagnosing & Treating Inflammatory Disease (20141093); Clinical Capability Construction Project for Liaoning Provincial Hospitals ( LNCCC-A06-2014); and Science and Technology Planning Project of.

Suzhou (SYS201272).

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Authors and Affiliations

  1. Allergy and Clinical Immunology Research Centre, the First Affiliated Hospital of Liaoning Medical University, No. 2, Section 5, Renmin Street, Guta District, Jinzhou, Liaoning, 121001, People’s Republic of China

    Shuqing Ge, Zhang Huiyun & Shaoheng He

  2. Department of Dentistry, the First Affiliated Hospital of Liaoning Medical University, Jinzhou, Liaoning, 121001, China

    Shuqing Ge & Bin Zhang

  3. Department of Infectious Diseases, Shantou University Medical College, Shantou, 515031, China

    Tao Li, Qijian Yao & Yanshan Zheng

  4. Clinical Research Centre, the First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, 210029, China

    Hongling Yan

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  1. Shuqing Ge
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Correspondence to Shaoheng He.

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The authors declare that there is no competing interest regarding the publication of this article.

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Shuqing Ge and Tao Li contributed equally to the study

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Ge, S., Li, T., Yao, Q. et al. Expression of proteinase-activated receptor (PAR)-2 in monocytes from allergic patients and potential molecular mechanism. Cell Biol Toxicol 32, 529–542 (2016). https://doi.org/10.1007/s10565-016-9353-x

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  • DOI: https://doi.org/10.1007/s10565-016-9353-x

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