ORIGINAL ARTICLE

Naunyn-Schmiedeberg's Archives of Pharmacology

, 381:83

First online:

Protosappanin A induces immunosuppression of rats heart transplantation targeting T cells in grafts via NF-κB pathway

  • Jian WuAffiliated withDepartment of Cardiology, The Second Affiliated Hospital of Harbin Medical University
  • , Maomao ZhangAffiliated withDepartment of Cardiology, The Second Affiliated Hospital of Harbin Medical University
  • , Haibo JiaAffiliated withDepartment of Cardiology, The Second Affiliated Hospital of Harbin Medical University
  • , Xingtao HuangAffiliated withDepartment of Cardiology, The Second Affiliated Hospital of Harbin Medical University
  • , Qi ZhangAffiliated withDepartment of Cardiology, The Second Affiliated Hospital of Harbin Medical University
  • , Jingbo HouAffiliated withDepartment of Cardiology, The Second Affiliated Hospital of Harbin Medical UniversityThe Cardiovascular Department, The Second Affiliated Hospital of Harbin Medical University Email author 
  • , Yu BoAffiliated withDepartment of Cardiology, The Second Affiliated Hospital of Harbin Medical University

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Abstract

Protosappanin A as one major and effective ingredient from Caesalpinia sappan L. exhibited antirejection activity obviously in heart-transplanted rat. The present study was designed to screen out the potential target genes of protosappanin A with microarray technology and reveal some molecular mechanism of immunosuppressive effect. Rats performed with ectopic peritoneal heart transplantation were randomized into three groups receiving different treatments for 7 days: protosappanin A group (25 mg kg−1), cyclosporine A group (10 mg kg−1), and control group. The differentially expressed genes responding to protosappanin A were analyzed with microarrays. Among common differentially expressed genes, the ones of interest were selected for further evaluation by real-time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), Western blot, immunochemistry, immunofluorescence, and ELISA. Among the 146 common differentially expressed genes, NF-κB and related genes like IκBa, IFN-r, and IP10 were selected for verification. The results of qRT-PCR, Western blot, immunochemistry, and ELISA showed that protosappanin A significantly reduced the expression of NF-κB, IFN-r, and IP10 (p < 0.05) and increased IκBa expression (p < 0.05) in graft. Moreover, the immunochemistry staining of NF-κB and IκBa was mainly observed in infiltrating mononuclear cells. Strikingly, immunofluorescent staining localized NF-κB to the TCR-positive T cells in graft. Furthermore, protosappanin A exhibited inhibitory effect on T cell proliferation in recipients after 7-day treatment. In conclusion, protosappanin A might act on T cells through inhibiting NF-κB activation and downstream gene expressions of IFN-r and IP10, meanwhile reducing T cell proliferation responding to alloantigen, so as to induce immunosuppressive effect. The results encourage a potential therapeutic evaluation of protosappanin A for clinical organ transplantation or other T cell-mediated immune disorders. Additionally, our study also verified the feasibility of microarray utilization in Chinese herb research to explore molecular mechanism and promote development of scientific theories.

Keywords

Protosappanin A Heart transplantation Immunosuppression Microarray Nuclear factor kappa B T cells