Effects of CD38 gene knockout on inflammatory cytokine production in murine spleen B-cells
10.16781/j.0258-879x.2016.11.1373
- Author:
Hui-Yuan JIAO
1
Author Information
1. Department of Microbiology, Faculty of Basic Medical Sciences, Nanchang University
- Publication Type:Journal Article
- Keywords:
B-lymphocytes;
CD38 gene;
Inflammation factors;
Interleukin-1β;
Sirtuins;
Tumor necrosis factor-α
- From:
Academic Journal of Second Military Medical University
2016;37(11):1373-1377
- CountryChina
- Language:Chinese
-
Abstract:
Objective To analyze the number of spleen B-cells and the expression of inflammatory factors and sirtuin 1 (SIRT1) in spleen B-cells of CD38-/- mice, so as to explore the effects of CD38 gene knockout on inflammatory factors in B-cells and its potential mechanism. Methods The DNA levels of CD38 and Neo gene in mouse tail tissues were detected by polymerase chain reaction (PCR). Spleen B-cells from wide-type (WT) C57BL/6 and CD38-/-mice were sorted by magnetic activated cell sorting (MACS), and the purity of sorting B-cells were identified by flow cytometry. The mRNA levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and CD38 gene were detected by real-time PCR, the protein expressions of CD38 and SIRT1 were detected by Western blotting analysis. Results We confirmed the successful establishment of CD38-/-mice and sorted spleen B-cells from WT and CD38-/-mice (purity>95%). Compared with WT mice, the development of spleen was hampered in the CD38-/-mice, the number of spleen cells and spleen B-cells were significantly reduced (P<0. 01), the mRNA levels of TNF-α and IL-1β were significantly decreased (P<0. 01), and the expression level of SIRT1 was significantly increased in CD38-/-mice (P<0. 05). Conclusion CD38 gene knockout can reduce the number of B-cells in the spleen; and it can inhibit the expression of inflammatory factors (TNF-α and IL-1β) in spleen B-cells by activating SIRT1 pathway.
