| Abstrakt: |
Objective: To observe the anti-inflammatory and glycometabolic effects of glutathione (GSH) on macrophages in collagen induced arthritis (CIA) mice. Methods: 1 CIA model establishment and groups: A total of 14 female DBA/1J mice were randomly divided into: CIA+PBS group and CIA+GSH group. The mice were sacrificed on the 50th day, collecting serum and isolating bone marrow derived macrophages (BMDM), which were marked as BMDM1. 2 Trained immunity model establishment and groups: BMDM were isolated from normal DBA/1J mice, and were pretreated with histone H3K27 demethylases inhibitor (GSKJ1) or PBS for 2 h. Then, serum from CIA model mice in vivo was incubated for 24 h, and the samples were grouped as follows: (CIA+GSH)+PBS group, (CIA+GSH)+GSKJ1 group, (CIA+PBS)+PBS group, (CIA+PBS)+GSKJ1 group. Lipopolysaccharide (LPS) was adopted to stimulated cells on the 6th day, which were marked as BMDM2. 3 RNA sequencing was used to detect differentially expressed genes (DEGs) and their function in BMDM1 and BMDM2. q-PCR was adopted to estimate the mRNA levels of PFK and Idh3g. The culture supernatants were used to measure the protein levels of TNF-α and IL-6 by ELISA. Results: 1 Compared with CIA+PBS group, the mice in CIA+GSH group showed lighter of joint swelling (P<0.05), less arthritis index (P<0.05), HE staining suggested less inflammatory cell infiltration, Safranin O-fast green staining showed more chondrocytes, TRAP staining indicated reduced osteoclasts. 2 In BMDM1, GO analysis showed that DEGs were mainly involved in glutathione derivative metabolic process, IL-6 production, inflammatory response, innate immune response, regulation of primary metabolic process and glycolipid binding, compared with CIA+ GSH and CIA+PBS group. In CIA+GSH group, the mRNA level of PFK was significantly decreased (P<0.05), while Idh3g was also significantly up-regulated (P<0.05), and the expression of TNF-α, IL-6 were both reduced (P<0.05) compared with CIA+PBS group. 3 In BMDM2, GO analysis showed that DEGs were also involved in inflammatory response, activation of innate immune response, regulation of tumor necrosis factor production, positive regulation of IL-6 production, regulation of glycolytic process and 1, 3- β-Dglucan binding between CIA+GSH and CIA+PBS group. Furthermore, in (CIA+GSH)+PBS group, PFK was decreased (P<0.05), Idh3g was up-regulated (P<0.05), and IL-6 was also significantly down-regulated (P<0.05) compared with (CIA+PBS)+PBS group. However, there was no significant difference in the expression of Idh3g and PFK, moreover, TNF-α and IL-6 were significantly upregulated compared with (CIA+GSH)+GSKJ1 group and (CIA+GSH)+PBS group. Conclusion: GSH can regulate glycometabolism and inflammatory response of macrophages via demethylation of histone H3K27, and it can also alleviate CIA in mice. [ABSTRACT FROM AUTHOR] |
