Regulatory Effects Of P53 In Synoviocytes On Th17, Treg, Th1 And Th2 From Rheumatoid Arthritis

Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by chronic destructive arthritis with unknown cause, involving small joints such as proximal interphalangeal joints, wrists, ankles and knees. Pathologically, chronic inflammation and hyperplasia of synovial membrane forms pannus, and invades cartilage, subcartilage, ligament and texture, resulting in the destruction of bone, cartilage and articular capsule, leading to malformation and dysfunction of those joints.In our previous work, we had mastered the skills of culturing primary human fibroblast-like synoviocytes (FLS), identified the appropriate method to interfere the expression of p53 in FLS by siRNA technique. In this study, we cocultured FLS transfected with p53 siRNA with CD4~+ T lymphocytes from RA patients, looking for the corresponding changes in FLS and the effects of p53 suppression in FLS on Th17, Treg, Th1 and Th2 from RA patients. Our work provided a new way to investigate the cause and pathophysiological changes of RA.This paper included two parts:1. Regulatory effects of p53 in synoviocytes on Th17, Treg, Th1 and Th2 from peripheral blood of RA patientsOn the 3rd day after the p53 in FLS was interfered by siRNA technology, CD4~+ T lymphocytes from RA was prepared by magnetic technology. After cocultured for 48h, the compositions of Th17, Treg, Th1, Th2 cells were examined by flow cytometry (FCM). At the same time, Reverse transcription-Polymerase chain reaction (RT-PCR) was conducted to investigate the expressions of IFNγ,RORγt,1L17,Foxp3 in mRNA levels. The results showed that after p53 in FLS was interfered, the intracellular levels of IFNγand IL17 increased in CD4~+ T lymphocytes, while no significant changes were found in Th2 and Treg.2. The biological changes of FLS after p53 was interfered.On the 5th day after the p53 in FLS was interfered by siRNA technology, FLS were collected. RT-PCR was used to investigate the differential changes of osteoprotegerin (OPG) in mRNA levels, and Enzyme-linked immunosorbent assay (ELISA) was used to test IL6 level in culture media. The results showed that after p53 in FLS was interfered, the expression of OPG did not change obviously, while IL6 in culture media decreased.To date, our result is the first time to investigate the regulatory effects of p53 in human synoviocytes on Th17, Treg, Th1 and Th2 from RA patients, and on OPG and IL6 synthesis in p53-interfered FLS per se.