The Effects of Pricking Blood Therapy on Promotor Methylation of IL-1β， IL-10 in Rats Local Ankle Joint with Acute Gouty Arthritis Model

OBJECTIVE To observe the cross section diameter and the effects of pricking blood therapy on acute gouty arthrits rats model's interleukin-1β (IL-1β)， interleukin -10 (IL-10) mRNA expression and promoter methylation in local ankle， and explore the epigenetics mechanisms of pricking blood therapy's treatment. METHODS 36 SD rats were randomly divided into normal control group， urate arthritis group， ibuprofen group and bloodletting group. The acute gouty arthrits model set up by injecting uric acid into the right ankle joint cavity .The ibuprofen group was treated by gastric perfusion of ibuprofen， while the bloodletting group was pricked at the right Kunlun points. The swelling index of the modeling joint was evaluated， and the morphological changes were observed under light microscope. The mRNA expression levels and promoter methylation status of IL-1β and IL-10 in rats local ankle were detected by qPCR and pyrosequencing detection. RESULTS Compared with the urate arthritis group and ibuprofen group， the joint swelling index in the bloodletting group was decreased (P<0.01) after the treatment. Pricking blood therapy reduced intracellular deposition of uric acid in the joint cavity， and inhibited inflammatory cells to improve the synovial membrane tissue. Compared with urate arthritis group， the IL-1β mRNA expression levels of blood group was decreased significantly (P<0.01). IL-1β promoter methylation and mRNA expression didn't showed a negative correlation. Compared with normal control group， urate arthritis group and ibuprofen group， the expression levels of IL-10 mRNA in blood group increased significantly (P<0.01)， and the average methylation rate of IL-10 among blood group decreased significantly (P<0.05~0.01). IL-10 promoter methylation and mRNA expression showed a strong negative correlation (r=-0.899， P<0.01). CONCLUSION In epigenetics， the methylation of IL-10 can regulate the gene expression， which can be one of important mechanisms which shows anti-inflammatory effects of pricking blood therapy， .