Background: In the present study, the analgesic and anti-inflammatory effects of alcoholic extract of Elaeagnusangustifolia L.(AEEA) in a model of CIA II-induced osteoarthritis in mice were investigated in vivo and in vitro. Production of inflammatory cytokines such as iL-1β, TNF-α, iL-6, COX-2, iNOS by chondrocytes and synovial macrophages stimulates chondrocytes and produces metalloproteinases, especially MMP-3, MMP-1 and causes destruction of articular cartilage. Mice were divided into three groups: healthy, patient before treatment, and patient after treatment and infected OA using CIA II. After easy death, tryptophan blue cells were cultured using pentobarbital sodium. To determine the injectable doses of the extract, LD50 and LC50 of mouse cells were measured first. It was extracted from the blood of mice and converted to cDNA using a kit. Samples were analyzed by Real-Time PCR to express the expression of iL-1β, TNF-α, iL-6, COX-2, iNOS genes. Chondrocytes were treated with LPS in vitro to produce PGE2, NO. The PGE2 production of chondrocytes was measured by ELISA, and the NO production of chondrocytes was measured by chlorometric method. PGE2 and NO production was performed in three groups: chondrocytes, chondrocytes with LPS, and cells treated with LPS and AEEA.
Results: The results of gene expression in PGE2 were reduced by 89% and NO by 70%. The expression level of COX-2 and iNOS cytokines was significantly decreased after consumption of AEEA extract, and this mice was reported in chondrocytes with 89% decrease for COX-2 gene and 79% decrease for iNOS gene. Expression levels of iL-1β, TNF-α and iL-6 genes decreased by 76% (iL-1β), 89% (TNF-α) and 91% (iL-6), respectively, after consuming AEEA extract. Significant reduction in the expression of pro-inflammatory cytokines by AEEA demonstrates its analgesic and anti-inflammatory effects in mice with osteoarthritis.
Conclusions: AEEA was able to effectively and dose-dependently reduce the production of PGE2, NO in LPS-stimulated chondrocytes (in vitro). It also reduces the expression of pro-inflammatory cytokine genes such as iL-1β, TNF-α, iL-6, COX-2, iNOS in the blood and plasma of mice with osteoarthritis in vivo.
Keywords: Osteoarthritis, CIA II, Elaeagnus angustifolia L., Anti-inflammatory, iL-1β, TNF-α, COX-2, INOS