Combined treatment revealed a great improvement in TIMP-1 levels compared to its levels after using IL-4 treatment alone
Combined treatment revealed a great improvement in TIMP-1 levels compared to its levels after using IL-4 treatment alone. Another important biomarker of RA is IL-1R, a cytokine receptor which binds interleukin-1. and 21. CIA mice were randomly assigned to four groups; group I received an intravenous injection of mouse bone marrow-derived MSCs, while group II received an intraperitoneal injection of IL-4. Group III received a combined treatment of MSC and IL-4, while group IV served as a CIA diseased control group receiving phosphate buffer saline (PBS). A fifth group of healthy mice served as the normal healthy control. To assess changes induced by different treatments, levels of RA-associated inflammatory cytokines and biomarkers were measured in the serum, knee joints, and synovial tissue, using ELISA and Real Time-qPCR. Histopathological features of knee joints were analyzed for all groups. Results showed that combined MSC and IL-4 treatment alleviated signs of synovitis in CIA mice, reverting to the values of healthy controls. This was evident by the decrease in the levels of rheumatic factor (RF), C-reactive protein (CRP) and anti-nuclear antibodies (ANA) by 64, 80, and 71%, respectively, compared to the diseased group. Moreover, tumor MMV008138 necrosis factor-alpha (TNF- ) and monocyte chemoattractant protein-1 (MCP-1) levels decreased by 63 and 68%, respectively. Similarly, our gene expression data showed improvement in mice receiving combined therapy compared to other groups receiving single treatment, where cartilage oligomeric matrix protein (Comp), tissue inhibitor metalloproteinase-1 (Timp1), matrix metalloproteinase1 (Mmp-1), and IL-1 receptor (Il-1r) gene expression levels decreased by 75, 70, and 78%, respectively. Collectively, treatment with a combined therapy of MSC and IL-4 might have an efficient therapeutic effect on arthritis. Thus, further studies are needed to assess the potential of different MSC populations MMV008138 in conjugation with IL-4 in the treatment of experimental arthritis. value 0.05 was considered significant. (MSCs: mesenchymal stem cells, IL-4: interleukin-4, CIA: collagen-induced arthritis, CII: type II collagen) 3.2. Combined MSCs and IL-4 Attenuated RA via Anti-Inflammatory Action After the induction of arthritis in mice, TNF- and MCP-1 KGFR levels increased significantly while IL-10 levels decreased significantly in the synovial tissues of the CIA group, compared to the healthy control group (Figure 3). After using either MSC or IL-4 as a treatment, we found a significant improvement in TNF- synovial levels, by 47% and 48%, compared to the CIA group (Figure 3A). Treatment with the combination of MSCs and IL-4 showed a marked improvement in TNF- levels compared to the diseased levels, by 63% (Figure 3A). MCP-1 levels decreased significantly after treatment with either MSC or IL-4, compared to the CIA group, by 53% and 45%, respectively (Figure 3B). Finally, on using MSCs and IL-4 as a combined treatment, we found a significant improvement in MCP-1 levels, by 68%, compared to the CIA group, with MCP-1 levels almost reaching the same average level as the control group (Figure 3B). The anti-inflammatory cytokine IL-10 levels increased significantly after using MSCs or IL-4 as a treatment, by 62% and 61%, respectively, compared to the CIA group (Figure 3C). After using combined treatment, IL-10 levels increased by 67% compared to the CIA group (Figure 3C). Open in a separate window Figure 3 Decrease in pro-inflammatory and increase in anti-inflammatory cytokine levels in synovial tissue, in CIA mice treated with double therapy of MSCs and IL-4. Comparison of pro-inflammatory (A) TNF- and (B) MCP-1 levels in synovial tissues, in mice treated with CII, compared to the healthy control group. CIA mice treated with MSCs and IL-4, alone or combined, showed attenuation of CIA, where TNF- and MCP-1 decreased following treatment, with a varying degree of significance. Meanwhile, anti-inflammatory (C) IL-10 levels in synovial tissues of CIA mice treated with MSCs and IL-4, alone or combined, showed an increase in levels and attenuation of CIA. Medians were compared using One-way ANOVA, KruskalCWallis test and MMV008138 Dunns multiple comparison test. P value 0.05 was considered significant. 3.3. Combined MSCs and IL-4 Attenuated Articular Cartilage Degradation and Positively Affected Connective Tissue Remodeling of Synovial Tissue Next, we aimed to examine the effect of treatments on joints, along with their cartilage.