Transplantation of IL‑1β siRNA‑modified bone marrow mesenchymal stem cells ameliorates type II collagen‑induced rheumatoid arthritis in rats

Abstract

Rheumatoid arthritis (RA) is a chronic autoimmune disease that causes erosion of articular cartilage and bone and has adverse effects on both patients and livestock animals. The aim of the present study was to investigate the role of interleukin-1β (IL-1β) in the pathogenesis of RA, and to further determine whether injection of IL-1β small interfering RNA (siRNA) or transplantation of IL-1β siRNA + bone marrow mesenchymal stem cells (BMSCs) can ameliorate RA in rats. A collagen-induced arthritis (CIA) rat model was established by injecting type II collagen for 4 weeks. Next, CIA rats were randomly divided into three groups and injected or transplanted with PBS, IL-1β siRNA and IL-1β siRNA + BMSCs for another 4 weeks. The CIA rat model was successfully established, as demonstrated by the higher toe swelling value, thymus and spleen/body weight, immobility time and serum IL-1β concentration, as well as lower body weight, climbing time and mRNA expression of programmed death-1 (PD-1), transforming growth factor-β1 (TGF-β1) and forkhead box protein 3 (Foxp3) in the spleen, compared with control rats. Furthermore, histopathology results demonstrated that joint swelling and redness were observed in the knee joints of CIA rats. H&E results revealed that CIA rats presented erosive destruction of the bone and ulceration of the articular cartilage. In addition, in vitro results demonstrated that IL-1β expression was successfully silenced after IL-1β siRNA transfection in lipopolysaccharide-stimulated BMSCs. When compared with the results of PBS rats, both IL-1β siRNA injection and IL-1β siRNA + BMSC transplantation significantly increased the body weight, climbing time and mRNA expression of PD-1, TGF-β1 and Foxp3 in the spleen, while significantly reduced the immobility time and serum IL-1β concentration. In addition, when compared with that of IL-1β siRNA injection, IL-1β siRNA + BMSC transplantation exhibited markedly higher therapeutic efficacy against CIA. These results demonstrated that higher IL-1β contributed to the pathogenesis of CIA, and that IL-1β siRNA injection ameliorated CIA, while its combination with BMSCs exerted synergistic effects, which may be beneficial against RA.

Effect and Mechanism of siRNAs Targeting IL-1β/TNF-α Combined with BMSCs Transplantation in Ameliorating Rheumatoid Arthritis in Rats

“…In vitro results demonstrated that transfection with IL-1β/TNF-α siRNAs significantly reduced the contents of IL-1β and TNF-α in both cell supernatant and cell lysate in LPS-stimulated RAW264.7 compared with the control cells. These results indicated that exogenous IL-1β/TNF-α siRNAs were able to be transfected into RAW264.7 successfully and produces a highly stable silencing effect, which was consistent with our previous result [ 12 ]. Moreover, the IL-1β and TNF-α contents in the serum and the mRNA expression of IL-1β and TNF-α in spleen in IL-1β/TNF-α siRNAs rats were obviously decreased relative to PBS rats.…”

“…On the other hand, BMSCs alleviate the autoimmune response by regulating the balance between Th17/Treg [ 39 ]. BMSCs have also been shown to be effective in the treatment of osteoarthritis [ 40 , 41 ], degenerative arthritis [ 42 , 43 ] and RA in animals [ 12 ]. In particular, BMSCs has a positive effect on the articular cartilage surface repair [ 40 ].…”

“…Eight-week-old female Wistar rats (200 ± 5 g) were gained from the Comparative Medicine Cente, Yangzhou University (certificate of quality is SYXK (Su) 2017-0007). The CIA model was induced as described previously [ 12 ]. Briefly, thirty-five rats were injected with chicken type Ⅱ collagen (cat.…”

“…BMSCs were derived from 6-week-old Wistar rats as previously described [ 12 ]. Anesthesia was performed by intraperitoneal injection of pentobarbital sodium (65 mg/kg body weight, Sigma-Aldrich, Merck KGaA.…”

“…In addition, BMSCs have a regulatory effect on immunosuppression through prostaglandin metabolites and macrophage clearance [ 11 ]. Our previous study preliminarily showed that BMSCs had good therapeutic effects on RA rats [ 12 ]. However, the effect of siRNAs targeting IL-1β/TNF-α combined with BMSCs transplantation in ameliorating RA in rats and its potential mechanism are largely unknown.…”