Objective:To explore immunoregulatory function of long non-coding RNA Gm26688 on macrophages RAW264.7,macrophages stimulated by LPS were treated with Chinese giant salamander(CGS)collagen peptides to explore role of Gm26688 in mechanism of CGS collagen peptides in regulating inflammatory reflection.Methods:Gm26688 sequence was cloned by RT-PCR,Gm26688 localization in macrophage was analyzed by RNA-FISH;qRT-PCR was performed to detect expressions of Gm26688 and inflammatory factors after LPS and CGS bone collagen peptides treatment;expression of Gm26688 was silenced by small interfering RNA(siRNA),and qRT-PCR was used to detect effects of LPS and CGS bone collagen peptides on expression of inflammatory factors after Gm26688 interference;Western blot was used to detect protein expression of NF-κB p65 and its phosphorylation level.Results:LPS significantly promoted expression of Gm26688.RT-PCR amplified Gm26688 sequence,and RNA-FISH showed that Gm26688 was mainly expressed in nucleus.CGS bone collagen peptides inhibited expression of Gm26688 induced by LPS,and<3 kD CGS bone collagen peptides had the most significant inhibitory effect on Gm26688.CGS bone collagen peptides(<3 kD)significantly inhibited LPS-induced expressions of inflammatory factors IL-6,TNF-α and IL-1β.Interference of Gm26688 significantly down-regulated expressions of inflammatory factors IL-6 and TNF-α in LPS or LPS+CGS bone collagen peptides-stimulated RAW264.7 cells.Western blot results showed that CGS bone collagen peptides down-regulated level of NF-κB p65 phosphorylation through Gm26688.Conclu-sion:CGS bone collagen peptides inhibit LPS-induced inflammation in RAW264.7 cells via Gm26688/NF-κB pathway.

Objective:To explore immunoregulatory function of long non-coding RNA Gm26688 on macrophages RAW264.7,macrophages stimulated by LPS were treated with Chinese giant salamander(CGS)collagen peptides to explore role of Gm26688 in mechanism of CGS collagen peptides in regulating inflammatory reflection.Methods:Gm26688 sequence was cloned by RT-PCR,Gm26688 localization in macrophage was analyzed by RNA-FISH;qRT-PCR was performed to detect expressions of Gm26688 and inflammatory factors after LPS and CGS bone collagen peptides treatment;expression of Gm26688 was silenced by small interfering RNA(siRNA),and qRT-PCR was used to detect effects of LPS and CGS bone collagen peptides on expression of inflammatory factors after Gm26688 interference;Western blot was used to detect protein expression of NF-κB p65 and its phosphorylation level.Results:LPS significantly promoted expression of Gm26688.RT-PCR amplified Gm26688 sequence,and RNA-FISH showed that Gm26688 was mainly expressed in nucleus.CGS bone collagen peptides inhibited expression of Gm26688 induced by LPS,and<3 kD CGS bone collagen peptides had the most significant inhibitory effect on Gm26688.CGS bone collagen peptides(<3 kD)significantly inhibited LPS-induced expressions of inflammatory factors IL-6,TNF-α and IL-1β.Interference of Gm26688 significantly down-regulated expressions of inflammatory factors IL-6 and TNF-α in LPS or LPS+CGS bone collagen peptides-stimulated RAW264.7 cells.Western blot results showed that CGS bone collagen peptides down-regulated level of NF-κB p65 phosphorylation through Gm26688.Conclu-sion:CGS bone collagen peptides inhibit LPS-induced inflammation in RAW264.7 cells via Gm26688/NF-κB pathway.