Limits...
(5R)-5-Hydroxytriptolide (LLDT-8) inhibits osteoclastogenesis via RANKL/RANK/OPG signaling pathway.

Shen Y, Jiang T, Wang R, He S, Guo M, Zuo J, He D - BMC Complement Altern Med (2015)

Bottom Line: In addition, LLDT-8 decreased the number of TRAP-positive cells derived from RAW264.7 in the presence of RANKL and M-CSF.Furthermore, LLDT-8 also inhibited the expression of p-IκB, a key regulator of RANKL signaling pathway.LLDT-8 exerts its anti-osteoclastogenesis effect in RA probably through regulating RANKL/RANK/OPG system and its downstream signaling pathway as well as cytokine productions.

View Article: PubMed Central - PubMed

Affiliation: Department of Rheumatology, Shanghai Guanghua Hospital of Integrated Traditional and Western Medicine, Shanghai, 200052, China. shenyi_740@hotmail.com.

ABSTRACT

Background: The aim of this study was to investigate the regulative activity of (5R)-5-hydroxytriptolide (LLDT-8) on receptor activator of nuclear factor κ-B ligand (RANKL)/receptor activator of nuclear factor κ-B (RANK)/Osteoprotegerin (OPG) system in rheumatoid arthritis (RA) and its anti-osteoclastogenesis mechanism.

Methods: The expression of OPG, RANK and RANKL in CD3(+) T leukomonocytes in both peripheral blood and synovial fluid of RA patients was evaluated by flow cytometry. The levels of interleukin (IL) 1β, IL-6, IL-10, IL-21 and IL-23 in the supernatants of peripheral blood mononuclear cells (PBMCs) and synovial fluid mononuclear cells (SFMCs) were assayed by ELISA. Tartaric acid phosphatase (TRAP) staining was used to identify the osteoclast-like cells derived from RAW264.7. Western blotting analysis was used to check the downstream molecules of RANKL.

Results: LLDT-8 increased the rate of OPG expression in CD3(+) T leukomonocytes in peripheral blood as well as the ratio of OPG/RANKL in both peripheral blood and synovial fluid. LLDT-8 inhibited IL-1β, IL-6, IL-21 and IL-23 secretion, but promoted the secretion of IL-10 in the supernatants of PBMCs and SFMCs. In addition, LLDT-8 decreased the number of TRAP-positive cells derived from RAW264.7 in the presence of RANKL and M-CSF. Furthermore, LLDT-8 also inhibited the expression of p-IκB, a key regulator of RANKL signaling pathway.

Conclusions: LLDT-8 exerts its anti-osteoclastogenesis effect in RA probably through regulating RANKL/RANK/OPG system and its downstream signaling pathway as well as cytokine productions.

Show MeSH

Related in: MedlinePlus

LLDT-8 increased the ratio of OPG/RANKL in synovial fluid of RA patients. SFMCs were isolated from synovial fluid of RA patients and treated with various concentrations of LLDT-8 (0, 25 and 50 nM, respectively) for 24 hours, then stained and detected by flow cytometry. (A) The psuedocolor plots of OPG, RANK and RANKL. (B) The rate of OPG expression on CD3+ T leukomonocyte in the three groups. (C) The ratio of OPG/RANKL on CD3+ T cells in the three groups. *p < 0.05 compared to the control group (0 nM).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4379732&req=5

Fig2: LLDT-8 increased the ratio of OPG/RANKL in synovial fluid of RA patients. SFMCs were isolated from synovial fluid of RA patients and treated with various concentrations of LLDT-8 (0, 25 and 50 nM, respectively) for 24 hours, then stained and detected by flow cytometry. (A) The psuedocolor plots of OPG, RANK and RANKL. (B) The rate of OPG expression on CD3+ T leukomonocyte in the three groups. (C) The ratio of OPG/RANKL on CD3+ T cells in the three groups. *p < 0.05 compared to the control group (0 nM).

Mentions: We also isolated SFMCs from synovial fluid of RA patients, and examined the OPG, RANK and RANKL expression on these cells treated by the same method as PBMCs. The results showed LLDT-8 (50 nM) increased the ratio of OPG/RANKL on CD3+ T cells in synovial fluid (Figure 2C), and there was an upward trend on OPG expression (Figure 2B). However, there was no distinct effect of LLDT-8 on the rate of RANKL or RANK expression on CD3+ T cells in synovial fluid.Figure 2


(5R)-5-Hydroxytriptolide (LLDT-8) inhibits osteoclastogenesis via RANKL/RANK/OPG signaling pathway.

Shen Y, Jiang T, Wang R, He S, Guo M, Zuo J, He D - BMC Complement Altern Med (2015)

LLDT-8 increased the ratio of OPG/RANKL in synovial fluid of RA patients. SFMCs were isolated from synovial fluid of RA patients and treated with various concentrations of LLDT-8 (0, 25 and 50 nM, respectively) for 24 hours, then stained and detected by flow cytometry. (A) The psuedocolor plots of OPG, RANK and RANKL. (B) The rate of OPG expression on CD3+ T leukomonocyte in the three groups. (C) The ratio of OPG/RANKL on CD3+ T cells in the three groups. *p < 0.05 compared to the control group (0 nM).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4379732&req=5

Fig2: LLDT-8 increased the ratio of OPG/RANKL in synovial fluid of RA patients. SFMCs were isolated from synovial fluid of RA patients and treated with various concentrations of LLDT-8 (0, 25 and 50 nM, respectively) for 24 hours, then stained and detected by flow cytometry. (A) The psuedocolor plots of OPG, RANK and RANKL. (B) The rate of OPG expression on CD3+ T leukomonocyte in the three groups. (C) The ratio of OPG/RANKL on CD3+ T cells in the three groups. *p < 0.05 compared to the control group (0 nM).
Mentions: We also isolated SFMCs from synovial fluid of RA patients, and examined the OPG, RANK and RANKL expression on these cells treated by the same method as PBMCs. The results showed LLDT-8 (50 nM) increased the ratio of OPG/RANKL on CD3+ T cells in synovial fluid (Figure 2C), and there was an upward trend on OPG expression (Figure 2B). However, there was no distinct effect of LLDT-8 on the rate of RANKL or RANK expression on CD3+ T cells in synovial fluid.Figure 2

Bottom Line: In addition, LLDT-8 decreased the number of TRAP-positive cells derived from RAW264.7 in the presence of RANKL and M-CSF.Furthermore, LLDT-8 also inhibited the expression of p-IκB, a key regulator of RANKL signaling pathway.LLDT-8 exerts its anti-osteoclastogenesis effect in RA probably through regulating RANKL/RANK/OPG system and its downstream signaling pathway as well as cytokine productions.

View Article: PubMed Central - PubMed

Affiliation: Department of Rheumatology, Shanghai Guanghua Hospital of Integrated Traditional and Western Medicine, Shanghai, 200052, China. shenyi_740@hotmail.com.

ABSTRACT

Background: The aim of this study was to investigate the regulative activity of (5R)-5-hydroxytriptolide (LLDT-8) on receptor activator of nuclear factor κ-B ligand (RANKL)/receptor activator of nuclear factor κ-B (RANK)/Osteoprotegerin (OPG) system in rheumatoid arthritis (RA) and its anti-osteoclastogenesis mechanism.

Methods: The expression of OPG, RANK and RANKL in CD3(+) T leukomonocytes in both peripheral blood and synovial fluid of RA patients was evaluated by flow cytometry. The levels of interleukin (IL) 1β, IL-6, IL-10, IL-21 and IL-23 in the supernatants of peripheral blood mononuclear cells (PBMCs) and synovial fluid mononuclear cells (SFMCs) were assayed by ELISA. Tartaric acid phosphatase (TRAP) staining was used to identify the osteoclast-like cells derived from RAW264.7. Western blotting analysis was used to check the downstream molecules of RANKL.

Results: LLDT-8 increased the rate of OPG expression in CD3(+) T leukomonocytes in peripheral blood as well as the ratio of OPG/RANKL in both peripheral blood and synovial fluid. LLDT-8 inhibited IL-1β, IL-6, IL-21 and IL-23 secretion, but promoted the secretion of IL-10 in the supernatants of PBMCs and SFMCs. In addition, LLDT-8 decreased the number of TRAP-positive cells derived from RAW264.7 in the presence of RANKL and M-CSF. Furthermore, LLDT-8 also inhibited the expression of p-IκB, a key regulator of RANKL signaling pathway.

Conclusions: LLDT-8 exerts its anti-osteoclastogenesis effect in RA probably through regulating RANKL/RANK/OPG system and its downstream signaling pathway as well as cytokine productions.

Show MeSH
Related in: MedlinePlus