Study On The Role Of RLX/RXFP1、MMP9/MMP13and RANKLO/OPG In The Cartilage And Bone Damage Of Arthritis And Chinese Medicine Treatment

PurposeTo study the expression of RLX/RXFP1、MMP-9/MMP-13and RANKL/OPG in theserum and separated PMBCs of CIA rats, and investigate the role of RLX/RXFP1，MMP-9/MMP-13and RANKL/OPG in the pathogenesis of cartilage destruction and boneloss in arthritis as well as the effects of Tripterygium wilfordii polyglycoside on them.MethodsCIA rat model was established and ElISA was applied to detect the protein expression ofRLX, MMP-9, MMP-13, OPG and RANKL in the serum of each group. Real-time PCR wasused to detect the mRNA expression of RXFP1, OPG and RANKL in the PMBCs of eachgroup. The expression of RLX/RXFP1, MMP-9/MMP-13and RANKL/OPG in the modelgroup was compared with the normal group. The expression of RLX/RXFP1,MMP-9/MMP-13and RANKL/OPG after treatment was compared with the model group.Results1. Comparison between the normal group and the CIA model groupSerum RLX, MMP-9, RANKL and RANKL/OPG ratio in the model group weresignificantly higher than that of the normal group (P<0.05), while serum OPG and MMP-13in the model group had no significant difference with the normal group (P>0.05). The mRNAexpression of RXFP1and RANKL in PBMCs of in the model group was significant higherthan that of the normal group (P<0.05), while mRNA expression of OPG in the model grouphad no significant difference with the normal group (P>0.05). 2. Comparison between the TWP group and the model groupSerum RLX, MMP-9, RANKL and RANKL/OPG ratio in the TWP group weresignificantly lower than that of the model group (P<0.05), while serum OPG and MMP-13inthe TWP group had no significant difference with the model group (P>0.05). The mRNAexpression of RXFP1and RANKL in PBMCs of the TWP group was significant lower thanthat of the model group (P<0.05), while mRNA expression of OPG in the TWP group had nosignificant difference with the model group (P>0.05).3. Comparison between the MTX group and the model groupSerum MMP-9, RANKL and RANKL/OPG ratio in the MTX group were significantlylower than that of the model group (P<0.05), while serum RLX, OPG and MMP-13in theMTX group had no significant difference with the model group (P>0.05). The mRNAexpression of RANKL in PBMCs of MTX group was significant lower than that of the modelgroup (P<0.05), while mRNA expression of RXFP1and OPG in the MTX group had nosignificant difference with the model group (P>0.05).4. Comparison between the TWP group and the MTX groupSerum RLX, MMP-9, MMP13, RANKL and RANKL/OPG ratio in the TWP groupwere lower than that of the MTX group without significant difference (P>0.05), while serumOPG in the TWP group was higher than that of the MTX group without significant difference(P>0.05). The mRNA expression of RXFP1and RANKL in PBMCs of TWP group waslower than that of the MTX group (P>0.05), while mRNA expression of OPG in the TWPgroup was higher than that of the MTX group (P>0.05).5. Straight line correlation analysisRLX was positively related to MMP-9and RANKL (r=0.428, P<0.05；r=0.416, P<0.05). Conclusion1. RLX may cause the destruction of the articular cartilage and bone loss byup-regulating the expression of MMP-9/MMP-13and RANKL/OPG;2. Tripterygium wilfordii polyglycoside may reduce the destruction of the articularcartilage and bone loss by inhibiting the RLX/RXFP1-MMP-9-OPG/RANKL. PurposeTo study the expression of RLX/RXFP1、MMP-9/MMP-13and RANKL/OPG in theserum and separated PMBCs of RA patients before and after TWP treatment, and investigateRLX/RXFP1, MMP-9/MMP-13and RANKL/OPG in the pathogenesis of cartilagedestruction and bone loss in RA patients as well as the pharmacological mechanism of TWP.Methods20cases of RA patients with meeting including criteria and30cases of healthyindividuals were included in this study. The blood samples and relative clinical histories ofRA patients before and after TWP treatment as well as healthy individuals were collected.ELISA was applied to detect the expression of RLX-2, MMP-9, MMP-13, OPG and RANKLin the serum. Real-time PCR was used to detect the mRNA expression of RXFP1, OPG andRANKL in the isolated PBMCs. The expression of RLX-2/RXFP1, MMP-9/MMP-13, and OPG/RANKL before and after TWP treatment was compared.ResultsSerum RLX, MMP-9, MMP-13, RANKL and RANKL/OPG ratio as well as the mRNAexpression of RXFP1and RANKL in RA patients had significant difference before and afterTWP treatment (P<0.05or P<0.01). Serum concerntration and mRNA expression of OPG inRA patients had no signicicant difference before and after TWP treatment (P>0.05).Conclusion1. RLX may up-regulate the expression of MMP-9and RANKL to induce cartilagedestruction and bone loss involved in the mechanism of arthritis;2. The TWP may ease the cartilage destruction and bone loss of RA by inhibiting RLX/RXFP1-MMP-9-OPG/RANKL. PurposeTo study the expression of RLX/RXFP1、MMP-9/MMP-13and RANKL/OPG in theserum and separated PMBCs of RA and OA patients with different syndromes, andinvestigate the role of RLX/RXFP1, MMP-9/MMP-13and RANKL/OPG in the pathogenesisof cartilage destruction in arthritis.Methods Clinical histories and blood samples of94untreated patients with arthritis (including56cases of RA patients and38cases of OA patients) as well as30cases of healthy individualswere collected. ELISA was applied to detect the expression of RLX-2, MMP-9, MMP-13,OPG and RANKL in serum. Real-time PCR was used to detect the mRNA expression ofRXFP1, OPG and RANKL in the isolated PBMCs. The expression of RLX/RXFP1,MMP-9/MMP-13, and OPG/RANKL among RA group, OA group and normal groups wascompared. Multiple linear regression analysis was applied to analyze the correlation of serumRLX-2, MMP-9, MMP-13, OPG and RANKL with clinical indicators in RA and OA patients.Straight line correlation analysis was used to analyze the correlation of serum RLX-2withMMP-9and RANKL.ResultsSerum RLX-2and the mRNA expression of RXFP1in isolated PBMCs were increasedin RA and OA patients compared to the normal group (P<0.05or P<0.01), and there was nosignificant difference between RA and OA patients (P>0.05). Serum MMP-9was increased inRA and OA patients compared to the normal group (P<0.05or P<0.01), and there wassignificant difference between RA and OA patients (P<0.05). Serum MMP-13was lower inRA patients than that of OA patients but higher than that of the normal group (P>0.05).Serum OPG as well as OPG mRNA expression in isolated PBMCs were lower in RA patientsthan that of OA patients and normal group (P>0.05). Serum RANKL as well as RANKLmRNA expression in isolated PBMCs increased in the RA patients compared to OA patientsand the normal group (P<0.05or P<0.01). Serum RANKL/OPG ratio was increased in RAand OA patients compared to the normal group (P<0.05or P<0.01), and there was significantdifference between RA and OA patients (P<0.05). Serum MMP-9and RANKL as well asRANKL mRNA expression in isolated PBMCs of RA patients differentiated as Cold or Heat Syndrome were higher than that of OA patients differentiated as Cold or Heat Syndromerespectively(P<0.05).Multiple linear regression analysis showed that serum levels of RLX-2and MMP-9were positively correlated with ESR(β=0.417, t=2.126, P=0.039；β=0.435, t=2.328, P=0.042).Straight line correlation analysis showed RLX-2was significantly positively related toMMP-9and RANKL (r=0.417, P<0.01; r=0.423, P<0.01).Conclusion1. RLX-2may induce cartilage destruction and bone loss in RA and OA byup-regulating MMP-9/MMP-13and RANKL/OPG;2. Both RLX-2and MMP-9reflected the inflammation level of RA.