RLX/RXFP1、MMP9/MMP13和RANKL/OPG在关节炎软骨和骨破坏及中药治疗中的作用
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摘要
目的
通过建立CIA大鼠模型，研究RLX/RXFP1、MMP-9/MMP-13和RANKL/OPG在CIA大鼠模型血清和外周单核细胞中的表达水平，以及探讨RLX/RXFP1、MMP-9/MMP-13和RANKL/OPG在关节炎软骨破坏和骨质丢失中的机制和中药治疗中的作用。
方法
建立CIA大鼠模型，应用ELISA方法检测各组大鼠血清中RLX、MMP-9、MMP-13、OPG和RANKL的含量；采用荧光实时定量PCR方法检测各组大鼠的外周单核细胞中RXFP1、OPG和RANKL的mRNA的表达。比较CIA模型组和正常组之间RLX/RXFP1、MMP-9/MMP-13和RANKL/OPG的表达有无差异；比较药物治疗后与模型组之间RLX/RXFP1、MMP-9/MMP-13和RANKL/OPG的表达有无差异。
结果
1.正常组与CIA模型组比较
CIA模型组中大鼠血清RLX、MMP-9、RANKL的含量以及RANKL/OPG比值较正常组明显增高，差异有统计学意义(P<0.05)；而OPG和MMP-13的含量与正常组无统计学差异(P>0.05)；CIA模型组中大鼠的外周单核细胞中RXFP1和RANKL的mRNA水平的表达较正常组中增高，具有统计学差异(P<0.05)，而OPG的表达与正常组无统计学差异(P>0.05)。
2.雷公藤多甙组与模型组比较
雷公藤多甙组中大鼠血清中RLX、MMP-9、RANKL的含量以及RANKL/OPG比值较模型组明显降低，差异有统计学意义(P<0.05)；而OPG和MMP-13的含量与模型组无统计学差异(P>0.05)；雷公藤多甙组大鼠外周单核细胞中RXFP1和RANKL的mRNA水平的表达较模型组中降低，具有统计学差异(P<0.05)，而OPG的表达与模型组无统计学差异(P>0.05)。
3.甲氨喋呤组与模型组比较
甲氨喋呤组中大鼠血清中MMP-9和RANKL的含量以及RANKL/OPG比值较模型组明显降低，差异有统计学意义(P<0.05)；而RLX、OPG和MMP-13的含量与模型组无统计学差异(P>0.05)；甲氨喋呤组中大鼠外周单核细胞中RANKL的mRNA水平的表达较模型组中降低，具有统计学差异(P<0.05)，而RXFP1和OPG的mRNA水平的表达与模型组无统计学差异(P>0.05)。
4.雷公藤多甙组与甲氨喋呤组比较
雷公藤多甙组大鼠血清中RLX、MMP9、MMP13和RANKL的含量、RANKL/OPG比值以及外周单核细胞中RXFP1和RANKL的mRNA相对表达均低于MTX组，但差异均无统计学意义(P>0.05)；雷公藤多甙组大鼠血清中OPG含量以及外周单核细胞中OPG的mRNA相对表达均高于MTX组，但差异均无统计学意义(P>0.05)。
5.直线相关性分析
RLX与MMP-9和RANKL呈正相关(r=0.428, P<0.05；r=0.416, P<0.05)。
结论
1.RLX可能通过上调MMP-9/MMP-13、RANKL/OPG途径导致CIA大鼠关节软骨破坏和骨质丢失；
2.雷公藤多甙减轻CIA大鼠关节软骨破坏和骨质丢失可能与抑制RLX/RXFP1、MMP-9/MMP-13、RANKL/OPG途径相关。
目的
研究中药雷公藤多甙治疗RA患者前后RLX/RXFP1、MMP-9/MMP-13和RANKL/OPG在血清以及分离的单核细胞中的表达水平，探讨它们参与关节炎软骨破坏和骨质丢失中的机制以及中药雷公藤多甙片治疗RA的药理机制。
方法
20例符合纳入标准的RA患者和30例健康人纳入本次研究，采集RA患者雷公藤多甙治疗前后以及正常人外周血标本以及相关临床资料。应用ELISA检测血清中RLX-2、MMP-9、MMP-13、OPG和RANKL的含量；采用荧光实时定量PCR方法检测外周单核细胞RXFP1、OPG和RANKL的mRNA的表达；比较雷公藤多甙片治疗前后RA患者的RLX/RXFP1、MMP-9/MMP-13和RANKL/OPG的表达有无差异。
结果
雷公藤多甙治疗前后RA患者血清RLX-2、MMP-9、MMP-13和RANKL，RANKL/OPG比值以及分离的外周单核细胞RXFP1和RANKL mRNA的表达具有统计学差异(P<0.05或P<0.01)。
结论
1.RLX-2可能通过上调MMP-9/MMP-13、RANKL/OPG途径参与RA患者关节软骨破坏和骨质丢失机制中；
2.雷公藤多甙缓解RA患者关节软骨破坏和骨质丢失可能与抑制RLX/RXFP1、MMP-9/MMP-13、RANKL/OPG途径相关。
目的
研究RLX/RXFP1、MMP-9/MMP-13和RANKL/OPG在不同疾病和证型的关节炎患者血清、外周单核细胞中的表达水平，以及探讨它们参与关节炎软骨破坏和骨质丢失的机制。
方法
共收集94例未治疗的关节炎患者（56例RA和38例OA）和30例正常人外周血标本。应用ELISA检测血清中RLX-2、MMP-9、MMP-13、OPG和RANKL的含量；通过抗凝血分离外周单核细胞，采用荧光实时定量PCR方法检测RXFP1、OPG和RANKL的mRNA的表达；比较RA、OA和正常组之间RLX-2/RXFP1、MMP-9/MMP-13和RANKL/OPG的表达有无差异。采用多元线性回归分析血清中RLX-2、MMP-9、MMP-13、RANKL和OPG的表达与RA患者以及OA患者的临床指标有无相关性；采用统计学直线相关性分析来分析血清中RLX-2与MMP-9和RANKL有无相关性。
结果
RA组和OA组血清RLX-2的含量均较正常组增高，差异有统计学意义(P<0.05或P<0.01)， RA组和OA组之间无统计学意义(P>0.05)；RA组和OA组外周血分离的单核细胞中RXFP1mRNA表达均较正常组增高，差异有统计学意义(P<0.05或P<0.01)，RA组和OA组之间无统计学意义(P>0.05)；RA组和OA组血清MMP-9的含量均较正常组增高，差异有统计学意义(P<0.05或P<0.01)， RA组和OA组之间有统计学意义(P<0.05)；RA组血清MMP-13的含量高于正常组但低于OA组，差异无统计学意义(P>0.05)；RA组血清OPG的含量较OA组和正常组低，差异无统计学意义(P>0.05)；RA组和OA组血清RANKL的含量较正常组高，差异有统计学意义(P<0.05或P<0.01)，RA组和OA组之间有统计学意义(P<0.05)；外周单核细胞中OPG和RANKL mRNA表达与血清中的表达呈现相同的特点；RA组和OA组血清RANKL/OPG比值均较正常组增高，差异有统计学意义(P<0.05或P<0.01)，RA组和OA组之间有统计学意义(P<0.05)；RA寒证或热证患者中血清MMP-9和RANKL以及外周单核细胞中RANKL的mRNA相对表达均高于OA寒证或热证患者，差异有统计学意义(P<0.05)。
多元线性回归分析示血清RLX-2和MMP-9表达均与ESR成正相关(β=0.417,t=2.126, P=0.039；β=0.435, t=2.328, P=0.042)；直线相关性分析示RLX-2与MMP-9和RANKL呈显著性正相关(r=0.417, P<0.01; r=0.423, P<0.01)。
结论
1.RLX-2可能通过上调MMP-9/MMP-13、RANKL/OPG途径参与RA和OA关节软骨破坏和骨质丢失机制中；
2.RLX-2和MMP-9可以反映RA患者的整体炎症水平。
Purpose
To 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.
Methods
CIA 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.
Results
1. Comparison between the normal group and the CIA model group
Serum 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 group
Serum 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 group
Serum 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 group
Serum 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 analysis
RLX was positively related to MMP-9and RANKL (r=0.428, P<0.05；r=0.416, P<0.05).
Conclusion
1. 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.
Purpose
To 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.
Methods
20cases 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.
Results
Serum 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).
Conclusion
1. 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.
Purpose
To 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.
Results
Serum 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).
Conclusion
1. 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.
通过建立CIA大鼠模型，研究RLX/RXFP1、MMP-9/MMP-13和RANKL/OPG在CIA大鼠模型血清和外周单核细胞中的表达水平，以及探讨RLX/RXFP1、MMP-9/MMP-13和RANKL/OPG在关节炎软骨破坏和骨质丢失中的机制和中药治疗中的作用。
方法
建立CIA大鼠模型，应用ELISA方法检测各组大鼠血清中RLX、MMP-9、MMP-13、OPG和RANKL的含量；采用荧光实时定量PCR方法检测各组大鼠的外周单核细胞中RXFP1、OPG和RANKL的mRNA的表达。比较CIA模型组和正常组之间RLX/RXFP1、MMP-9/MMP-13和RANKL/OPG的表达有无差异；比较药物治疗后与模型组之间RLX/RXFP1、MMP-9/MMP-13和RANKL/OPG的表达有无差异。
结果
1.正常组与CIA模型组比较
CIA模型组中大鼠血清RLX、MMP-9、RANKL的含量以及RANKL/OPG比值较正常组明显增高，差异有统计学意义(P<0.05)；而OPG和MMP-13的含量与正常组无统计学差异(P>0.05)；CIA模型组中大鼠的外周单核细胞中RXFP1和RANKL的mRNA水平的表达较正常组中增高，具有统计学差异(P<0.05)，而OPG的表达与正常组无统计学差异(P>0.05)。
2.雷公藤多甙组与模型组比较
雷公藤多甙组中大鼠血清中RLX、MMP-9、RANKL的含量以及RANKL/OPG比值较模型组明显降低，差异有统计学意义(P<0.05)；而OPG和MMP-13的含量与模型组无统计学差异(P>0.05)；雷公藤多甙组大鼠外周单核细胞中RXFP1和RANKL的mRNA水平的表达较模型组中降低，具有统计学差异(P<0.05)，而OPG的表达与模型组无统计学差异(P>0.05)。
3.甲氨喋呤组与模型组比较
甲氨喋呤组中大鼠血清中MMP-9和RANKL的含量以及RANKL/OPG比值较模型组明显降低，差异有统计学意义(P<0.05)；而RLX、OPG和MMP-13的含量与模型组无统计学差异(P>0.05)；甲氨喋呤组中大鼠外周单核细胞中RANKL的mRNA水平的表达较模型组中降低，具有统计学差异(P<0.05)，而RXFP1和OPG的mRNA水平的表达与模型组无统计学差异(P>0.05)。
4.雷公藤多甙组与甲氨喋呤组比较
雷公藤多甙组大鼠血清中RLX、MMP9、MMP13和RANKL的含量、RANKL/OPG比值以及外周单核细胞中RXFP1和RANKL的mRNA相对表达均低于MTX组，但差异均无统计学意义(P>0.05)；雷公藤多甙组大鼠血清中OPG含量以及外周单核细胞中OPG的mRNA相对表达均高于MTX组，但差异均无统计学意义(P>0.05)。
5.直线相关性分析
RLX与MMP-9和RANKL呈正相关(r=0.428, P<0.05；r=0.416, P<0.05)。
结论
1.RLX可能通过上调MMP-9/MMP-13、RANKL/OPG途径导致CIA大鼠关节软骨破坏和骨质丢失；
2.雷公藤多甙减轻CIA大鼠关节软骨破坏和骨质丢失可能与抑制RLX/RXFP1、MMP-9/MMP-13、RANKL/OPG途径相关。
目的
研究中药雷公藤多甙治疗RA患者前后RLX/RXFP1、MMP-9/MMP-13和RANKL/OPG在血清以及分离的单核细胞中的表达水平，探讨它们参与关节炎软骨破坏和骨质丢失中的机制以及中药雷公藤多甙片治疗RA的药理机制。
方法
20例符合纳入标准的RA患者和30例健康人纳入本次研究，采集RA患者雷公藤多甙治疗前后以及正常人外周血标本以及相关临床资料。应用ELISA检测血清中RLX-2、MMP-9、MMP-13、OPG和RANKL的含量；采用荧光实时定量PCR方法检测外周单核细胞RXFP1、OPG和RANKL的mRNA的表达；比较雷公藤多甙片治疗前后RA患者的RLX/RXFP1、MMP-9/MMP-13和RANKL/OPG的表达有无差异。
结果
雷公藤多甙治疗前后RA患者血清RLX-2、MMP-9、MMP-13和RANKL，RANKL/OPG比值以及分离的外周单核细胞RXFP1和RANKL mRNA的表达具有统计学差异(P<0.05或P<0.01)。
结论
1.RLX-2可能通过上调MMP-9/MMP-13、RANKL/OPG途径参与RA患者关节软骨破坏和骨质丢失机制中；
2.雷公藤多甙缓解RA患者关节软骨破坏和骨质丢失可能与抑制RLX/RXFP1、MMP-9/MMP-13、RANKL/OPG途径相关。
目的
研究RLX/RXFP1、MMP-9/MMP-13和RANKL/OPG在不同疾病和证型的关节炎患者血清、外周单核细胞中的表达水平，以及探讨它们参与关节炎软骨破坏和骨质丢失的机制。
方法
共收集94例未治疗的关节炎患者（56例RA和38例OA）和30例正常人外周血标本。应用ELISA检测血清中RLX-2、MMP-9、MMP-13、OPG和RANKL的含量；通过抗凝血分离外周单核细胞，采用荧光实时定量PCR方法检测RXFP1、OPG和RANKL的mRNA的表达；比较RA、OA和正常组之间RLX-2/RXFP1、MMP-9/MMP-13和RANKL/OPG的表达有无差异。采用多元线性回归分析血清中RLX-2、MMP-9、MMP-13、RANKL和OPG的表达与RA患者以及OA患者的临床指标有无相关性；采用统计学直线相关性分析来分析血清中RLX-2与MMP-9和RANKL有无相关性。
结果
RA组和OA组血清RLX-2的含量均较正常组增高，差异有统计学意义(P<0.05或P<0.01)， RA组和OA组之间无统计学意义(P>0.05)；RA组和OA组外周血分离的单核细胞中RXFP1mRNA表达均较正常组增高，差异有统计学意义(P<0.05或P<0.01)，RA组和OA组之间无统计学意义(P>0.05)；RA组和OA组血清MMP-9的含量均较正常组增高，差异有统计学意义(P<0.05或P<0.01)， RA组和OA组之间有统计学意义(P<0.05)；RA组血清MMP-13的含量高于正常组但低于OA组，差异无统计学意义(P>0.05)；RA组血清OPG的含量较OA组和正常组低，差异无统计学意义(P>0.05)；RA组和OA组血清RANKL的含量较正常组高，差异有统计学意义(P<0.05或P<0.01)，RA组和OA组之间有统计学意义(P<0.05)；外周单核细胞中OPG和RANKL mRNA表达与血清中的表达呈现相同的特点；RA组和OA组血清RANKL/OPG比值均较正常组增高，差异有统计学意义(P<0.05或P<0.01)，RA组和OA组之间有统计学意义(P<0.05)；RA寒证或热证患者中血清MMP-9和RANKL以及外周单核细胞中RANKL的mRNA相对表达均高于OA寒证或热证患者，差异有统计学意义(P<0.05)。
多元线性回归分析示血清RLX-2和MMP-9表达均与ESR成正相关(β=0.417,t=2.126, P=0.039；β=0.435, t=2.328, P=0.042)；直线相关性分析示RLX-2与MMP-9和RANKL呈显著性正相关(r=0.417, P<0.01; r=0.423, P<0.01)。
结论
1.RLX-2可能通过上调MMP-9/MMP-13、RANKL/OPG途径参与RA和OA关节软骨破坏和骨质丢失机制中；
2.RLX-2和MMP-9可以反映RA患者的整体炎症水平。
Purpose
To 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.
Methods
CIA 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.
Results
1. Comparison between the normal group and the CIA model group
Serum 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 group
Serum 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 group
Serum 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 group
Serum 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 analysis
RLX was positively related to MMP-9and RANKL (r=0.428, P<0.05；r=0.416, P<0.05).
Conclusion
1. 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.
Purpose
To 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.
Methods
20cases 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.
Results
Serum 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).
Conclusion
1. 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.
Purpose
To 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.
Results
Serum 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).
Conclusion
1. 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.
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29. Barrett AJ. Introduction to the history and classification of proteinases in: proteinases inmammalian cells and tissue. Elsevier/North-Holland, NY,1977,1-55.
30. Nagase H, Kashiwagi M. Aggrecanases and cartilage matrix degradation. Arthritis ResTher,2003,5(2):94-103.
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32. Nagase H, Woessner JF. Role of endogenous proteases in the degradation of the cartilagematrix. In: Joint cartlage degradation: Basic and clinical aspects. Marcel Dekker, NY,1993,159-185.
33. Hisaw F. Experimental relaxation of the pubic ligament of the guinea pig. Proc Soc ExPBiol Med,1926,23(23):661－663．
34. MarchenkoGN, Strongin AY. MMP-28, a new humanma-trixmetallo-protein ase with anunusual cysteine-switch sequence iswidely expressed in tumors. Gene,2001,265(122):87293.
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37. Murphy G, Nagase H. Reappraising metalloproteinases in rheumatoid arthritis andosteoarthritis: destruction or repair? Rheumatology,2008,4(3):128-135.
38. Konttinen YT, Ainola M, Valleala H, et al. Analysis of16different matrixmetalloproteinases (MMP-1to MMP-20) in the synovial membrane: different profiles intrauma and rheumatoid arthritis. Ann Rheum Dis,1999,58(11):691-697.
39. Elliott S, Hays E, Mayor M, et al. The triterpenoid CDDO inhibits expression of matrixmetalloproteinase-1, matrix metalloproteinase-13and Bcl-3in Primary humanchondrocytes. Arthritis Res Ther,2003,5(5): R285-291.
40. Bonassar LJ, Frank EH, Murray JC, et al. Changes in cartilage composition and physicalproperties due to stromelysin degradation. Arthritis Rheum,1995,38(2):173-183.
41. Mengshol JA, Mix KS, Brinckerhoff CE. Matrix metalloproteinases as therapeutic targetsin arthritic diseases: bull’s-eye or missing the mark? Arthritis Rheum,2002,46(1):13-20.
42. Smeets TJ, Kraan MC, Galjaard S, et al. Analysis of the cell infiltrate and expression ofmatrix metalloproteinases and granzyme B in paired synovialbiopsy specimens from thecartilage-pannus junction in patientswith RA. Ann Rheum Dis,2001,60(6):561-565.
43.魏平,付爽,王俊祥,等.基质金属蛋白酶3在类风湿关节炎中的表达及其意义.临床荟萃,2006,21(19):1389-1390.
44. Itoh T, UzukiM, Shimamura T, et al Dynamics of matrix metalloproteinase (MMP)-13inthe patients with rheumatoid arthritis. Ryumachi2002,42(1):60-69.
45.刘梦琼,林永前,王俏梅,等.类风湿关节炎和系统性红斑狼疮患者血清基质金属蛋白酶2、9检测的意义.检验医学,2009,24(12):883-885.
46.范慧洁,戴如春,廖二元. OPG/RANK/RANKL系统与骨折和类风湿性关节炎.国际内分泌代谢杂志,2006,26(4):241-244.
47. Dougall WC, Glaccum M, Charrier K, et al. RANK is essential for osteoclast and lymphnode development. Genes Dev,1999,13(18):2412-2424.
48. Bucay N, Sarosi I, Dunstan CR, et al. osteoprotegerin-deficient mice develop early onsetosteoporosis and arterial calcification. Genes Dev,1998,12(9):1260-1268.
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