白芍总苷调控AMPK/RANKL/OPG轴治疗CIA模型大鼠骨破坏的作用研究

Effect of Total Glucoside of Paeoniae Radix Alba on Bone Destruction in CIA Model Rats by Regulating AMPK/RANKL/OPG Axis

目的:探讨白芍总苷调控AMPK/RANKL/OPG轴治疗CIA大鼠骨破坏的作用研究。方法:60只造模成功的雌性Wistar大鼠随机分为模型组、AICAR组、来氟米特组及白芍总苷高、中、低剂量组,每组10只,另选取10只健康雌性Wistar大鼠作为正常对照组。监测大鼠体质量及足趾肿胀度变化,HE染色观察大鼠膝关节滑膜组织病理学变化,ELISA法检测血清TNF-α、IL-1α、IL-6、IL-10水平;RT-PCR和Western Blot检测滑膜组织AMPK、OPG、RANKL、TRAP mRNA和(或)蛋白表达,彩色B超及小动物断层扫描(Micro-CT)观察大鼠关节软骨及软骨下骨质破坏情况。结果:与模型组比较,白芍总苷可有效缓解关节肿胀,呈剂量依赖性降低CIA模型大鼠血清中TNF-α、IL-1α、IL-6水平及滑膜RANKL、TRAP mRNA及蛋白表达,升高滑膜OPG、p-AMPK mRNA和(或)蛋白表达(P<0.05~P<0.001),中剂量的作用与AICAR相当。来氟米特提高AMPK信号相关mRNA表达的作用不明显。结论:白芍总苷可一定程度上改善CIA大鼠关节破坏,其可能是通过调控AMPK/RANKL/OPG信号通路来发挥RA的骨保护作用。

Objective:To explore the effect of total glucoside of Paeoniae Radix Alba on bone destruction in CIA model rats by regulating AMPK/RANKL/OPG axis.Methods:Sixty successful female Wistar rats were randomly divided into model group,AICAR group,leflunomide group,high,medium and low dose of total glucoside of Paeoniae Radix Alba groups with 10 in each group.Another 10 healthy female Wistar rats were selected as normal control group.After 4 weeks of administration,the changes of body mass and toe swelling were monitored,The histopathological changes of knee sunovium were observed by HE staining.Serum TNF-α,IL-1α,IL-6 and IL-10 levels were detected by ELISA.The mRNA and/or protein expressions of AMPK,OPG,RANKL and TRAP were detected by RT-PCR and Western Blot.The destruction of articular cartilage and subchondral bone in rats was observed by color B-ultrasound and Micro-CT.Results:Compared with the model group,total glucoside of Paeoniae Radix Alba effectively alleviated joint swelling.In a dose-dependent manner,total glucoside of Paeoniae Radix Alba decreased the levels of TNF-α,IL-1αand IL-6 in serum and the expressions of synovial RANKL and TRAP mRNA and proteins in CIA model rats,and the expressions of OPG,p-AMPK mRNA and/or proteins in synovial membrane was increased(P<0.05 to P<0.001),and the effect of medium dose was similar to that of AICAR.Leflunomide did not significantly increase the mRNA expression of AMPK signal.Conclusion:The total glucosides of Paeoniae Radix Alba can ameliorate joint destruction in CIA rats to a certain extent,which may play the bone protective role of RA by regulating AMPK/RANKL/OPG signaling pathway.