摘要
目的 探讨1,25-二羟基维生素D_(3)[1,25-dihydroxy vitamin D3,1,25(OH)_(2)D_(3)]对胶原诱导的关节炎(collagen-induced arthritis,CIA)模型小鼠关节滑膜炎症和骨破坏的抑制作用及机制。方法 30只DBA/1J小鼠被均分为3组:对照组(CON)、模型组(CIA)和模型干预组(VD)。CIA组和VD组小鼠建立CIA模型,用1,25(OH)_(2)D_(3)对VD组CIA模型进行治疗。用游标卡尺测量小鼠后足爪的肿胀程度。用Micro-CT测量小鼠后足爪的骨密度。酶联免疫吸附剂测定法检测小鼠血清1,25(OH)_(2)D_(3)和炎症细胞因子(IL-1β、IL-6、TNF-α)的水平。流式细胞术检测小鼠脾脏Th17细胞和Treg细胞的比例。采用苏木精-伊红染色观察膝关节的病理特征。Western blot检测小鼠踝关节的蛋白OPG、RANKL、NF-κB和VDR的表达水平。结果 1,25(OH)_(2)D_(3)并没有降低CIA模型的发病率,但却明显缓解了关节炎症。VD组的脚掌厚度、关节炎评分以及膝关节HSS评分均较CIA组明显降低(P均<0.05)。CIA组的骨破坏程度更加明显,BMD及BV/TV明显低于VD组(P均<0.05)。VD组的IL-1β、IL-6、TNF-α水平和Th17/Treg细胞比例明显低于CIA组(P均<0.05)。与CIA组相比,VD组踝关节的VDR和OPG蛋白水平明显增加(P均<0.05),NF-κB和RANKL蛋白水平明显低于CIA组(P均<0.05),RANKL/OPG的比例也明显低于CIA组(P<0.05)。结论 1,25(OH)_(2)D_(3)可能通过调节Th17/Treg比值和RANKL/OPG轴缓解CIA模型小鼠的关节滑膜炎症并抑制骨破坏。
Objective To investigate the mechanisms of 1,25(OH)_(2)D_(3) on joint inflammation and bone destruction in a mouse model of collagen-induced arthritis(CIA).Methods Thirty DBA/1J mice were equally allocated into control,CIA and intervention(VD) groups.The CIA model was established in the mice in the CIA group and VD group,meanwhile the VD group was treated with 1,25(OH)_(2)D_(3).The thickness of the hindfoot paws was measured with vernier caliper.Micro-CT scanning was used to measure bone mineral density(BMD) of the hindfoot paw,ELISA was employed to detect the serum levels of 1,25(OH)_(2)D_(3) and proinflammatory cytokines(IL-1β,IL-6,TNF-α),and flow cytometry was performed to calculate the ratio of Th17 to Treg cells in the spleen.The pathological changes of the knee joint were observed with HE staining.Western blotting was applied to detect the expression levels of osteoprotegerin(OPG),receptor activator of nuclear factor-κB ligand(RANKL),NF-κB,and vitamin D receptor(VDR) in mouse ankle joints.Results 1,25(OH)_(2)D_(3) did not reduce the incidence of CIA in the of mice in the VD group,but significantly reduced joint inflammation.Obviously decreased thickness of the hindfoot paw,arthritis score,and knee histological synovitis score(HSS) were observed in the VD group than the CIA group(all P<0.05).The CIA group presented more serious bone destruction and lower BMD and bone volume fraction(BV/TV) when compared with the VD group(both P<0.05).The serum levels of IL-1β,IL-6 and TNF-α and ratio of Th17/Treg cells in the VD group were statistically lower than those in the CIA group(all P<0.05).Compared with the CIA group,the VD group had notably higher protein levels of VDR and OPG,lower levels of NF-κB and RANKL,and reduced RANKL/OPG ratio in the ankle joint(all P<0.05).Conclusion 1,25(OH)_(2)D_(3) may alleviate arthritic inflammation and suppress bone destruction in the mouse CIA model via regulating the Th17/Treg ratio and the RANKL/OPG axis.
作者
齐坤
刘云
唐滔
罗晰文
张宇
张志勇
安云飞
唐雪梅
QI Kun;LIU Yun;TANG Tao;LUO Xiwen;ZHANG Yu;ZHANG Zhiyong;AN Yunfei;TANG Xuemei(Department of Rheumatology&Immunology,National Clinical Research Center for Child Health and Disorders,Key Laboratory of Child Development and Disorders of Ministry of Education,Chongqing Key Laboratory of Child Infection and Immunity,Children's Hospital of Chongqing Medical University,Chongqing,400014,China)
出处
《陆军军医大学学报》
CAS
CSCD
北大核心
2023年第15期1656-1665,共10页
Journal of Army Medical University
基金
国家重点研发计划(2021YFC2702003)。
