紫花牡荆素抑制软骨下骨破坏延缓骨关节炎进展的研究

Casticin attenuates early osteoarthritis progression by inhibiting subchondral bone destruction

目的探讨紫花牡荆素是否可延缓骨关节炎(OA)的进展。方法选取18只清洁级C57bl/6j小鼠,6~8周龄,体重18~22g。将其随机分为3组(n=6):假手术组、前交叉韧带切除(ACLT)溶媒组(OA模型组)、ACLT紫花牡荆素组(OA治疗组)。假手术组小鼠切开膝关节囊,ACLT溶媒组和ACLT紫花牡荆素组小鼠均行右膝ACLT。假手术组、ACLT溶媒组给予等量的Tween-80溶液灌胃;ACLT紫花牡荆素组以20mg/kg剂量每日灌胃给药1次。Micro-CT评估软骨下骨改变,抗酒石酸酸性磷酸酶TRAP染色评估软骨下骨破骨细胞数目,免疫组化评估软骨下骨NOX4的表达情况。使用HE、番红-O绿染色评估软骨退变程度并对软骨退变行Mankin评分。结果与假手术组相比,ACLT溶媒组小鼠软骨下骨骨密度、骨体积分数、骨小梁厚度明显下降,骨小梁间隙缩小,TRAP染色、NOX-4免疫组化染色阳性率和软骨损伤Mankin评分明显上升,差异均有统计学意义(P<0.05)。与ACLT溶媒组相比,ACLT紫花牡荆素组小鼠软骨下骨骨密度、骨体积分数上升,软骨下骨骨小梁间隙、TRAP染色、NOX-4免疫组化染色阳性率和软骨损伤Mankin评分明显降低,差异均有统计学意义(P<0.05)。结论紫花牡荆素可抑制OA模型软骨下骨中NOX4的表达,抑制破骨细胞形成,延缓软骨下骨、软骨退变及OA进展,为OA治疗带来了新思路。

Objectives To investigate whether osteoarthritis(OA)progression can be delayed by casticin in rodent models of anterior cruciate ligament transection(ACLT).Methods Eighteen 2-month-old male C57BL/6J mice were randomised into 3 even groups(n=6)subjected respectively to sham-operation,ACLT-vehicle-treatment and ACLT-casticin-treatment.The knee capsule was dissected in the sham-operation group and ACLT on the right knee was conducted in the ACLT-vehicle-treatment and ACLT-casticin-treatment groups.Intragastric administration of the same amount of Tween-80 solution was conducted for the sham-operation and ACLT-vehicle-treatment groups;Intragastric administration of casticin of 20 mg/kg was conducted once per day for the ACLT-casticin-treatment group.Bone micro CT(μCT)was quantitated to detect alterations in microarchitecture of femoral condyle subchondral bone.Tartrate resistant acid phosphatase(TRAP)stain and NOX4 immunostaining were conducted to detect relative proteins and the osteoclast changes on the subchondral bone.Articular cartilage degeneration was graded using HE and safranin O-green staining and the Mankin score criteria.Results Compared with the the sham-operation group,the subchondral bone density,trabecular bone volume fraction and trabecular thickness were decreased,and the trabecular space,positive rates of TRAP stain and NOX4 immunostaining and Mankin scores were increased in the ACLT-vehicle-treatment group.All the above comparisons were statistically significant(P<0.05).Compared with ACLT-vehicle-treatment group,the subchondral bone density and trabecular bone volume fraction were increased,and the trabecular space,positive rates of TRAP stain and NOX4 immunostaining and Mankin scores were decreased in the ACLT-casticin-treatment group.All the above comparisons were statistically significant(P<0.05).Conclusion As casticin may attenuate early OA progression by inhibiting NOX4 activity in subchondral bone and formation of osteoclasts,it may be a new clue to preventive therapy for OA.