摘要
目的 观察骨痹通消颗粒对激素性股骨头坏死模型小鼠的治疗作用,并从核因子-κB受体活化因子配体/核因子-κB受体活化因子/骨保护素(RANKL/RANK/OPG)信号轴探讨其作用机制.方法 采取腹腔注射脂多糖和臀肌注射醋酸泼尼松龙复制激素性股骨头缺血坏死(SONFH)动物模型,经核磁共振鉴定后,将模型复制成功的60只小鼠分成模型(MC)组、骨痹通消颗粒(GBTX)组及通络生骨胶囊(PC)组,每组20只,另设12只正常小鼠作为空白对照(NC)组.分别予以对应组灌胃相应药物12周后麻醉状态下取材,观察各组小鼠治疗前后的一般行为;酶联免疫吸附试验(ELISA)检测各组小鼠的骨碱性磷酸酶(BALP)和I型氨基端延长肽(PINP)含量;Western blotting和实时荧光定量聚合酶链反应(qRT-qPCR)检测各组RANK、RANKL、OPG、磷脂酰肌醇特异性磷脂酶Cγ2(PLCγ2)、组织蛋白酶K(CTSK)、抗酒石酸酸性磷酸酶(TRAP)的蛋白和基因表达.结果 核磁共振显示,模型复制成功后的小鼠左侧髋部呈高信号状态.与NC组比较,MC组BALP、PINP水平,以及OPG蛋白和基因相对表达量均降低(P<0.05),RANK、RANKL、PLCγ2、CTSK、TRAP蛋白和基因相对表达量均升高(P<0.05);与MC组比较,GBTX组和PC组BALP、PINP水平,以及OPG蛋白和基因相对表达量均升高(P<0.05),RANK、RANKL、PLCγ2、CTSK、TRAP蛋白和基因相对表达量均下降(P<0.05);与GBTX组比较,PC组OPG、RANK、RANKL、CTSK、TRAP蛋白和基因相对表达量,以及PLCγ2蛋白相对表达量差异均无统计学意义(P>0.05),而PC组PLCγ2基因相对表达量较GBTX组下降(P<0.05).结论 骨痹通消颗粒可能通过上调OPG表达,抑制RANK、RANKL、PLCγ2、CTSK和TRAP表达,从而改善股骨头坏死情况.
Abstract
Objective To observe the therapeutic effects of Gubitongxiao granules on hormone-induced femoral head necrosis(SONFH)in mice and explore its mechanism of action based on the receptor activator of nuclear factor-κB ligand/receptor activator of nuclear factor-κB/osteoprotegerin(RANKL/RANK/OPG)signaling pathway.Methods A SONFH animal model was established by intraperitoneal injection of lipopolysaccharide and intramuscular injection of methylprednisolone acetate into the gluteus of 60 mice.After confirmation by magnetic resonance imaging,the mice were divided into a model group(MC),a Gubitongxiao granules group(GBTX),and a Tongluo Shenggu Gelatin Capsules group(PC),with 20 mice in each group.An additional 12 normal mice were used as a blank control group(NC).Corresponding drugs were administered orally to each group for 12 weeks.Subsequently,under anesthesia,samples were collected,and the general behaviors of the mice were observed.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of bone alkaline phosphatase(BALP)and type I procollagen N-terminal propeptide(PINP)in the mice.Western blotting and real-time quantitative polymerase chain reaction(qRT-qPCR)were used to measure the protein and gene expression levels of RANK,RANKL,OPG,phospholipase Cγ2(PLCγ2),cathepsin K(CTSK),and tartrate-resistant acid phosphatase(TRAP)in each group.Results Magnetic resonance imaging showed a high signal state in the left hip of the mice after successful model replication.Compared with the NC group,the MC group showed decreased levels of BALP,PINP,OPG protein,and gene expression(P<0.05),and increased levels of RANK,RANKL,PLCγ2,CTSK,and TRAP protein and gene expression(P<0.05).Compared with the MC group,the GBTX and PC groups showed increased levels of BALP,PINP,OPG protein,and gene expression(P<0.05),and decreased levels of RANK,RANKL,PLCγ 2,CTSK,and TRAP protein and gene expression(P<0.05).Compared with the GBTX group,the PC group showed no significant differences in OPG,RANK,RANKL,CTSK,and TRAP protein and gene expression(P>0.05),while the gene expression of PLCγ2 in the PC group was lower than that in the GBTX group(P<0.05).Conclusion Gubitongxiao granules may improve femoral head necrosis by upregulating OPG expression,inhibiting RANK,RANKL,PLCγ2,CTSK,and TRAP expression,thereby ameliorating the condition.
基金项目
国家自然科学基金面上项目(81373664)
安徽省自然科学基金面上项目(KJ2020A0403)
维普
万方数据