盘龙七片调节骨保护素/核因子κB受体活化因子配体信号通路对去卵巢骨质疏松大鼠的影响

Study of Efficacy of Panlongqi Tablet Modulated OPG/RANKL Pathway in Ovariectomized Rats with Osteoporosis

目的:探讨盘龙七片对去卵巢骨质疏松大鼠骨保护素(OPG)/核因子κB受体活化因子配体(RANKL)信号通路的影响。方法:75只雌性SD大鼠随机分为假手术组、模型组、阳性药物对照组(17β-雌二醇,1 mg/kg)、盘龙七片低剂量组(0.26 g/kg)、盘龙七片高剂量组(1.29 g/kg),每组15只,除假手术组外,其余各组大鼠建立去卵巢骨质疏松模型,分组给予相应药物处理后,苏木精-伊红(HE)染色法染色观察各组大鼠股骨并成像;通过双能X线骨密度分析仪对大鼠离体股骨密度进行检测;ELISA测定血清β-胶原降解产物(β-CTX)和骨碱性磷酸酶(BALP)水平;Western Blot测定左侧胫骨OPG、RANKL蛋白相对表达水平。结果:模型组左侧股骨远端骨小梁稀疏、结构不完整、明显变薄、排列紊乱;盘龙七片低剂量组左侧股骨远端骨小梁有所改善,盘龙七片高剂量组与阳性药物组左侧股骨远端骨小梁形态结构明显改善。与假手术组相比,模型组大鼠BMC、最大载荷与弹性模量、BALP水平及OPG蛋白表达水平显著降低,差异有统计学意义(P<0.05),血清中β-CTX水平及RANKL蛋白水平显著升高,差异有统计学意义(P<0.05);与模型组相比,盘龙七片低、高剂量组及阳性药物组大鼠BMC、最大载荷与弹性模量、BALP水平及OPG蛋白表达水平显著升高,差异有统计学意义(P<0.05),血清中β-CTX水平及RANKL蛋白水平显著降低,差异有统计学意义(P<0.05)。结论:盘龙七片可能通过抑制RANKL水平,上调OPG表达,改善去卵巢大鼠的骨质疏松症状。

Objective:To investigate the efficacy of Panlongqi tablet on osteoprotegerin(OPG)/receptor activator of nuclear factor-κB ligand(RANKL)signaling pathway in ovariectomized osteoporosis rats.Methods:A total of 75 female SD rats were randomly divided into sham operation group,model group,positive drug control group(17β-estradiol,1 mg/kg),Panlongqi tablet low-dose group(0.26 g/kg)and Panlongqi tablet high-dose group(1.29 g/kg),with 15 rats in each group.Excepting the sham operation group,ovariectomized osteoporosis models were established in other groups.After treatment with corresponding drugs in groups,the femurs were observed and imaged by Hematoxylin Eosin staining(HE).The density of femur was detected by dual energy X-ray absorptiometry.The expressions ofβ-cross-linked c-telopeptide of typeⅠcollagen(β-CTX)and bone alkaline phosphatase(BALP)in serum were detected by ELISA,and the related expression level of OPG/RANKL protein in left tibia was measured by Western Blot.Results:The trabecular bones of the distal femur on the left side in the model group were sparse,structurally incomplete,obviously thinned and disorderly arranged.The trabecular bone of the left distal femur in the low-dose tablet group was improved,and the trabecular bone morphology and structure of the left distal femur in the high-dose Panlongqi tablet group and the positive drug group were significantly improved.Compared with the sham operation group,BMC,maximum load and elastic modulus,BALP levels and OPG protein expression levels in the model group were significantly reduced(P<0.05),andβ-CTX levels and RANKL protein levels in serum were significantly increased(P<0.05).Compared with the model group,the BMC,maximum load and elastic modulus,BALP level and OPG protein expression level of Panlongqi tablet low,high dose group and positive drug group increased significantly(P<0.05).Theβ-CTX level and RANKL protein level in serum were significantly reduced(P<0.05).Conclusion:Panlongqi tablet may inhibit the level of RANKL and up-regulate the expression of OPG to improve the osteoporosis of ovariectomized rats.