同源点针刺治疗骨质疏松症大鼠的作用机制
The mechanism of homologous point acupuncture in the treatment of osteoporosis in rats
  
DOI:10.3969/j.issn.1006-7108.2025.07.003
中文关键词:  针刺  绝经后骨质疏松症  同源点疗法  OPG/RANKL信号通路
英文关键词:acupuncture  postmenopausal osteoporosis  homologous point therapy  OPG/RANKL pathway
基金项目:湖北省自然科学基金(2023AFD141)
作者单位
黄觅1 彭高高2 刘常鲲2 吴汉卿2 胡昭端2 彭锐1,2* 1.武汉市中西医结合医院湖北 武汉 430030 2.湖北中医药大学湖北 武汉 430065 
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中文摘要:
      目的 研究同源点针刺疗法对绝经后骨质疏松症(postmenopausal osteoporosis,PMOP)大鼠的疗效及对骨保护素(OPG)/核因子-κB受体活化因子配体(RANKL)信号通路的影响。方法 选择SPF级雌性SD大鼠36只,随机分为对照组、模型组、假针刺组、低频治疗组、中频治疗组及高频治疗组,每组6只。除对照组外,其他5组大鼠均采用去除双侧卵巢法建立PMOP模型。造模成功后进行为期12周的同源点针刺及假针刺干预。干预结束后采用酶联免疫吸附测定法(ELISA)检测大鼠血清I型胶原羧基末端肽(CTX-I)、I型前胶原氨基端前肽(PINP)、骨特异性碱性磷酸酶(BALP)、肿瘤坏死因子-α(TNF-α)、骨形态发生蛋白-2(BMP-2)含量;采用显微计算机断层成像(Micro-CT)分析股骨骨密度(BMD)、骨小梁厚度(Tb.Th)、骨小梁分离度(Tb.Sp)、骨小梁数量(Tb.N);采用苏木精-伊红染色(HE)观察股骨组织形态变化;采用免疫组织化学(IHC)检测大鼠股骨组织中OPG、RANKL、BMP-2及基质金属蛋白酶-2(MMP-2)蛋白表达水平;采用免疫印迹(WB)检测大鼠股骨组织中OPG、RANKL、TNF-α、BMP2及MMP2蛋白表达水平。结果 与对照组比较,模型组大鼠血清PINP、BALP、股骨BMD、Tb.Th、Tb.N、OPG及MMP-2蛋白水平明显下降(P<0.05),血清CTX-I、TNF-α、BMP-2、股骨Tb.Sp、RANKL及BMP-2蛋白水平明显上升(P<0.01),HE染色及Micro-CT显示骨小梁数量减少。与模型组比较,低频治疗组血清CTX-I、BMP-2、TNF-α、股骨Tb.Sp、RANKL及BMP-2蛋白水平明显下降(P<0.05),血清PINP、BALP、股骨BMD、Tb.Th、Tb.N、OPG及MMP-2蛋白水平明显上升(P<0.05),中频治疗组血清BMP-2、TNF-α、RANKL及BMP-2蛋白水平明显下降(P<0.05),BMD、Tb.Th、OPG及MMP-2蛋白水平明显上升(P<0.05),高频治疗组血清CTX-I、BMP-2、TNF-α、Tb.Sp、Rankl蛋白水平明显下降(P<0.05),Tb.N、Tb.Th及OPG蛋白水平明显上升(P<0.05),HE染色及Micro-CT显示低频治疗组骨小梁结构较致密。结论 同源点针刺治疗能够改善PMOP大鼠的骨代谢,其中低频治疗组改善效果更明显,可能是通过调控OPG/RANKL信号通路发挥作用。
英文摘要:
      Objective To explore the efficacy of homologous point acupuncture therapy on postmenopausal osteoporosis (PMOP) model in ovariectomized rats and its effect on osteoprotegerin (OPG)/receptor activator of nuclear factor-κB ligand (RANKL) signaling pathway. Methods Thirty-six SPF-grade female SD rats were selected and randomly divided into the control group, model group, sham acupuncture group, low-frequency (LF) treatment group, medium-frequency (MF) treatment group, and high-frequency (HF) treatment group, with 6 rats in each group. Except for the control group, the osteoporosis model was established using the ovary removal method in the other five groups. After successful modeling, homologous point acupuncture therapy and sham acupuncture interventions were carried out for 12 weeks. At the end of the intervention, CTX-I, PINP, BALP, TNF-α, and BMP-2 in rat serum were detected using ELISA. BMD of the femur, Tb.Th, Tb.Sp, and Tb.N were analyzed using micro-CT. The femoral tissue morphology was observed with HE staining. The levels of OPG, RANKL, BMP-2, and MMP-2 protein expression in the femur were detected using IHC. WB was used to detect OPG, RANKL, TNF-α, BMP2, and MMP2 protein expression levels in the rat femur. Results Compared to those in the control group, the serum levels of PINP, BALP, BMD of the femur, Tb.Th, Tb.N, OPG, and MMP-2 proteins decreased significantly (P<0.05), and the serum levels of CTX-I, BMP-2, TNF-α, Tb.Sp, and RANKL and BMP-2 proteins increased significantly in the model group (P<0.01). HE staining and micro-CT results showed a decrease in the number of bone trabeculae. Compared to those in the model group, the serum levels of CTX-I, TNF-α,BMP-2, Tb.Sp, and RANKL and BMP-2 proteins decreased significantly in the LF treatment group (P<0.05), and the serum levels of PINP, BALP, BMD of the femur, Tb.Th, Tb.N, and OPG and MMP-2 proteins increased significantly in the LF treatment group (P<0.05). The serum levels of BMP-2, TNF-α, RANKL, and BMP-2 protein decreased significantly in the MF treatment group (P<0.05). BMD of the femur, Tb.Th, and OPG and MMP-2 protein levels increased significantly in the MF treatment group (P<0.05). The serum levels of CTX-I, BMP-2, TNF-α, Tb.Sp, and RANKL protein decreased significantly in the HF treatment group (P<0.05). Tb.N, Tb.Th, and OPG proteins increased significantly (P<0.05). HE staining and micro-CT results showed denser bone trabecular structure in the LF treatment group. Conclusion Homologous point acupuncture therapy improves the bone metabolism in PMOP rats. The improvement effect is more obvious in the LF treatment group, which may work through the regulation of OPG/RANKL signaling pathway.
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