双固针法通过调节肠道菌群与雌二醇代谢改善去卵巢大鼠骨质疏松

The Shuanggu Needling Method Ameliorates Osteoporosis in Ovariectomized Rats by Regulating Gut Microbiota and Estradiol Metabolism

摘要: 目的：绝经后骨质疏松症(PMOP)是严重的全球性健康问题，探索安全有效的防治策略至关重要。本研究基于“肠–骨轴”理论，旨在系统阐释“双固一通”针法中“双固”法(电针关元、足三里穴)对去卵巢(OVX)大鼠PMOP模型的治疗效应及其多维度作用机制。方法：将80只雌性SD大鼠随机分为假手术组(Sham)、模型组(OVX)、双固针4周治疗组(OVX + EA-4W)及8周治疗组(OVX + EA-8W)。通过综合运用双能X线骨密度检测、骨组织形态计量学及酶联免疫吸附测定(ELISA)等技术，多尺度评价双固针法的干预效果。结果：与OVX组相比，双固针干预能显著提升大鼠腰椎与股骨的骨密度(BMD)和骨矿物质含量(BMC) (P < 0.01)，改善骨小梁微结构，上调血清骨钙素(BGP)水平并下调I型胶原羧基端肽(β-CTX)水平(P < 0.05)。肠道菌群分析表明，双固针法可有效逆转OVX诱导的菌群失调，显著提高乳酸菌、拟杆菌等有益菌的相对丰度，降低大肠杆菌志贺菌等潜在致病菌的比例，并显著改善菌群Alpha多样性。结论：双固针法可能通过重塑“肠道菌群–代谢物–内分泌”稳态网络，激活成骨信号通路并抑制破骨活性，从而多靶点防治PMOP。本研究为针灸疗法的现代化阐释提供了坚实的科学依据。

Abstract: Objective: Postmenopausal osteoporosis (PMOP) is a severe global health issue, and exploring safe and effective prevention and treatment strategies is crucial. Based on the “gut-bone axis” theory, this study aims to systematically elucidate the therapeutic effects and multi-dimensional mechanisms of the “Shuanggu” method (electroacupuncture at Guanyuan [CV4] and Zusanli [ST36] acupoints)—a component of the “Shuanggu-Yitong” acupuncture technique—on the PMOP model in ovariectomized (OVX) rats. Methods: Eighty female Sprague-Dawley (SD) rats were randomly divided into four groups: sham operation group (Sham), ovariectomized model group (OVX), 4-week treatment group with Shuanggu acupuncture (OVX + EA-4W), and 8-week treatment group (OVX + EA-8W). The intervention effects of the Shuanggu Needling Method were evaluated using multi-scale techniques, including dual-energy X-ray absorptiometry (DXA) for bone mineral density, bone histomorphometry, and enzyme-linked immunosorbent assay (ELISA). Results: Compared with the OVX group, Shuanggu acupuncture significantly increased bone mineral density (BMD) and bone mineral content (BMC) in the lumbar vertebrae and femur (P < 0.01), improved trabecular microstructure, upregulated serum bone gla protein (BGP) levels, and downregulated C-terminal telopeptide of type I collagen (β-CTX) levels (P < 0.05). Gut microbiota analysis revealed that the Shuanggu Needling Method effectively reversed OVX-induced dysbiosis, significantly increased the relative abundance of beneficial bacteria such as Lactobacillus and Bacteroides, reduced the proportion of potential pathogens like Escherichia coli-Shigella, and markedly improved microbial alpha diversity. Conclusion: The Shuanggu Needling Method may prevent and treat PMOP through multi-target mechanisms, including reshaping the homeostatic network of “gut microbiota-metabolites-endocrine”, activating osteogenic signaling pathways, and inhibiting osteoclast activity. This study provides a solid scientific basis for the modern interpretation of acupuncture therapy.