中华中医药学刊

基于16S rDNA测序探讨加味活络效灵丹对激素性股骨头坏死大鼠“肠-骨”轴的影响

PDF下载

张江 ¹^,²^,³ , 胡宇晴 ¹ , 张诗宇 ² , 董野 ¹ , 侯德才 ²

中华中医药学刊 | 国家项目点击 2026,44(9): 53-58

收起

中华中医药学刊 |国家项目点击 2026 , 44 (9) : 53 -58

基于16S rDNA测序探讨加味活络效灵丹对激素性股骨头坏死大鼠“肠-骨”轴的影响

全屏

张江¹^,²^,³, 胡宇晴¹, 张诗宇², 董野¹, 侯德才²

作者信息

^1.辽宁中医药大学，辽宁　沈阳　110847

^2.辽宁中医药大学附属医院，辽宁　沈阳　110032

^3.辽宁中医药大学中医脏象理论及应用教育部重点实验室，辽宁　沈阳　110847

通讯作者:

侯德才（1961-），男，辽宁沈阳人，教授、主任医师，博士研究生导师，博士，研究方向：中医药防治股骨头坏死。E-mail：lnzyhdc@163.com。

作者简介:

张江（1988-），男，辽宁辽阳人，主治医师，博士在读，研究方向：中医药防治股骨头坏死。

Effect of Modified Huoluo Xiaoling Dan (加味活络效灵丹) on “Gut-osteoporosis” Axis of Rats with Hormone Induced Femoral Head Necrosis Based on 16S rDNA Sequencing

Jiang ZHANG¹^,²^,³, Yuqing HU¹, Shiyu ZHANG², Ye DONG¹, Decai HOU²

Affiliations

^1.Liaoning University of Traditional Chinese Medicine, Shenyang 110847, Liaoning, China

^2.Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang 110032, Liaoning, China

^3.Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications, Liaoning University of Traditional Chinese Medicine, Shenyang 110847, Liaoning, China

出版时间: 2026-09-10 doi: 10.13193/j.issn.1673-7717.2026.09.010

文章导航

摘要

收起

目的

基于“肠-骨”轴理论和16S rDNA技术，探究加味活络效灵丹对激素性股骨头坏死大鼠骨组织修复的作用与机制，及其肠道菌群结构的影响。

方法

采用随机数字表法，将24只SPF级雄性SD大鼠分为正常组（n=8）、模型组（n=8）、治疗组（n=8）。模型组和治疗组采用臀肌交替注射甲泼尼龙琥珀酸钠的造模方法，治疗组予加味活络效灵丹（制剂药液）灌胃。对取材的大鼠股骨样品，采用HE染色并计算空骨陷窝率，Micro-CT三维重建分析测量区域骨体积分数（BV/TV）、骨小梁数量（Tb. N）、骨小梁厚度（Tb. Th）、骨小梁分离度（Tb. Sp）等骨组织学参数，免疫组化检测骨组织修复标记物BMP、OPG、Runx2表达；对其粪便样品，采用16S rDNA技术进行肠道菌群测序与ASV分析，α多样性和β多样性分析肠道菌群物种多样性，LEfSe分析肠道菌群物种差异性在科水平和属水平上的改变。

结果

加味活络效灵丹可显著改善SANFH大鼠股骨头空骨陷窝率（P<0.05）及BV/TV、Tb. N、Tb. Th、Tb. Sp等骨组织学参数（P<0.01），并显著上调骨组织修复标记物BMP、OPG、Runx2表达水平（P<0.01）。肠道菌群测序结果显示，α多样性分析说明3组大鼠肠道菌群结构的物种多样性和丰富度存在明显差异；β多样性分析显示治疗组菌群结构与模型组存在显著分离，与空白组相近，组间差异大于组内，且差异明显。LEfSe分析发现在科水平和属水平上，模型组大鼠肠道菌群呈现显著的“促炎型”特征，治疗组大鼠肠道菌群呈现“抗炎-促修复”特征。

结论

加味活络效灵丹对SANFH大鼠骨组织修复作用与其调控肠道菌群密切相关，干预药物有效重塑了激素扰动的肠道微生态，进一步证实菌群重塑对“肠-骨”轴的双向调节，起到防治SANFH的作用。

关键词

激素性股骨头坏死 / 加味活络效灵丹 / 肠道菌群 / 骨组织修复 / “肠-骨”轴

Abstract

收起

Objective

Based on the theory of the “gut-osteoporosis” axis and 16S rDNA technology, to explore the effect and mechanism of Modified Huoluo Xiaoling Dan (加味活络效灵丹) on the repair of bone tissue in the rats with steroid-induced avascular necrosis of the femoral head (SANFH), as well as its influence on the structure of the gut microbiota.

Methods

Using the random number table method, 24 SPF-grade male SD rats were divided into a normal group (n=8), a model group (n=8) and a treatment group (n=8). In the model group and the treatment group, the method of alternately injecting methylprednisolone sodium succinate into the gluteal muscles was used to establish the model, and the treatment group was given intragastric administration of Modified Huoluo Xiaoling Dan (preparation liquid). For the femoral samples of the rats, HE staining was performed and the empty bone lacuna rate was calculated. Three-dimensional reconstruction by Micro-CT was used to analyze and measure the bone histological parameters such as the bone volume fraction (BV/TV), trabecular number (Tb. N), trabecular thickness (Tb. Th) and trabecular separation (Tb. Sp) in the measurement area. Immunohistochemistry method was used to detect the expressions of bone tissue repair markers BMP, OPG and Runx2. For the fecal samples, 16S rDNA technology was used for gut microbiota sequencing and ASV analysis. The α-diversity and β-diversity were used to analyze the species diversity of the gut microbiota, and LEfSe analysis was used to detect the changes in the species differences of the gut microbiota at the family level and genus level.

Results

Modified Huoluo Xiaoling Dan could significantly improve the empty bone lacuna rate of the femoral head (P<0.05) and bone histological parameters such as BV/TV, Tb. N, Tb. Th and Tb. Sp (P<0.01) in the rats with SANFH, and significantly up-regulate the expression levels of bone tissue repair markers BMP, OPG and Runx2 (P<0.01). The results of gut microbiota sequencing showed that α-diversity analysis indicated that there were obvious differences in the species diversity and richness of the gut microbiota structures of the rats in the three groups; β-diversity analysis showed that the flora structure of the treatment group was significantly separated from that of the model group, and was similar to that of the blank group. The difference among groups was greater than that within groups, and the difference was obvious. LEfSe analysis has found that at the family level and genus level, the gut microbiota of the rats in the model group showed significant “pro-inflammatory” characteristics, while the gut microbiota of the rats in the treatment group showed “anti-inflammatory-promoting repair” characteristics.

Conclusion

The effect of Modified Huoluo Xiaoling Dan on the repair of bone tissue in the rats with SANFH is closely related to its regulation of the gut microbiota. The intervention drug effectively reshapes the intestinal microecology disturbed by hormones, further confirming that the reshaping of the flora has a bidirectional regulatory effect on the “gut-osteoporosis” axis, playing a role in preventing and treating SANFH.

Key words

steroid-induced avascular necrosis of the femoral head / Modified Huoluo Xiaoling Dan (加味活络效灵丹) / gut microbiota / bone tissue repair / “gut-osteoporosis” axis

引用本文

张江, 胡宇晴, 张诗宇, 董野, 侯德才. 基于16S rDNA测序探讨加味活络效灵丹对激素性股骨头坏死大鼠“肠-骨”轴的影响. 中华中医药学刊, 2026 , 44 (9) : 53 -58 . DOI: 10.13193/j.issn.1673-7717.2026.09.010

Jiang ZHANG, Yuqing HU, Shiyu ZHANG, Ye DONG, Decai HOU. Effect of Modified Huoluo Xiaoling Dan (加味活络效灵丹) on “Gut-osteoporosis” Axis of Rats with Hormone Induced Femoral Head Necrosis Based on 16S rDNA Sequencing[J]. Chinese Archives of Traditional Chinese Medicine, 2026 , 44 (9) : 53 -58 . DOI: 10.13193/j.issn.1673-7717.2026.09.010

基金

收起

国家自然科学基金项目(82305275)
辽宁中医药大学中医脏象理论及应用教育部重点实验室开放基金项目(zyzx2007)
沈阳市第一批中医药专家学术经验继承项目(20221205)
辽宁中医药大学教学改革研究项目(LNZYJG2024027)
辽宁中医药大学大学生创新创业训练计划项目(X202310162078)

参考文献

收起

文献

收起

参考文献引证文献

排序方式：

[1]

肖涟波, 梁倩倩. 股骨头坏死中西医结合诊疗专家共识[J]. 世界中医药, 2023, 18(7): 901-910.

[2]

中国中医药研究促进会中医骨伤临床循证医学分会, 何伟, 王智勇. 非创伤性股骨头坏死中西医结合诊疗专家共识[J]. 中医正骨, 2024, 36(9): 1-11.

[3]

徐芳茹, 周全. 基于“肠-关节轴”浅析“从脾治痹”[J]. 安徽中医药大学学报, 2022, 41(3): 1-5.

[4]

侯伟. 基于“治痿独取阳明”理论探究激素性股骨头坏死患者肠道菌群分布规律的临床研究[D]. 南京: 南京中医药大学, 2022.

[5]

翁汭, 何晓铭, 何敏聪, 等. 基于络病理论探讨股骨头坏死患者的疼痛特点及疼痛区域经脉循行特点[J]. 中医正骨, 2023, 35(12): 25-29.

[6]

乌日莎娜, 沈莹姗, 袁颖嘉, 等. 基于“瘀去、新生、骨合”理论从细胞层面探讨祛瘀、活血及补肾中药治疗股骨头坏死的作用机制[J]. 中医正骨, 2022, 34(4): 61-63, 69.

[7]

贾振华. 肺疫证治规律探讨与连花清瘟组方新解[J]. 中国实验方剂学杂志, 2024, 30(17): 203-210.

[8]

王丽丽, 李群, 胡智星, 等. 基于氧化脂质代谢组和转录组学整合策略探索健脾活骨方治疗大鼠激素性股骨头坏死的作用机制[J]. 中国实验方剂学杂志, 2025, 31(11): 190-199.

[9]

席洪钟, 陈浩, 何帅, 等. 右归饮通过调节PPARγ通路减轻骨髓间充质干细胞成脂分化治疗糖皮质激素诱导的骨坏死[J/OL]. 中国中药杂志, 1-14[2025-04-06]. https://doi.org/10.19540/j.cnki.cjcmm.20250307.401.

[10]

李金宗, 侯德才, 巩雪. 牛膝-续断治疗股骨头坏死的作用机制[J]. 中医药临床杂志, 2022, 34(7): 1294-1301.

[11]

李伟举, 陈晓聪, 曾家莹, 等. 参骨颗粒通过肠道菌群改善骨质疏松的研究[J]. 中国骨质疏松杂志, 2024, 30(9): 1341-1347, 1352.

[12]

林青霞, 聂春梅, 车润莉, 等. 基于代谢组学和肠道菌群探讨正清风痛宁缓释片治疗膝骨关节炎作用机制[J]. 中国中药杂志, 2024, 49(23): 6417-6428.

[13]

吴霜, 袁立霞, 廖晴, 等. 基于16S rDNA测序研究当归拈痛汤对膝骨关节炎小鼠肠道菌群的影响[J]. 中国实验方剂学杂志, 2024, 30(11): 9-17.

[14]

薛鹏, 杜斌, 刘锌, 等. 基于肠-骨轴探讨肠道菌群对骨代谢影响的研究进展[J]. 中国骨质疏松杂志, 2023, 29(1): 84-88.

[15]

巩彦龙, 黄九妹, 张杰, 等. 肠道屏障与骨代谢[J]. 中国骨质疏松杂志, 2024, 30(12): 1815-1819.

[16]

韦雨柔, 田佳庆, 肖方骏, 等. 活血通络胶囊对激素性股骨头坏死大鼠肠道菌群影响的研究[J]. 中国全科医学, 2023, 26(29): 3674-3682.

[17]

XIAO W P, SU J B, GAO X J, et al. The microbiota-gut-brain axis participates in chronic cerebral hypoperfusion by disrupting the metabolism of short-chain fatty acids[J]. Microbiome, 2022, 10(1): 62.

[18]

邰希尧. 活络效灵丹对激素诱导的股骨头坏死大鼠骨修复的影响[D]. 沈阳: 辽宁中医药大学, 2024.

[19]

MA T C, WANG Y, MA J X, et al. Research progress in the pathogenesis of hormone-induced femoral head necrosis based on microvessels: a systematic review[J]. J Orthop Surg Res, 2024, 19(1): 265.

[20]

CHEN D, ZHONG D M, MEI R H, et al. Screening and identification of potential key biomarkers for glucocorticoid-induced osteonecrosis of the femoral head[J]. J Orthop Surg Res, 2023, 18(1): 28.

[21]

何帅, 刘锌, 杜斌, 等. 肠道菌群变化对大鼠激素性股骨头坏死造模效果的影响[J]. 解放军医学杂志, 2023, 48(5): 552-559.

[22]

杨祥军, 王丛悦, 徐西林, 等. 虎杖苷调节JAK2/STAT3信号通路治疗大鼠激素性股骨头坏死的实验研究[J]. 中国骨伤, 2025, 38(2): 195-203.

[23]

李文茜, 田亮玉, 张瑾, 等. 基于WGCNA和动物实验探究骨坏死康复丸治疗激素性股骨头坏死的作用机制[J]. 中药新药与临床药理, 2024, 35(9): 1306-1318.

[24]

申意伟, 张晓峰, 李雪, 等. 活骨灌注液干预骨坏死的病理形态学、血管新生和成骨分化的实验研究[J]. 时珍国医国药, 2022, 33(4): 831-834.

[25]

TELLO RUBIO B, BUGAULT F, BAUDON B, et al. Molecular mechanisms underpinning the circulation and cellular uptake of Mycobacterium ulcerans toxin mycolactone[J]. Front Pharmacol, 2021, 12: 733496.

[26]

CHENG H Y, GUAN X, CHEN D K, et al. The Th17/treg cell balance: a gut microbiota-modulated story[J]. Microorganisms, 2019, 7(12): 583.

[27]

WANG Y C, XIAO J, WEI S M, et al. Protective effect of zinc gluconate on intestinal mucosal barrier injury in antibiotics and LPS-induced mice[J]. Front Microbiol, 2024, 15: 1407091.

[28]

SHAO W K, WANG P, LV X, et al. Unraveling the role of endothelial dysfunction in osteonecrosis of the femoral head: a pathway to new therapies[J]. Biomedicines, 2024, 12(3): 302-312.

[29]

WANG S S, WANG S J, WANG X N, et al. Effects of icariin on modulating gut microbiota and regulating metabolite alterations to prevent bone loss in ovariectomized rat model[J]. Front Endocrinol, 2022, 13: 874849.

[30]

ZANG R X, ZHOU R, LI Y D, et al. The probiotic Lactobacillus plantarum alleviates colitis by modulating gut microflora to activate PPARγ and inhibit MAPKs/NF-κB[J]. Eur J Nutr, 2024, 64(1): 32.

[31]

HUANG H, HUANG J, HUANG W D, et al. Breast milk jaundice affects breastfeeding: From the perspective of intestinal flora and SCFAs-GPR41/43[J]. Front Nutr, 2023, 10: 1121213.

[32]

YING M X, YU Q, ZHENG B, et al. Cultured Cordyceps sinensis polysaccharides modulate intestinal mucosal immunity and gut microbiota in cyclophosphamide-treated mice[J]. Carbohydr Polym, 2020, 235: 115957.

[33]

魏秋实. 股骨头坏死态靶辨治体系的构建[J]. 中医正骨, 2024(9): 15-18.

[34]

景田园, 王平, 王仪, 等. 鹿仲健骨汤治疗股骨头坏死模型大鼠: 肠道菌群及血清激素变化[J]. 中国组织工程研究, 2024, 28(35): 5598-5605.

2026年第44卷第9期

PDF下载

引用本文

BibTeX

文章信息

doi: 10.13193/j.issn.1673-7717.2026.09.010

首发时间：2026-10-20
出版时间：2026-09-10

补充材料

相关文章

文章信息

作者

出版历史

基金

国家自然科学基金项目(82305275)

辽宁中医药大学中医脏象理论及应用教育部重点实验室开放基金项目(zyzx2007)

沈阳市第一批中医药专家学术经验继承项目(20221205)

辽宁中医药大学教学改革研究项目(LNZYJG2024027)

辽宁中医药大学大学生创新创业训练计划项目(X202310162078)

作者信息

^1.辽宁中医药大学，辽宁　沈阳　110847

^2.辽宁中医药大学附属医院，辽宁　沈阳　110032

^3.辽宁中医药大学中医脏象理论及应用教育部重点实验室，辽宁　沈阳　110847

通讯作者:

侯德才（1961-），男，辽宁沈阳人，教授、主任医师，博士研究生导师，博士，研究方向：中医药防治股骨头坏死。E-mail：lnzyhdc@163.com。

参考文献

分享链接

https://castjournals.cast.org.cn/joweb/zhzyyxk/CN/10.13193/j.issn.1673-7717.2026.09.010

分享至

全文二维码

扫描看全文

引用本文

BibTeX

本文的引用情况

2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

关闭全屏

BibTeX
EndNote
RefWorks
TxT