[1]徐西林 王银仓 任树军 李小冬 尚东雨 高宇鹤 刘博 姜益常.活骨方对酒精性骨质疏松大鼠骨髓微环境相关炎症-氧化失衡及衰老信号的影响[J].中国中医骨伤科杂志,2026,34(05):1-9.[doi:10.20085/j.cnki.issn1005-0205.260501]
　XU Xilin,WANG Yincang,REN Shujun,et al.Effects of Huogu Formula on Bone Marrow Microenvironment-Related Inflammatory-Oxidative Imbalance and Senescence-Related Signaling in Rats with Alcoholic Osteoporosis[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2026,34(05):1-9.[doi:10.20085/j.cnki.issn1005-0205.260501]
点击复制

活骨方对酒精性骨质疏松大鼠骨髓微环境相关炎症-氧化失衡及衰老信号的影响()

《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第34卷
期数:
2026年05期
页码:
1-9
栏目:
基础研究
出版日期:
2026-05-10

文章信息/Info

Title:
Effects of Huogu Formula on Bone Marrow Microenvironment-Related Inflammatory-Oxidative Imbalance and Senescence-Related Signaling in Rats with Alcoholic Osteoporosis
文章编号:
1005-0205(2026)05-0001-09
作者:
徐西林 王银仓 任树军 李小冬 尚东雨 高宇鹤 刘博 姜益常
1黑龙江中医药大学附属第三医院(哈尔滨,150036)
2黑龙江中医药大学附属第一医院
3黑龙江中医药大学附属第二医院
Author(s):
XU Xilin1; WANG Yincang2; REN Shujun2; LI Xiaodong1; SHANG Dongyu3; GAO Yuhe1; LIU Bo1; JIANG Yichang2△
1The Third Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150036,China; 2The First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150040,China; 3The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150006,China.
关键词:
活骨方;  酒精性骨质疏松;  骨髓微环境;  炎症;  氧化应激;  p16/p21/p53蛋白
Keywords:
Huogu formula;  alcoholic osteoporosis;  bone marrow microenvironment;  inflammation;  oxidative stress;  p16/p21/p53 protein
分类号:
R285.5; R274
DOI:
10.20085/j.cnki.issn1005-0205.260501
文献标志码:
A
摘要:
目的:探讨活骨方对酒精性骨质疏松(AOP)大鼠骨髓微环境相关的炎症反应、股骨远端松质骨微结构、骨组织抗氧化防御及p16/p21/p53相关蛋白变化的影响,并结合骨代谢指标与骨髓脂肪沉积变化评价其干预作用。方法:选取6～8周龄,体重(200±20)g的无特定病原体(SPF)级雄性SD大鼠50只,按随机数字表法分为正常组(N组)、酒精性骨质疏松模型组(AOP组)和活骨方低/中/高剂量组(L/M/H组),每组10只。除N组外,其余各组采用20%体积分数酒精10 mL/kg灌胃,每周6次,连续20周建立酒精性骨质疏松模型。于第16周采用股骨远端Micro-CT对模型进行阶段性验证; 自第16周起,在持续酒精干预基础上,L/M/H组分别给予活骨方0.98,1.96,3.92 g/kg灌胃,每周6次,连续4周。第20周末再次行股骨远端Micro-CT观察松质骨微结构,检测血清IL-1β、IL-6、CXCL10、MCP-1及Ca、P、PINP、CTX-1水平; 检测骨组织总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)水平; 采用Western Blot法检测股骨组织p16、p21、p53蛋白表达; 采用茜素红染色和油红O染色观察骨矿化及骨髓脂肪沉积变化。结果:第16周Micro-CT结果显示,与N组比较,AOP组股骨远端松质骨骨小梁明显稀疏、变细,骨小梁组织平均骨密度降低,差异有统计学意义(P<0.01),骨小梁分离度(Tb.Sp)升高,差异有统计学意义(P<0.01),骨小梁厚度(Tb.Th)降低,差异有统计学意义(P<0.01),骨小梁数目(Tb.N)降低,差异有统计学意义(P<0.001),骨体积分数(BV/TV)降低,差异有统计学意义(P<0.000 1),提示模型已成功建立。第20周末,与N组比较,AOP组上述松质骨微结构异常仍然存在。与AOP组比较,活骨方干预后股骨远端松质骨微结构均有不同程度改善,其中高剂量组改善更为明显,表现为骨小梁组织平均骨密度、Tb.Th、Tb.N及BV/TV升高,Tb.Sp降低,差异有统计学意义(P<0.05)。同时,与N组比较,AOP组血清IL-1β、IL-6、CXCL10、MC-1水平升高,Ca、P水平下降,骨组织T-AOC、SOD、CAT水平下降,股骨组织p16、p21、p53蛋白表达上调,差异均有统计学意义(P<0.05),并伴骨髓脂肪沉积增加及骨代谢失衡; 活骨方干预后上述异常均得到不同程度改善,其中高剂量组改善幅度相对较大,差异有统计学意义(P<0.05)。结论:活骨方可改善酒精性骨质疏松大鼠股骨远端松质骨微结构损伤,减轻炎症因子异常升高、骨组织抗氧化能力下降、p16/p21/p53 相关蛋白异常表达、骨髓脂肪沉积增加及骨代谢紊乱,其保护作用可能与改善骨髓微环境相关的异常状态有关。
Abstract:
Objective:To investigate the effects of Huogu formula on bone marrow microenvironment-related inflammatory response,distal femoral trabecular microarchitecture,bone antioxidant defense,and p16/p21/p53-related protein changes in rats with alcoholic osteoporosis(AOP),and to evaluate its intervention effects in combination with bone metabolism indices and bone marrow fat deposition.Methods:Fifty SPF male Sprague-Dawley rats aged 68 weeks and weighing(200±20)g were randomly divided into normal group(N group),AOP model group(AOP group),and low-,medium-,and high-dose Huogu formula groups(L,M,and H groups),with 10 rats in each group.Except for the N group,rats in all groups received intragastric administration of 20%(vol/vol)ethanol at 10 mL/kg,6 times per week for 20 consecutive weeks to establish the AOP model.At week 16,distal femoral Micro-CT was performed for stage validation of the model.From week 16,on the basis of continued ethanol intervention,rats in the L,M,and H groups were given Huogu formula by gavage at doses of 0.98,1.96,and 3.92 g/kg,respectively,6 times per week for 4 consecutive weeks.At week 20,distal femoral trabecular microarchitecture was re-evaluated by Micro-CT.Serum levels of IL-1β,IL-6,CXCL10,MCP-1,Ca,P,PINP,and CTX-1 were measured.Total antioxidant capacity(T-AOC),superoxide dismutase(SOD),and catalase(CAT)levels in bone tissue were determined.Protein expression of p16,p21,and p53 in femoral tissue was detected by Western Blot.Bone mineralization and bone marrow fat deposition were observed by Alizarin Red staining and Oil Red O staining,respectively.Results:At week 16,Micro-CT showed that,compared with the N group,rats in the AOP group exhibited markedly sparse and thinned trabeculae in the distal femur,accompanied by decreased trabecular tissue mean BMD(P<0.01),reduced trabecular thickness(Tb.Th,P<0.01),trabecular number(Tb.N,P<0.001),and bone volume fraction(BV/TV,P<0.000 1),as well as increased trabecular separation(Tb.Sp,P<0.01),indicating successful model establishment.At week 20,these trabecular microarchitectural abnormalities remained in the AOP group.Compared with the AOP group,Huogu formula improved distal femoral trabecular microarchitecture to varying degrees,with more marked improvement in the high-dose group,as evidenced by increased trabecular tissue mean BMD,Tb.Th,Tb.N,and BV/TV and decreased Tb.Sp(all P<0.05).Meanwhile,compared with the N group,the AOP group showed increased serum IL-1β,IL-6,CXCL10,and MCP-1 levels(all P<0.05), accompanied by decreased serum Ca and P leves,decreased bone tissue T-AOC,SOD,and CAT levels(all P<0.05),upregulated p16,p21,and p53 protein expression in femoral tissue(all P<0.05),increased bone marrow fat deposition,and disordered bone metabolism.Huogu formula ameliorated these abnormalities to different extents,with relatively greater improvement in the high-dose group(all P<0.05).Conclusion:Huogu formula can ameliorate distal femoral trabecular microarchitectural damage,abnormal elevation of inflammatory factors,decreased bone antioxidant capacity,abnormal expression of p16/p21/p53-related proteins,increased bone marrow fat deposition,and bone metabolic disorder in AOP rats.Its protective effect may be related to the improvement of abnormalities associated with the bone marrow microenvironment.

参考文献/References:

[1] KE Y M,HU H F,ZHANG J L,et al.Alcohol consumption and risk of fractures:a systematic review and dose-response meta-analysis of prospective cohort studies[J].Advances in Nutrition,2023,14(4):599-611.
[2] GODOS J,GIAMPIERI F,CHISARI E,et al.Alcohol consumption,bone mineral density,and risk of osteoporotic fractures:a dose-response meta-analysis[J].International Journal of Environmental Research and Public Health,2022,19(3):1515.
[3] GUO M,HUANG Y L,WU Q,et al.Chronic ethanol consumption induces osteopenia via activation of osteoblast necroptosis[J].Oxidative Medicine and Cellular Longevity,2021:3027954.
[4] SYED HASHIM S A,NAINA MOHAMED I,MOHAMED N.The effects of acute and chronic alcohol administration and withdrawal on bone microstructure,mechanical strength,and remodeling protein expression and their relation to an antioxidant and FGF23 in vivo[J].Biomedicines,2024,12(7):1515.
[5] LUO J,LI L,SHI W S,et al.Oxidative stress and inflammation:roles in osteoporosis[J].Frontiers in Immunology,2025,16:1611932.
[6] XU J,YU L X,LIU F,et al.The effect of cytokines on osteoblasts and osteoclasts in bone remodeling in osteoporosis:a review[J].Frontiers in Immunology,2023,14:1222129.
[7] IANTOMASI T,ROMAGNOLI C,PALMINI G,et al.Oxidative stress and inflammation in osteoporosis:molecular mechanisms involved and the relationship with microRNAs[J].International Journal of Molecular Sciences,2023,24(4):3772.
[8] MARCUCCI G,DOMAZETOVIC V,NEDIANI C,et al.Oxidative stress and natural antioxidants in osteoporosis:novel preventive and therapeutic approaches[J].Antioxidants,2023,12(2):373.
[9] LAWRENCE M,GOYAL A,PATHAK S,et al.Cellular senescence and inflammaging in the bone:pathways,genetics,anti-aging strategies and interventions[J].International Journal of Molecular Sciences,2024,25(13):7411.
[10] 杨超富,谭国庆,徐展望.骨质疏松中衰老相关分泌表型调控机制的研究进展[J].中国全科医学,2024,27(29):3685-3695.
[11] 王薇,杨钰婷,白金霞,等.细胞衰老及其驱动基因与骨质疏松症研究进展[J].中国骨质疏松杂志,2025,31(2):215-221.
[12] PIGNOLO R J,LAW S F,CHANDRA A.Bone aging,cellular senescence,and osteoporosis[J].JBMR Plus,2021,5(4):e10488.
[13] CHENG X Y,CHEN Y D,ZHOU X C,et al.Immunoporosis:the hidden link between aging immune cells and bone fragility[J].Journal of Orthopaedic Translation,2025,53:325-335.
[14] GUO H Y,BI Y J,ZHANG G Z,et al.Alcohol-induced bone loss driven by dysregulated spatial distribution of gut microbiota and PGD2-IL17 pathway-mediated osteoclast activation[J].Frontiers in Microbiology,2025,16:1551028.
[15] PACCOU J,BADR S,LOMBARDO D,et al.Bone marrow adiposity and fragility fractures in postmenopausal women:the ADIMOS case-control study[J].The Journal of Clinical Endocrinology & Metabolism,2023,108(10):2526-2536.
[16] MARINELLI BUSILACCHI E,MORSIA E,POLONI A.Bone marrow adipose tissue[J].Cells,2024,13(9):724.
[17] MIURA M,KITAURA H,OHORI F,et al.Role of CXCL10 released from osteocytes in response to TNF-α stimulation on osteoclasts[J].Scientific Reports,2025,15:3040.
[18] 郭曹培,程飘涛,杨成兵,等.骨免疫与骨代谢[J].中国组织工程研究,2024,28(14):2261-2266.
[19] 王亮,郭翔云,秦金然,等.两样本孟德尔随机化研究炎症因子与骨质疏松症的因果关联[J].中国骨质疏松杂志,2025,31(1):62-69.
[20] 匡昱林,匡浩铭,李洪芳,等.循环炎症细胞因子与骨质疏松症的双向孟德尔随机化研究[J].中国骨质疏松杂志,2025,31(1):70-77.
[21] WANG L L,WANG S,DAI X,et al.Salvia miltiorrhiza in osteoporosis:a review of its phytochemistry,traditional clinical uses and preclinical studies(2014-2024)[J].Frontiers in Pharmacology,2024,15:1483431.
[22] MA C,MO L,WANG Z Z,et al.Dihydrotanshinone I attenuates estrogen-deficiency bone loss through RANKL-stimulated NF-κB,ERK and NFATc1 signaling pathways[J].International Immunopharmacology,2023,123:110572.
[23] LIN J G,WANG Q Q,ZHOU S M,et al.Tetramethylpyrazine:a review on its mechanisms and functions[J].Biomedicine & Pharmacotherapy,2022,150:113005.
[24] HUANG S W,CHEN J M,LIU X H,et al.Evaluation of the pharmaceutical activities of Chuanxiong,a key medicinal material in traditional Chinese medicine[J].Pharmaceuticals,2024,17(9):1157.
[25] ZHANG F Q,LI Q Y,WU J S,et al.Total flavonoids of drynariae rhizoma improve glucocorticoid-induced osteoporosis of rats:UHPLC-MS-based qualitative analysis,network pharmacology strategy and pharmacodynamic validation[J].Frontiers in Endocrinology,2022,13:920931.
(收稿日期:2026-04-05)

备注/Memo

备注/Memo:
基金项目:黑龙江省自然科学基金联合基金培育项目(JJ2025PL0348)
黑龙江省卫生健康委科研课题(20252121010127)
国家自然科学基金项目(82374496)
△通信作者 E-mail:jiangyichang2008@126.com
更新日期/Last Update: 2026-05-15