Study on the Predictive Value of Cystatin A for Osteonecrosis of the Femoral Head

doi:10.12114/j.issn.1007-9572.2024.0299

Chinese General Practice ›› 2025, Vol. 28 ›› Issue (35): 4464-4472.DOI: 10.12114/j.issn.1007-9572.2024.0299

• Original Research • Previous Articles Next Articles

Study on the Predictive Value of Cystatin A for Osteonecrosis of the Femoral Head

1. Guangzhou University of Chinese Medicine, Guangzhou 510006, China
2. Joint Center, Guangdong Provincial Institute of Traditional Chinese Medicine Orthopedics and Traumatology, Guangzhou 510006, China
3. Orthopedics Department, Doctor Workstation, Jiexi County Hospital of Traditional Chinese Medicine, Jieyang 515400, China
4. Orthopedics Department, State Key Laboratory of Traditional Chinese Medicine Syndrome/Joint Center, Third Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510006, China

Received:2024-07-15 Revised:2024-08-03 Published:2025-12-15 Online:2025-10-15
Contact: WEI Qiushi

胱抑素A对股骨头坏死的预测价值研究

1．510006　广东省广州市，广州中医药大学
2．510006　广东省广州市，广东省中医骨伤研究院关节中心
3．515400　广东省揭阳市揭西县中医医院博士工作站骨科
4．510006　广东省广州市，中医证候全国重点实验室/骨科　广州中医药大学第三附属医院关节中心

通讯作者: 魏秋实
作者简介:
作者贡献：
田佳庆提出主要研究目标，负责研究的构思与设计，研究的实施，撰写论文；彭鹏、韦雨柔、肖方骏、何宪顺进行数据的收集与整理，统计学处理，图、表的绘制与展示；林锟、陆舜负责临床标本的收集与整理；何敏聪、刘良燕、魏秋实进行论文的修订；魏秋实负责文章的质量控制与审查，对文章整体负责，监督管理。
基金资助:
国家自然科学基金资助项目(82274544，82004392); 广东省基础与应用基础基金资助项目(2023A1515010551); 广东省中医骨伤研究院开放课题重点项目(GYH202101-01，GYH202101-04)

Abstract

Abstract:

Background

Osteonecrosis of the femoral head (ONFH) is a clinically refractory orthopedic disease with a relatively high incidence and poor prognosis. Currently, magnetic resonance imaging (MRI) is the "gold standard" for diagnosing ONFH, but its examination process is cumbersome and the cost is high, so it cannot be used as a tool for routine screening. Therefore, it is urgent to develop new molecular biomarkers for the diagnosis of ONFH.

Objective

To study the serum cystatin A (CSTA) level in ONFH patients and evaluate its diagnostic value as a biomarker for ONFH.

Methods

Twenty-one ONFH patients who visited the Third Affiliated Hospital of Guangzhou University of Chinese Medicine from May to November 2023 were selected as the experimental group, and 33 healthy volunteers were recruited as the control group at the same time. Serum and femoral head tissue samples were collected from the subjects. Among them, the femoral head tissue samples included 17 patients with ONFH from the experimental group and 6 patients who underwent surgery for femoral neck fractures during the same period, who were designated as the femoral neck fracture (FNF) group. The serum CSTA level was detected by ELISA; The bone tissue morphology was observed by HE staining, and the bone microstructure was detected by CT; Trap staining was used to evaluate the osteoclast differentiation ability in different bone tissues; The expression level of CSTA in bone tissues was evaluated by immunohistochemistry, immunofluorescence staining, and Western Blotting; Spearman rank correlation analysis was used to analyze the correlation between CSTA level and ARCO stage; The receiver operating characteristic (ROC) curve was drawn to evaluate the diagnostic value of CSTA.

Results

The results of serum CSTA detection showed that the serum CSTA levels of patients with ARCO stageⅡ-Ⅳ in the experimental group were higher than those in the control group (P<0.05). ARCO stage Ⅱ was lower than ARCO stage Ⅲ, and ARCO stage Ⅲ was lower than ARCO stage Ⅳ (P<0.05). The results of HE staining showed that there were empty bone depressions in ARCO stages Ⅱ, Ⅲ and Ⅳ. The litter rate was higher than that of the FNF group, and that of ARCO stage Ⅲ andⅣ was higher than that of ARCO stage Ⅱ (P<0.05) ; CT quantitative analysis results showed that the bone volume fraction (BV/TV), trabecular bone thickness (Tb.Th), and trabecular bone number (Tb.N) in ARCO Ⅱ and Ⅲ stages were lower than those in the FNF group, and the trabecular bone separation (Tb.Sp) was higher than that in the FNF group (P<0.05) ; Trap staining results showed that the area of Trap-positive cell regions in ARCO Ⅱ and Ⅲ groups was higher than that in the FNF group, and ARCO Ⅱ was higher than ARCO Ⅳ (P<0.05) ; The results of CSTA detection in bone tissues showed that the CSTA expression in ARCO Ⅱ, Ⅲ, and Ⅳ stages was higher than that in the FNF group, and ARCO Ⅳ stage was higher than ARCO Ⅱstage (P<0.05) ; The area under the ROC curve (AUC) of serum CSTA level in diagnosing ONFH was 0.939, with a sensitivity of 78.79% and a specificity of 95.24%.

Conclusion

The CSTA protein can affect the repair of necrotic bone in the femoral head by regulating the formation of osteoclasts. In patients with osteonecrosis of the femoral head, the increased expression of CSTA is positively correlated with the severity of the disease.

Key words: Osteonecrosis of the femoral head, CSTA, Osteoclast, Biomarker, ARCO staging

摘要：

背景

股骨头坏死（ONFH）是一种临床难治性骨科疾病，发病率较高，预后差。目前磁共振成像（MRI）是诊断ONFH的"金指标"，但其检查流程烦琐，费用较高，无法作为常规筛查的工具。因此，亟待研发新型分子生物标志物用于ONFH的诊断。

目的

研究ONFH患者血清胱抑素A（CSTA）水平，评估其作为ONFH生物标志物的诊断价值。

方法

选取2023年5—11月于广州中医药大学第三附属医院就诊的21例ONFH患者为试验组，同期招募33例健康志愿者为对照组。收集受试者血清和股骨头组织样本，其中股骨头组织样本包括试验组17例ONFH患者和同期因股骨颈骨折手术的6例患者（FNF组），采用酶联免疫吸附法（ELISA）检查血清CSTA水平；采用苏木素-伊红（HE）染色观察骨组织形态，并使用CT扫描检测骨微结构；采用Trap染色评估不同骨组织中破骨细胞分化能力；通过免疫组织化学、免疫荧光染色和Western Blotting评估骨组织CSTA的表达水平；采用Spearman秩相关分析探究CSTA水平与国际骨循环研究协会（ARCO）分期之间的相关性；绘制受试者工作特征（ROC）曲线评估CSTA对ONFH的预测价值。

结果

血清CSTA检测结果显示，试验组ARCO Ⅱ ~ Ⅳ期患者血清CSTA水平高于对照组，ARCO Ⅱ期低于ARCO Ⅲ期，ARCO Ⅲ期低于ARCO Ⅳ期（P<0.05）；HE染色结果显示，ARCO Ⅱ、Ⅲ、Ⅳ期空骨陷窝率高于FNF组，ARCO Ⅲ、Ⅳ期高于ARCO Ⅱ期（P<0.05）；CT定量分析结果显示，ARCOⅡ、Ⅲ期骨体积分数（BV/TV）、骨小梁厚度（Tb.Th）、骨梁数量（Tb.N）低于FNF组，骨小梁分离度（Tb.Sp）高于FNF组（P<0.05）；Trap染色结果显示：ARCOⅡ、Ⅲ期Trap阳性细胞区域面积高于FNF组，且ARCOⅡ期高于ARCO Ⅳ期（P<0.05）；骨组织CSTA检测结果显示，ARCO Ⅱ、Ⅲ和Ⅳ期CSTA表达高于FNF组，且ARCO Ⅳ期高于ARCO Ⅱ期（P<0.05）；血清CSTA水平预测ONFH的ROC曲线下面积（AUC）为0.939 （P<0.001），灵敏度为78.79%，特异度为95.24%。

结论

CSTA蛋白可通过调控破骨细胞的形成来影响股骨头内坏死骨的修复，在股骨头坏死患者中，CSTA表达升高与疾病的严重程度呈正相关。

关键词: 股骨头坏死, CSTA, 破骨细胞, 生物标志物, ARCO分期

CLC Number:

R 68

TIAN Jiaqing,PENG Peng,WEI Yurou, et al. Study on the Predictive Value of Cystatin A for Osteonecrosis of the Femoral Head[J]. Chinese General Practice, 2025, 28(35): 4464-4472. DOI: 10.12114/j.issn.1007-9572.2024.0299.
田佳庆,彭鹏,韦雨柔等. 胱抑素A对股骨头坏死的预测价值研究[J]. 中国全科医学, 2025, 28(35): 4464-4472. DOI: 10.12114/j.issn.1007-9572.2024.0299.

Figures/Tables 10

References 23

[1]	李志刚，胡海，王春龙，等. 三七活骨丸联合体外冲击波治疗老年早期股骨头缺血性坏死的临床价值[J]. 中国老年学杂志，2024，44（21）：5201-5205.
[2]	Microsurgery Department of the Orthopedics Branch of the Chinese Medical Doctor Association；Group from the Osteonecrosis and Bone Defect Branch of the Chinese Association of Reparative and Reconstructive Surgery；Microsurgery and Reconstructive Surgery Group of the Orthopedics Branch of the Chinese Medical Association.Chinese guideline for the diagnosis and treatment of osteonecrosis of the femoral head in adults[J]. Orthop Surg，2017，9（1）：3-12. DOI：10.1111/os.12302.
[3]	CHAI J L, LU B W, DU H T, et al. Pyroptosis-related potential diagnostic biomarkers in steroid-induced osteonecrosis of the femoral head[J]. BMC Musculoskelet Disord，2023，24（1）：609. DOI：10.1186/s12891-023-06729-8.
[4]	SHAMSI A, BANO B. Journey of cystatins from being mere thiol protease inhibitors to at heart of many pathological conditions[J]. Int J Biol Macromol，2017，102：674-693. DOI：10.1016/j.ijbiomac.2017.04.071.
[5]	杨云博，王钰璞，李行，等. 半胱氨酸酶抑制剂A在口腔鳞癌中的表达及临床意义[J]. 中国口腔颌面外科杂志，2021，19（6）：494-499. DOI：10.19438/j.cjoms.2021.06.003.
[6]	李岳骏. 成骨细胞源性cystatin A在骨质疏松发病中的作用与机制研究[D]. 广州：南方医科大学，2019.
[7]	HE M C, ZHANG J, CHEN X J, et al. Osteoclastic activity was associated with the development of steroid-induced osteonecrosis of femoral head[J]. Artif Cells Nanomed Biotechnol，2020，48（1）：1036-1046. DOI：10.1080/21691401.2020.1774596.
[8]	HINES J T, JO W L, CUI Q J, et al. Osteonecrosis of the femoral head：an updated review of ARCO on pathogenesis，staging and treatment[J]. J Korean Med Sci，2021，36（24）：e177. DOI：10.3346/jkms.2021.36.e177.
[9]	中国中医药研究促进会中医骨伤临床循证医学分会. 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. DOI：10.3969/j.issn.1001-6015.2024.09.001.
[10]	田佳庆，何敏聪，韦雨柔，等. 激素性股骨头坏死囊性变分布规律及病理特点[J]. 中国组织工程研究，2023，27（31）：4996-5001.
[11]	周玉刚，魏歆，李舜. CT联合MRI对股骨头坏死的诊断价值探究[J]. 中国实用医药，2024，19（17）：81-83. DOI：10.14163/j.cnki.11-5547/r.2024.17.020.
[12]	杨晓强，肖欢，田佳庆，等. 活血通络胶囊对激素性股骨头坏死囊性变成血管修复的影响[J]. 中医正骨，2024，36（9）：40-48. DOI：10.3969/j.issn.1001-6015.2024.09.007.
[13]	田佳庆，刘良燕，彭鹏，等. 基于"成骨-成血管"理论探讨全身振动疗法治疗激素性股骨头坏死的效果及作用机制[J]. 中医正骨，2024，36（9）：59-68，82. DOI：10.3969/j.issn.1001-6015.2024.09.009.
[14]	魏秋实. 股骨头坏死态靶辨治体系的构建[J]. 中医正骨，2024，36（9）：15-18. DOI：10.3969/j.issn.1001-6015.2024.09.003.
[15]	LIANG W Q, CHEN Q J, CHENG S S, et al. Skin chronological aging drives age-related bone loss via secretion of cystatin-A[J]. Nat Aging，2022，2（10）：906-922. DOI：10.1038/s43587-022-00285-x.
[16]	MA M, CHEN X F, LU L Y, et al. Identification of crucial genes related to postmenopausal osteoporosis using gene expression profiling[J]. Aging Clin Exp Res，2016，28（6）：1067-1074. DOI：10.1007/s40520-015-0509-y.
[17]	李岳骏. 成骨细胞源性cystatin A在骨质疏松发病中的作用与机制研究[D]. 广州：南方医科大学，2019.
[18]	LIU Z J, ZHENG L S, LI C Z, et al. Correlated with better prognosis，CSTA inhibits metastasis of nasopharyngeal carcinoma cells via suppressing AKT signaling through promoting METTL3 degradation[J]. Biochim Biophys Acta Mol Basis Dis，2023，1869（5）：166696. DOI：10.1016/j.bbadis.2023.166696.
[19]	GAO Y, ZHANG M, ZHANG R, et al. Whole grain brown rice extrudate ameliorates the symptoms of diabetes by activating the IRS1/PI3K/AKT insulin pathway in db/db mice[J]. J Agric Food Chem，2019，67（42）：11657-11664.
[20]	ALZAHRANI A S. PI3K/Akt/mTOR inhibitors in cancer：at the bench and bedside[J]. Semin Cancer Biol，2019，59：125-132.
[21]	区作明，梁伟春，李兰. 养血固肾汤通过刺激骨髓间质干细胞对破骨细胞形成的调控及对IGF1R-PI3K-AKT信号通路的作用[J]. 中国老年学杂志，2023，43（9）：2244-2247. DOI：10.3969/j.issn.1005-9202.2023.09.051.
[22]	XIAO D L, ZHOU Q, GAO Y B, et al. PDK1 is important lipid kinase for RANKL-induced osteoclast formation and function via the regulation of the Akt-GSK3β-NFATc1 signaling cascade[J]. J Cell Biochem，2020，121（11）：4542-4557. DOI：10.1002/jcb.29677.
[23]	LU J W, KUANG Z H, CHEN T, et al. Isoalantolactone inhibits RANKL-induced osteoclast formation via multiple signaling pathways[J]. Int Immunopharmacol，2020，84：106550. DOI：10.1016/j.intimp.2020.106550.

组别	例数	空骨陷窝率（%）	骨小梁参数
组别	例数	空骨陷窝率（%）	BV/TV	Tb.N	Tb.Sp	Tb.Th
FNF组	6	5.36±1.77	71.21±2.34	1.71±0.03	0.48±0.05	0.43±0.04
ARCO Ⅱ期	6	25.38±2.78^a	41.53±2.42^a	1.04±0.14^a	0.71±0.03^a	0.38±0.02^a
ARCO Ⅲ期	5	76.26±3.35^ab	25.05±2.54^ab	0.87±0.03^a	1.11±0.07^ab	0.24±0.03^ab
ARCO Ⅳ期	6	85.40±4.45^ab	43.54±2.53^a	1.34±0.13	0.60±0.03^ac	0.38±0.04
F值		705.318	302.842	68.964	178.943	36.294
P值		<0.001	<0.001	<0.001	<0.001	<0.001

组别	例数	空骨陷窝率（%）	骨小梁参数
组别	例数	空骨陷窝率（%）	BV/TV	Tb.N	Tb.Sp	Tb.Th
FNF组	6	5.36±1.77	71.21±2.34	1.71±0.03	0.48±0.05	0.43±0.04
ARCO Ⅱ期	6	25.38±2.78^a	41.53±2.42^a	1.04±0.14^a	0.71±0.03^a	0.38±0.02^a
ARCO Ⅲ期	5	76.26±3.35^ab	25.05±2.54^ab	0.87±0.03^a	1.11±0.07^ab	0.24±0.03^ab
ARCO Ⅳ期	6	85.40±4.45^ab	43.54±2.53^a	1.34±0.13	0.60±0.03^ac	0.38±0.04
F值		705.318	302.842	68.964	178.943	36.294
P值		<0.001	<0.001	<0.001	<0.001	<0.001

组别	例数	Trap染色结果	CSTA
组别	例数	Trap染色结果	免疫组织化学染色	免疫荧光染色	Western Blotting
FNF组	6	0.18±0.04	0.04±0.03	0.04±0.01	0.36±0.02
ARCO Ⅱ期	6	1.00±0.10^a	0.40±0.15	0.20±0.06	0.37±0.01
ARCO Ⅲ期	5	0.60±0.10^a	0.67±0.15^ab	0.65±0.17^ab	0.67±0.05^ab
ARCO Ⅳ期	6	0.32±0.05	1.00±0.36^abc	1.00±0.24^abc	1.00±0.03^abc
F值		52.394	33.348	50.027	324.013
P值		<0.001	<0.001	<0.001	<0.001

组别	例数	Trap染色结果	CSTA
组别	例数	Trap染色结果	免疫组织化学染色	免疫荧光染色	Western Blotting
FNF组	6	0.18±0.04	0.04±0.03	0.04±0.01	0.36±0.02
ARCO Ⅱ期	6	1.00±0.10^a	0.40±0.15	0.20±0.06	0.37±0.01
ARCO Ⅲ期	5	0.60±0.10^a	0.67±0.15^ab	0.65±0.17^ab	0.67±0.05^ab
ARCO Ⅳ期	6	0.32±0.05	1.00±0.36^abc	1.00±0.24^abc	1.00±0.03^abc
F值		52.394	33.348	50.027	324.013
P值		<0.001	<0.001	<0.001	<0.001

Study on the Predictive Value of Cystatin A for Osteonecrosis of the Femoral Head

胱抑素A对股骨头坏死的预测价值研究

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 10

References 23

Related Articles 15

Recommended Articles

Metrics

Comments

组别	例数	CSTA水平
对照组	21	12.47±5.38
ARCO Ⅱ期	10	19.96±6.43^a
ARCO Ⅲ期	13	27.26±4.21^ab
ARCO Ⅳ期	10	33.76±5.55^abc
F值		233.374
P值		<0.001

[1]	LI Ling, LI Yaping, QIAN Shixing, NIE Jing, LU Chunhua, LI Xia. Research on Influencing Factors and Risk Prediction of Cognitive Function in Community-dwelling Middle-aged and Elderly People [J]. Chinese General Practice, 2025, 28(30): 3773-3778.
[2]	NONG Jingtang, YANG Chengmin, MO Shenglong, LU Zhicheng, TANG Lina, JIAN Chongdong, SHANG Jingwei. Research Progress on Prognostic Biomarkers after Cerebral Infarction: Mechanisms and Clinical Applications [J]. Chinese General Practice, 2025, 28(29): 3711-3720.
[3]	LI Jiaxin, LIU Zhonghui, XIE Shuo, FU Zhifang, SUN Dan, JIAO Hongmei. Trajectory in Biomarkers of Metabolic and Inflammatory States as Early Predictors of Chronic Critical Illness in Aging Patients [J]. Chinese General Practice, 2025, 28(24): 2993-2999.
[4]	MA Xiaoxuan, WANG Rongqi. Research Progress in Metabolomics for HCC Diagnosis and Treatment [J]. Chinese General Practice, 2025, 28(08): 1017-1022.
[5]	MAIERHABA· Maimaitiaili, ZHANG Kainan, ZHAO Hui, YAKUFU· Tuoheti, YE Jianwei, LYU Guodong. Investigating Serum PARP2 as a Potential Diagnostic Biomarker for Hepatocellular Carcinoma [J]. Chinese General Practice, 2024, 27(32): 4060-4065.
[6]	PENG Yilun, LI Yang, WANG Xiaotao. Study on the Mechanisms of Multiple Myeloma Cells Promoting M2 Macrophage Polarization through PI3K/AKT Signaling Pathway [J]. Chinese General Practice, 2024, 27(08): 978-984.
[7]	YUAN Dehui, LI Yuhong, XIONG Min, YU Min, MA Ruiliang, YANG Fangfang, YU Qiaozhi, WANG Minghuan. Status and Influencing Factors of Allostatic Load in Pregnant Women at Different Trimesters [J]. Chinese General Practice, 2023, 26(32): 4057-4063.
[8]	LI Yiru, LI Yuhong. Progress in Methods for the Assessment of Allostatic Load and Its Impact on Health [J]. Chinese General Practice, 2023, 26(32): 4051-4056.
[9]	WEI Yurou, TIAN Jiaqing, XIAO Fangjun, HE Xianshun, ZHAN Zhiwei, WEI Tengfei, LIN Tianye, HE Mincong, WEI Qiushi. Effect of Huoxue Tongluo Capsule on Intestinal Flora of Rats with Steroid-induced Osteonecrosis of the Femoral Head [J]. Chinese General Practice, 2023, 26(29): 3674-3682.
[10]	JI Mengying, LI Yujing, CHEN Xing, DAI Huihua, SUN Ying. Prediction Value of B-ultrasound with Tumor Markers for Malignant Transformation of Mucinous Ovarian Tumors [J]. Chinese General Practice, 2023, 26(24): 3022-3027.
[11]	XU Ting, JI Minghui, CHEN Yimeng, GAO Yu, ZHU Hanfei, DING Lingyu, XU Qin. Advances in Multiomic Analyses of Frailty Biomarkers in the Elderly [J]. Chinese General Practice, 2023, 26(23): 2871-2876.
[12]	GOU Dengqun, ZHANG Lu, XU Yuanli, JIANG Mingjiao, WU Hemei, TAO Ming. Neutrophil-to-lymphocyte Ratio and Red Cell Distribution Width as Potential Biomarkers of Frailty: a Scoping Review [J]. Chinese General Practice, 2023, 26(17): 2169-2175.
[13]	XU Lili, HONG Yunzhe, LI Zhihui, YU Ningxia, DI Jiaqi, YANG Shuguang, LIN Qingqing, YU Xueqing. Prognostic Biomarkers in Idiopathic Pulmonary Fibrosis: a Recent Review [J]. Chinese General Practice, 2023, 26(03): 372-379.
[14]	Tuoheti YAKUFU·, Kainan ZHANG, Hui ZHAO, Dawuti WUBULITALIFU·, Rui ZHANG, Renyong LIN, Guodong LYU. Performance of Serum m3AchR-Ab for Alpha-fetoprotein-negative Hepatocellular Carcinoma Diagnosis [J]. Chinese General Practice, 2022, 25(17): 2110-2114.
[15]	Jiehong KANG, Xiangyun CHANG, Yurong ZHU, Tao BA, Yuan SUN. Association of Cyclic RNAs with the Risk of Type 2 Diabetes Mellitus in Han Chinese [J]. Chinese General Practice, 2022, 25(15): 1863-1868.