马骨续筋胶囊通过SDF-1/CXCR4信号通路促进老年大鼠髋部骨折愈合

doi:10.11958/20241261

摘要/Abstract

摘要：

目的探讨马骨续筋胶囊对老年大鼠髋部骨折愈合及基质细胞衍生因子-1（SDF-1）/CXC型趋化因子受体4（CXCR4）信号通路的影响。方法构建老年大鼠髋部骨折模型，将造模成功大鼠随机分为模型组，马骨续筋胶囊低、高剂量组，马骨续筋胶囊高剂量+通路抑制剂（AMD3100）组，每组12只，另取12只健康大鼠作为对照组。X线片观察大鼠髋部骨折愈合情况并进行Garrett评分；酶联免疫吸附试验（ELISA）检测血清骨折愈合相关炎症因子肿瘤坏死因子-α（TNF-α）和白细胞介素-10（IL-10）水平；CT扫描分析测定骨体积分数（BV/TV）与骨表面积/体积比（BS/BV）；免疫组化检测骨折愈合因子胰岛素样生长因子（IGF）-1、骨形态发生蛋白（BMP）-2表达；Western blot检测SDF-1/CXCR4信号通路相关蛋白表达。结果模型组较对照组Garrett评分、IL-10水平、BV/TV、BS/BV及IGF-1、BMP-2、SDF-1、CXCR4表达降低，TNF-α水平升高（P<0.05）；马骨续筋胶囊低、高剂量组较模型组Garrett评分、IL-10水平、BV/TV、BS/BV及IGF-1、BMP-2、SDF-1、CXCR4表达升高，TNF-α水平降低（P<0.05）；马骨续筋胶囊高剂量+AMD3100组较马骨续筋胶囊高剂量组Garrett评分、IL-10水平、BV/TV、BS/BV及IGF-1、BMP-2、SDF-1、CXCR4表达降低，TNF-α水平升高（P<0.05）。结论马骨续筋胶囊可促进老年大鼠髋部骨折愈合，其作用机制可能与激活SDF-1/CXCR4信号通路有关。

关键词: 髋骨折, 骨折愈合, 趋化因子CXCL12, 受体，CXCR6, 马骨续筋胶囊

Abstract:

Objective To explore the effects of Magu Xujin capsule on hip fracture healing and stromal cell-derived factor-1/CXC chemokine receptor 4 (SDF-1/CXCR4) signaling pathway in elderly hip fracture rats. Methods The hip fracture model of aged rats was established, and successfully modeled rats were randomly separated into the model group, the low-dose Magu Xujin capsule group, the high-dose Magu Xujin capsule group, the high-dose Magu Xujin capsule+pathway inhibitor group (the high-dose Magu Xujin capsule and AMD3100 group), with 12 rats in each group. An additional 12 healthy rats were selected as the control group. X-ray was used to observe the healing of hip fractures in rats, and Garrett score was performed. Enzyme-linked immunosorbent assay (ELISA) was applied to detect serum levels of inflammatory factors related to fracture healing, such as tumor necrosis factor-alpha (TNF-α) and interleukin-10 (IL-10). CT scanning analysis was used to determine bone volume fraction (BV/TV) and bone surface area/volume ratio (BS/BV). Immunohistochemistry was applied to detect the expression of fracture healing factor insulin-like growth factor-1 (IGF-1) and bone morphogenetic protein-2 (BMP-2). Western blot assay was applied to detect the expression of SDF-1/CXCR4 signaling pathway related proteins. Results Garrett score, IL-10 level, BV/TV, BS/BV, the IGF-1, BMP-2, SDF-1 and CXCR4 expression were lower in the model group than those in the control group, while TNF-α level was higher (P<0.05). Garrett score, IL-10 level, BV/TV, BS/BV, IGF-1, BMP-2, SDF-1 and CXCR4 expression were higher in the low-dose and high-dose Magu Xujin capsule groups than those in the model group, while TNF-α level was lower (P<0.05). Garrett score, IL-10 level, BV/TV, BS/BV, IGF-1, BMP-2, SDF-1 and CXCR4 expression were lower in the high-dose Magu Xujin capsule+AMD3100 group than those in the high-dose Magu Xujin capsule group, while TNF-α level was higher (P<0.05). Conclusion Magu Xujin capsule can promote hip fracture healing in aged rats, and its mechanism may be related to the activation of SDF-1/CXCR4 signaling pathway.

Key words: hip fractures, fracture healing, chemokine CXCL12, receptors, CXCR6, Magu Xujin capsule

中图分类号:

R683

图/表 7

图1 老年大鼠髋部骨折形态

Fig.1 Hip fracture morphology in aged rats

表1 各组大鼠Garrett评分及血清炎症因子水平的比较

Tab.1 Comparison of Garrett score and serum levels of inflammatory factors between the five groups （n=12，$\bar{x}$±s）

组别	Garrett评分/分	TNF-α/（ng/L）	IL-10/（ng/L）
对照组	2.75±0.29	5.22±0.54	33.84±3.49
模型组	0.50±0.06^a	30.56±3.17^a	6.71±0.72^a
马骨续筋胶囊低剂量组	1.50±0.16^b	18.34±1.93^b	18.63±1.95^b
马骨续筋胶囊高剂量组	2.58±0.28^bc	6.75±0.71^bc	31.59±3.36^bc
马骨续筋胶囊高剂量+AMD3100组	1.58±0.17^d	17.48±1.85^d	19.42±2.03^d
F	226.950^＊＊	350.894^＊＊	227.478^＊＊

表2 各组大鼠BV/TV与BS/BV的比较

Tab.2 Comparison of BV/TV and BS/BV of rats between the five groups （n=6，$\bar{x}$±s）

组别	BV/TV	BS/BV
对照组	0.20±0.03	3.28±0.35
模型组	0.05±0.01^a	1.02±0.12^a
马骨续筋胶囊低剂量组	0.12±0.02^b	2.05±0.21^b
马骨续筋胶囊高剂量组	0.19±0.03^bc	3.14±0.33^bc
马骨续筋胶囊高剂量+AMD3100组	0.13±0.02^d	2.19±0.24^d
F	40.778^＊＊	72.788^＊＊

图2 免疫组化检测IGF-1、BMP-2表达（×400） A—E分别为对照组、模型组、马骨续筋胶囊低剂量组、马骨续筋胶囊高剂量组、马骨续筋胶囊高剂量+AMD3100组。

Fig.2 Immunohistochemical detection of IGF-1 and BMP-2 expression (×400)

表3 Comparison of IGF-1 and BMP-2 expression in hip bone tissue of rats between the five groups （n=6，OD值，$\bar{x}$±s）

Tab.3

组别	IGF-1	BMP-2
对照组	0.71±0.08	0.56±0.06
模型组	0.26±0.03^a	0.11±0.02^a
马骨续筋胶囊低剂量组	0.44±0.05^b	0.30±0.03^b
马骨续筋胶囊高剂量组	0.65±0.07^bc	0.52±0.06^bc
马骨续筋胶囊高剂量+AMD3100组	0.48±0.05^d	0.31±0.04^d
F	55.762^＊＊	99.653^＊＊

图3 Western blot检测SDF-1、CXCR4表达 A：对照组；B：模型组；C：马骨续筋胶囊低剂量组；D：马骨续筋胶囊高剂量组；E：马骨续筋胶囊高剂量+AMD3100组。

Fig.3 Western blot analysis of SDF-1 and CXCR4 expression

表4 各组大鼠髋骨组织SDF-1/CXCR4信号通路相关蛋白表达的比较

Tab.4 Comparison of SDF-1/CXCR4 signaling pathway related protein expression in hip bone tissue of rats between the five groups （n=6，$\bar{x}$±s）

组别	SDF-1	CXCR4
对照组	0.85±0.09	0.79±0.08
模型组	0.37±0.04^a	0.33±0.04^a
马骨续筋胶囊低剂量组	0.58±0.06^b	0.54±0.06^b
马骨续筋胶囊高剂量组	0.82±0.09^bc	0.77±0.08^bc
马骨续筋胶囊高剂量+AMD3100组	0.60±0.06^d	0.56±0.06^d
F	46.356^＊＊	49.681^＊＊

参考文献 19

[1]	MEAGHER E, VARGHESE S, HARTY J A, et al. The decline of hip fracture incidence rates over a 10-year period:a single centre experience[J]. Injury, 2021, 52(7):1807-1812. doi:10.1016/j.injury.2021.04.051.
[2]	张玉, 孙炜, 孙维超, 等. 入院红细胞体积分布宽度与老年髋部骨折术后肺部感染的关系及其预测价值[J]. 天津医药, 2022, 50(6):608-612.
	ZHANG Y, SUN W, SUN W C, et al. The predictive value of red cell distribution width for postoperative lung infection in elderly patients with hip fracture[J]. Tianjin Med J, 2022, 50(6):608-612. doi:10.11958/20212647.
[3]	周宝柱, 刘道阔, 张鹏, 等. 马骨续筋胶囊联合可调背伸外固定支具治疗胸腰椎压缩性骨折临床评价[J]. 中国药业, 2021, 30(22):104-107.
	ZHOU B Z, LIU D K, ZHANG P, et al. Clinical evaluation of Magu Xujin capsule combined with adjustable back extension external fixation brace in the treatment of thoracolumbar compression fractures[J]. China Pharmaceutical Industry, 2021, 30(22):104-107. doi:10.3969/j.issn.1006-4931.2021.22.027.
[4]	YE D, CHEN C, WANG Q, et al. Short-wave enhances mesenchymal stem cell recruitment in fracture healing by increasing HIF-1 in callus[J]. Stem Cell Res Ther, 2020, 11(1):382. doi:10.1186/s13287-020-01888-0.
[5]	周欣, 石维祥, 罗晓中. 茶多酚对髋部骨折大鼠VEGF/VEGFR-2信号通路及炎症因子TNF-α和IL-10含量水平的影响[J]. 临床和实验医学杂志, 2021, 20(17):1812-1815.
	ZHOU X, SHI W X, LUO X Z. Effects of tea polyphenols on VEGF/VEGFR-2 signaling pathway and levels of inflammatory factors TNF-α and IL-10 in hip fracture rats[J]. Journal of Clinical and Experimental Medicine, 2021, 20(17):1812-1815. doi:10.3969/j.issn.1671-4695.2021.17.006.
[6]	周宝柱, 刘道阔, 张鹏, 等. 马骨续筋胶囊联合微创经皮椎弓根螺钉固定术对胸腰椎压缩性骨折愈合的影响[J]. 中国药业, 2022, 31(1):95-98.
	ZHOU B Z, LIU D K, ZHANG P, et al. Effect of Magu Xujin capsule combined with minimally invasive percutaneous pedicle screw fixation on the healing of thoracolumbar compression fractures[J]. China Pharmaceutical Industry, 2022, 31(1):95-98. doi:10.3969/j.issn.1006-4931.2022.01.025.
[7]	唐昊, 陈顺有, 卢晓坤, 等. 小鼠SDF-1/CXCR4介导的AKT与ERK通路抑制对延缓骨折愈合的影响[J]. 中外医学研究, 2020, 18(19):6-9.
	TANG H, CHEN S Y, LU X K, et al. Effect of inhibition of AKT and ERK pathways mediated by SDF-1/CXCR4 on delayed fracture healing in mice[J]. Chinese and Foreign Medical Research, 2020, 18(19):6-9. doi:10.14033/j.cnki.cfmr.2020.19.003.
[8]	侯秀秀, 韩广普, 石国君, 等. 老年股骨粗隆部骨折应用马骨续筋胶囊治疗的效果观察[J]. 药店周刊, 2021, 30(1):181.
	HOU X X, HAN G P, SHI G J, et al. Effect of Magu Xujin capsule on elderly patients with trochanteric fracture of femur[J]. Journal of Pharmacies, 2021, 30(1):181.
[9]	齐冰冰, 李国梁. 马骨续筋胶囊联合中医手法复位孟氏架治疗胫腓骨骨折的临床研究[J]. 河北中医, 2022, 44(4):569-573.
	QI B B, LI G L. Clinical study of Magu Xujin capsule combined with traditional Chinese medicine manual reduction of Mengzi frame in treatment of tibia and fibula fracture[J]. Hebei Traditional Chinese Medicine, 2022, 44(4):569-573. doi:10.3969/j.issn.1002-2619.2022.04.010.
[10]	KUSHIOKA J, CHOW S K, TOYA M, et al. Bone regeneration in inflammation with aging and cell-based immunomodulatory therapy[J]. Inflamm Regen, 2023, 43(1):29. doi:10.1186/s41232-023-00279-1.
[11]	WANG L, ZHANG G. Analysis of the effect of mini-nutrition nursing plus Baduanjin rehabilitation exercise on fracture healing,mobility and nutritional status of elderly patients with vertebral or hip fractures[J]. Biotechnol Genet Eng Rev, 2024, 40(3):2508-2519. doi:10.1080/02648725.2023.2200302.
[12]	TORRES H M, ARNOLD K M, OVIEDO M, et al. Inflammatory processes affecting bone health and repair[J]. Curr Osteoporos Rep, 2023, 21(6):842-853. doi:10.1007/s11914-023-00824-4.
[13]	JANTABOON S, SAKUNRANGSIT N, TOEJING P, et al. Lipopolysaccharide impedes bone repair in fcγriib-deficient mice[J]. Int J Mol Sci, 2023, 24(23):16944-16961. doi: 10.3390/ijms242316944.
[14]	陈月, 苗存良, 安志辉, 等. 芹菜素调节OPG/RANKL/RANK信号通路对创伤性骨折大鼠骨折愈合的影响[J]. 国际检验医学杂志, 2023, 44(12):1446-1451.
	CHEN Y, MIAO C L, AN Z H, et al. Effect of apigenin regulating OPG/RANKL/RANK signaling pathway on fracture healing in rats with traumatic fracture[J]. International Journal of Laboratory Medicine, 2023, 44(12):1446-1451. doi:10.3969/j.issn.1673-4130.2023.12.008.
[15]	CUI K, CHEN Y, ZHONG H, et al. Transplantation of IL-10-overexpressing bone marrow-derived mesenchymal stem cells ameliorates diabetic-induced impaired fracture healing in mice[J]. Cell Mol Bioeng, 2020, 13(2):155-163. doi:10.1007/s12195-019-00608-w.
[16]	MOLLAHOSSEINI M, AHMADIRAD H, GOUJANI R, et al. The association between traumatic brain injury and accelerated fracture healing:a study on the effects of growth factors and cytokines[J]. J Mol Neurosci, 2021, 71(1):162-168. doi:10.1007/s12031-020-01640-6.
[17]	PARK Y, LIN S, BAI Y, et al. Dual delivery of BMP2 and IGF1 through injectable hydrogel promotes cranial bone defect healing[J]. Tissue Eng Part A, 2022, 28(17/18):760-769. doi:10.1089/ten.TEA.2022.0002.
[18]	XUE Y, LI Z, WANG Y, et al. Role of the HIF-1α/SDF-1/CXCR4 signaling axis in accelerated fracture healing after craniocerebral injury[J]. Mol Med Rep, 2020, 22(4):2767-2774. doi:10.3892/mmr.2020.11361.
[19]	ZHANG L, JIN L, GUO J, et al. Chronic intermittent hypobaric hypoxia enhances bone fracture healing[J]. Front Endocrinol(Lausanne), 2020,11:582670. doi:10.3389/fendo.2020.582670.