| 桃红四物汤早期干预对大鼠骨折愈合中间充质干细胞归巢的影响 |
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投稿时间：2021-06-19
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| 期刊信息：《中国骨伤》2022年，第35卷，第4期，第367-374页 |
| DOI：10.12200/j.issn.1003-0034.2022.04.014 |
| 基金项目:国家自然科学基金(编号:81874478) |
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中文摘要:
目的: 观察桃红四物汤对骨折大鼠的早期愈合过程中间充质干细胞(mesenchymal stem cells,MSCs)迁移能力、归巢数目及骨痂内细胞因子表达的影响,探讨桃红四物汤通过调控间充质干细胞归巢加速骨折愈合的作用机制。
方法: 建立右侧股骨干开放性骨折大鼠模型。将5周龄雄性Sprague-Dawley(SD)大鼠32只,体重110~130(120±10) g,采用随机数字表法分为对照组,桃红四物汤低、中、高剂量组。对照组予蒸馏水,其他组予以不同剂量的桃红四物汤。术后第1天灌胃,每天2次,连续5 d。术后21 d,采用微型计算机断层扫描技术(micro-computed tomography,Micro-CT)观察骨体积分数即骨体积(bone volume)/组织体积(tissue volume)(BV/TV)和骨密度(bone mineral density,BMD)。术后5 d,体外培养对照组和药物组大鼠外周血MSCs,采用细胞迁移实验检测其迁移能力。免疫组化(immunohistochemistry,IHC)检测骨痂处MSCs的数目。蛋白芯片技术检测骨痂内细胞因子的表达。
结果: Micro-CT结果显示,高剂量组BV/TV高于中剂量组(P=0.032),中剂量组高于低剂量组(P=0.041),对照组和低剂量组无差异性(P=0.651);此外,对照组BMD和低剂量组无差异(P=0.671),低剂量组低于中剂量组(P=0.018),中剂量组低于高剂量组(P=0.008)。细胞迁移实验表明,桃红四物汤能增强外周血MSCs的迁移能力;IHC检测发现,骨痂内CD45-、CD90+、CD29+间充质干细胞较对照组明显增多;蛋白芯片显示,桃红四物汤能促进骨折大鼠骨痂内CINC-1(2.91倍)、CINC-3(1.59倍)、LIX (1.5倍)、Thymus Chemokine(2.55倍)、VEGF(1.22倍)表达,抑制TIMP-1(2.98倍)表达。
结论: 骨折早期予活血化瘀代表方桃红四物汤能明显加速骨折愈合,其作用机制可能与增强外周血MSCs迁移能力和上调骨痂中CINC-1、CINC-3、LIX、Thymus Chemokine、VEGF,下调TIMP-1表达,进而促进外周血MSCs归巢有关。 |
| 【关键词】骨折愈合 活血化瘀 中草药 归巢 间充质干细胞 |
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| Effect of Taohong Siwu Decoction(桃红四物汤) early intervention on mesenchymal stem cells homing in fracture healing in rats |
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ABSTRACT
Objective: To observe the effects of Taohong Siwu Decoction(桃红四物汤,THSWD) on the mesenchymal stem cells(MSCs) migration,homing number and cytokine expression in callus during the early process of fracture healing,and to explore the mechanism of THSWD on accelerationg fracture healing by regulating the homing of MSCs in rats.
Methods: A rat model of right femoral shaft open fracture was established. Thirty-two 5-week-old male Sprague-Dawley rats,weighting 110 to 130 g,were divided into control group,low-dose group,medium-dose group and high-dose group by using random number table. Distilled water was given to the control group,and the other groups were given Taohong Siwu Decoction. The rats were gavaged twice a day for 5 consecutive days after surgery. Bone volume/tissue volume(BV/TV) and bone mineral density(BMD) were observed using micro-computed tomography (micro-CT) at 21 days after surgery. At 5 days post-fracture,peripheral blood MSCs from THSWD treated and untreated rats were cultured in vitro. Subsequently,the migration ability of MSCs was observed by cell migration assay. The number of MSCs homing to the callus at the early stage of fracture (5 d) was detected by Immunohistochemistry (IHC). Protein chip was used to detect the expression of cytokines in callus.
Results: Micro-CT results showed that BV/TV was higher in the high-dose group than in the medium-dose group (P=0.032),and higher in the medium-dose group than in the low-dose group(P=0.041),with no difference between the control and low-dose group (P=0.651). In addition,there was no difference in BMD between low-dose group and the model group (P=0.671),and lower in the low-dose group than in the medium-dose group(P=0.018),and the medium-dose group was lower than the high-dose group(P=0.008). Cell migration assay showed that THSWD promotes enhanced the migration ability of peripheral blood MSCs. IHC assay revealed that CD45-,CD90+,CD29+ MSCs significantly increased in bone callus after THSWD intervention compared with the control group. Protein chip showed that THSWD promoted the upregulation of CINC-1(×2.91),CINC-3(×1.59),LIX(×1.5),Thymus Chemokine (×2.55),VEGF (×1.22) and the down-regulation of TIMP-1 (×2.98).
Conclusion: THSWD,a representative formula of "promoting blood circulation and removing blood stasis",can significantly accelerate fracture healing,and its mechanism may be related to enhancing the migration ability of peripheral blood MSCs and up-regulating CINC-1,CINC-3,LIX,Thymus Chemokine,VEGF and down-regulating TIMP-1 in bone callus,which promotes the peripheral blood MSCs homing in the early stage of fracture. |
| KEY WORDS Fracture healing Promoting blood circulation and removing blood stasis Drugs,Chinese herbal Homing Mesenchymal stem cells |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 李汪洋,熊辉.桃红四物汤早期干预对大鼠骨折愈合中间充质干细胞归巢的影响[J].中国骨伤,2022,35(4):367~374 |
| 英文格式: | LI Wang-yang,XIONG Hui.Effect of Taohong Siwu Decoction(桃红四物汤) early intervention on mesenchymal stem cells homing in fracture healing in rats[J].zhongguo gu shang / China J Orthop Trauma ,2022,35(4):367~374 |
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