| 摘要: |
| 目的 探究冬凌草甲素对海水冲溺伤骨折愈合的影响及作用机制。方法 取72只6～8周龄雄性C57BL/6小鼠构建小鼠海水冲溺伤骨折模型，并将其随机分为骨折组、骨折＋冬凌草甲素组、冲溺伤骨折组和冲溺伤骨折＋冬凌草甲素组，每组18只。术后14 d拍摄股骨X线片并在术后21 d通过股骨机械强度试验观察冬凌草甲素对骨折愈合影响的表型变化。术后14 d取小鼠骨痂组织，采用qPCR和蛋白质印迹法检测M1型巨噬细胞标志分子诱导型一氧化氮合酶（iNOS），M2型巨噬细胞标志分子CD206，炎症因子环氧合酶2（COX2）、IL-1β，促凋亡因子cleaved caspase 3、Bcl-2相关X蛋白（Bax）及抗凋亡因子Bcl-2的表达水平。术后7 d，采用抗酒石酸酸性磷酸酶（TRAP）染色观察破骨细胞活化情况。结果 X线片及股骨机械强度试验结果显示冲溺伤骨折组小鼠骨折愈合减弱，而冬凌草甲素改善了冲溺伤骨折愈合并提高了股骨承受的扭转力（P均＜0.01）。qPCR和蛋白质印迹法检测结果显示，与冲溺伤骨折组相比，冲溺伤骨折＋冬凌草甲素组小鼠骨痂组织中iNOS、COX2、IL-1β、cleaved caspase 3、Bax的mRNA和蛋白表达水平均降低，CD206、Bcl-2的mRNA和蛋白表达水平均升高（P＜0.05，P＜0.01）。TRAP染色结果显示，与冲溺伤骨折组相比，冲溺伤骨折＋冬凌草甲素组小鼠软骨-骨边界附近破骨细胞数量减少（P＜0.01）。结论 冬凌草甲素通过增强抗炎性的M2型巨噬细胞极化和抑制促炎性的M1型巨噬细胞极化减轻骨折后的炎症反应、减少细胞凋亡、抑制破骨细胞过度激活，从而促进海水冲溺伤骨折愈合。 |
| 关键词: 海水浸泡 开放性骨折 冬凌草甲素 骨折愈合 极化 炎症 破骨细胞 细胞凋亡 |
| DOI：10.16781/j.CN31-2187/R.20211116 |
| 投稿时间：2021-11-03修订日期：2022-07-01 |
| 基金项目: |
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| Effect and mechanism of oridonin on healing of bone fracture with seawater immersion |
| LI Fu-dong,KONG Fan-qi,SUN Kai-qiang,SUN Jing-chuan,SHI Jian-gang* |
(The Second Ward, Department of Spinal Surgery, The Second Affiliated Hospital of Naval Medical University (Second Military Medical University), Shanghai 200003, China
*Corresponding author) |
| Abstract: |
| Objective To investigate the effect and mechanism of oridonin (Ori) on the healing of bone fracture with seawater immersion (BFSI).Methods A total of 72 male C57BL/6 mice (6-8 weeks old) were used to construct BFSI model mice,and then they were randomly divided into bone fracture (BF) group,BF+Ori group,BFSI group,and BFSI+Ori group,with 18 mice in each group.X-ray of femur was taken 14 d after operation and femur mechanical strength test was performed to observe the phenotypic changes of Ori on fracture healing 21 d after operation.The callus tissues of mice were harvested 14 d after operation.The expression levels of the M1 macrophage marker (inducible nitric oxide synthase[iNOS]),M2 macrophage marker CD206,inflammatory factors (cyclooxygenase 2[COX2]and interleukin[IL]-1β),pro-apoptotic factors (cleaved cysteine aspartic acid specific protease[caspase 3]and Bcl-2-associated X protein[Bax]) and anti-apoptotic factor (B-cell lymphoma 2[Bcl-2]) were detected by quantitative polymerase chain reaction (qPCR) and Western blotting.The activation of osteoclasts was observed by tartrate-resistant acid phosphatase (TRAP) staining 7 d after operation.Results The results of X-ray and femur mechanical strength test showed that the fracture healing was significantly weakened in the BFSI group,while Ori significantly improved the healing of BFSI and increased the torsion force of the femur (all P<0.01).The results of qPCR and Western blotting showed that the mRNA and protein expression of iNOS,COX2,IL-1β,cleaved caspase 3 and Bax in the callus tissues of mice was significantly decreased,while the mRNA and protein expression of CD206 and Bcl-2 was significantly increased in the BFSI+Ori group compared with the BFSI group (P<0.05,P<0.01).The results of TRAP staining showed that the number of osteoclasts in the vicinity of the cartilage bone boundary was significantly decreased in the BFSI+Ori group compared with the BFSI group (P<0.01).Conclusion Oridonin can alleviate the inflammatory responses after fracture,reduce cell apoptosis,and inhibit the overactivation of osteoclasts,so as to promote the healing of BFSI by enhancing the polarization of anti-inflammatory M2 macrophages and inhibiting the polarization of pro-inflammatory M1 macrophages. |
| Key words: seawater immersion open fractures oridonin fracture healing polarization inflammation osteoclasts apoptosis |
