低强度脉冲超声对磷酸三钙磨损颗粒诱导的假体周围骨细胞损伤的影响

发布时间：2018-10-15 11:32

【摘要】：目的:观察低强度脉冲超声(LIPUS)对磷酸三钙(tricalcium phosphate,TCP)磨损颗粒诱导的假体周围骨细胞(osteocyte)损伤的影响,并探讨其可能作用机理。方法:取6~8周龄ICR雄性小鼠30只,随机分为正常组、模型组和LIPUS治疗组,每组10只。模型组和LIPUS治疗组小鼠分别于第1、3、5、7、9和第11周向颅顶注射TCP磨损颗粒建立小鼠颅骨溶解模型。正常组小鼠颅顶皮肤仅接受阴性超声探头按压处理,治疗组小鼠颅顶皮肤接受LIPUS干预治疗3个月,实验结束后取颅骨。采用Micro-CT分析各组小鼠颅骨TCP磨损颗粒植入部位周围溶解情况;应用HE染色比较各组假体周围骨细胞活性;通过流式细胞术定量检测各组假体周围骨细胞凋亡情况;采用免疫印迹技术检测各组假体周围骨细胞牙本质基质蛋白(DMP-1)、骨硬化蛋白(SOST)、葡萄糖调节蛋白78(GRP78)、肌醇依赖酶1α(IRE1α)、X盒结合蛋白(XBP1s)、c-Jun氨基末端激酶(JNK)及磷酸化JNK(p-JNK)等蛋白表达水平。结果:与正常组比较,模型组小鼠假体周围骨细胞活性明显降低,细胞凋亡显著(P0.05),其特征蛋白DMP-1显著下调,SOST明显上调,造成DMP-1/SOST增加(P0.05),且内质网应激通路蛋白GRP78、IRE1α、XBP1s和p-JNK表达水平显著增加(P0.05)。而LIPUS治疗组假体周围骨细胞损伤及内质网应激反应显著减弱,表现为假体周围骨细胞活性、数量和DMP-1/SOST明显增加(P0.05),且IRE1α-XBP1-JNK通路的活化被显著抑制(P0.05)。结论:LIPUS可阻止TCP磨损颗粒诱导的假体周围骨细胞损伤,其作用机制可能与抑制IRE1α-XBP1-JNK信号通路介导的内质网应激反应密切相关。
[Abstract]:Aim: to observe the effect of low intensity pulsed ultrasound (LIPUS) on (osteocyte) damage induced by tricalcium phosphate (tricalcium phosphate,TCP) wear particles and to explore its possible mechanism. Methods: thirty ICR male mice aged 6 weeks and 8 weeks were randomly divided into normal group, model group and LIPUS treatment group with 10 mice in each group. The model group and the LIPUS treatment group were injected with TCP wear granule to the top of the cranium at the 1st and 11th week respectively to establish the model of craniolysis in mice. The skullcap skin in the normal group was only treated with negative ultrasound probe and treated with LIPUS intervention for 3 months. The skull was removed after the experiment. Micro-CT was used to analyze the dissolution around the implanted site of TCP wear particles in the skull of each group; HE staining was used to compare the osteocyte activity around the prosthesis in each group; flow cytometry was used to quantitatively detect the apoptosis of bone cells around the prosthesis in each group. The expressions of dentine matrix protein (DMP-1), osteosclerotic protein (SOST), glucose regulatory protein 78 (GRP78), IRE1 伪), X cassette binding protein (XBP1s), c-Jun amino-terminal kinase (JNK) and phosphorylated JNK (p-JNK) were detected by Western blot. Results: compared with the normal group, the osteocyte activity and apoptosis were significantly decreased in the model group (P0.05). The characteristic protein DMP-1 was significantly down-regulated and the SOST was up-regulated in the model group. DMP-1/SOST was increased (P0.05), and the expression levels of GRP78,IRE1 伪, XBP1s and p-JNK in ER stress pathway were significantly increased (P0.05). In LIPUS treatment group, the injury of periprosthetic bone cells and the stress response of endoplasmic reticulum (ER) were significantly decreased, showing that the activity of osteocytes around the prosthesis, the quantity and DMP-1/SOST were significantly increased (P0.05), and the activation of IRE1 伪-XBP1-JNK pathway was significantly inhibited (P0.05). Conclusion: LIPUS can prevent the injury of periprosthetic bone cells induced by TCP wear particles, and its mechanism may be related to the inhibition of endoplasmic reticulum stress mediated by IRE1 伪-XBP1-JNK signaling pathway.
【作者单位】：绍兴文理学院医学院;
【基金】：浙江省大学生科学创新活动计划(2016R428011) 绍兴市大学生科技创新项目(绍市教高[2015]138号) 国家级大学生创新创业训练计划项目(201610349010) 浙江省自然科学基金(LY17H060007)
【分类号】：R684

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