| 刘秋红,孔 瑛,李灿阳,张长杰.低强度脉冲超声对大鼠膝骨关节炎软骨细胞内质网应激相关Caspase-12表达的影响[J].中国康复医学杂志,2020,(10):1172~1180 |
| 低强度脉冲超声对大鼠膝骨关节炎软骨细胞内质网应激相关Caspase-12表达的影响 点此下载全文 |
| 刘秋红 孔 瑛 李灿阳 张长杰 |
| 中南大学湘雅二医院，湖南省长沙市人民中路139号湘雅二医院康复科，410011 |
| 基金项目：湖南省自然科学基金青年项目（2018JJ3729）；湖南省残疾人康复项目（2017XK016） |
| DOI：10.3969/j.issn.1001-1242.2020.10.004 |
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| 摘要： |
| 摘要
目的：通过研究低强度脉冲超声(low intensity pulsed ultrasound， LIPUS)对大鼠膝骨关节炎(osteoarthritis， OA)软骨细胞内质网应激相关Caspase-12表达的影响，探讨低强度脉冲超声对骨关节炎的作用机制。
方法：体外培养大鼠膝关节软骨细胞，并用IL-1β进行骨关节炎造模。取3代以内软骨细胞随机分为空白对照组（A组），OA模型对照组（B组），OA+LIPUS治疗组（C组），OA+磷酸盐缓冲液（PBS）对照组（D组），OA+4-苯基丁酸（4-PBA）组（E组），OA+4-苯基丁酸（4-PBA）+LIPUS治疗组（F组）。超声治疗参数为3MHz，20%，0.1W/cm2，20min/d，治疗7天。采用HE染色观察细胞形态，免疫细胞化学染色检测胞质Col2的表达，Western-blot检测MMP-13、Bax、Caspase-12蛋白水平。
结果：①HE染色结果显示与A组对比，B、D组细胞数目最少，形态最差；C组较B组细胞数目及形态有明显改善；与D组对比，E、F组细胞数目增多，细胞形态可。②Col2免疫细胞化学染色，平均光密度值（AOD 值）进行比较，B组与A组对比AOD 值明显降低；C组与B组对比AOD值有明显的升高；D组、B组比较差异无显著性意义；E组较D组AOD值显著升高；F组、E组AOD值差异无显著性意义。③蛋白印迹检测MMP-13、Bax、Caspase-12蛋白表达水平，B组与A组比较蛋白表达增多（P＜0.05）；D组与B组比较差异无显著性意义；E组与D组比较蛋白表达降低（P＜0.05）；F组与E组比较差异无显著性意义。
结论：低强度脉冲超声可通过抑制骨关节炎软骨细胞内质网应激相关的Caspase-12凋亡途径，减少软骨细胞凋亡，延缓OA的发生发展。 |
| 关键词：低强度脉冲超声 骨关节炎 内质网应激 Caspase-12途径 |
| Effects of low intensity pulsed ultrasound on the expression of ERS-related Caspase-12 in chondrocytes of rats with knee osteoarthritis Download Fulltext |
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| The Second Xiangya Hospital， Central South University， Changsha， Hunan， 410011 |
| Fund Project: |
| Abstract: |
| Abstract
Objective: To investigate the effect of low intensity pulsed ultrasound on osteoarthritis by studying the effects of low intensity pulsed ultrasound on the endoplasmic reticulum stress-related Caspase-12 in rat knee osteoarthritis.
Method: Rat knee articular chondrocytes were cultured in vitro， and osteoarthritis was modeled with IL-1β. The chondrocytes within three passages were randomly divided into blank control group (group A)， OA model control group (group B)， OA+LIPUS treatment group (group C)， OA+phosphate buffer saline control group (group D)， OA+4-phenylbutyric acid group (group E)， OA+4-phenylbutyric acid group+LIPUS treatment group (Group F). Ultrasound treatment parameters were 3HMZ， 20%， 0.1W/cm2， 20min/d， treatment for 7 days. The morphology of the cells was observed by HE staining. The expression of cytoplasmic Col2 was detected by immunocytochemistry. The levels of MMP-13， Bax and Caspase-12 were detected by Western-blot.
Result: ①The results of HE staining showed that compared with group A， the number of cells in group B and group D was the least and the morphology was the worst. The number and morphology of cells in group C were significantly improved compared with group B. Compared with group D， the number of cells in group E and group F increased， and the cell morphology was acceptable. ②Col2 immunocyte staining， average optical density value (AOD value) were compared. The AOD value of group B and group A was significantly lower than that of group A. The AOD value of group C and group B was significantly higher than that of group B. There was no statistical difference between group D and group B. The significance of learning; the AOD value of group E was significantly higher than that of group D; the difference of AOD value between group F and group E was not statistically significant. ③The expression levels of MMP-13， Bax and Caspase-12 protein were detected by Western blotting. The expression of specific protein in group B and group A was increased (P＜0.05). There was no significant difference between group D and group B. The difference between group E and group D was compared. Protein expression was decreased (P＜0.05); there was no significant difference between group F and group E.
Conclusion: Low intensity pulsed ultrasound can reduce the apoptosis of chondrocytes and delay the occurrence and development of OA by inhibiting the stress-associated Caspase-12 apoptosis pathway of osteoarthritis chondrocytes. |
| Keywords:low intensity pulsed ultrasound osteoarthritis endoplasmic reticulum stress Caspase-12 pathway |
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