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  

Effects of low intensity pulsed ultrasound on the expression of ERS-related Caspase-12 in chondrocytes of rats with knee osteoarthritis

作　　者：刘秋红[1] 孔瑛[1] 李灿阳 张长杰[1] LIU Qiuhong;KONG Ying;LI Canyang(The Second Xiangya Hospital,Central South University,Changsha,Hunan,410011)

机构地区：[1]中南大学湘雅二医院康复科,湖南省长沙市410011

出　　处：《中国康复医学杂志》2020年第10期1172-1180,共9页Chinese Journal of Rehabilitation Medicine

基　　金：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)。

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。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 diffe

关 键 词：低强度脉冲超声 骨关节炎 内质网应激 Caspase-12途径 

分 类 号：R684[医药卫生—骨科学] R49[医药卫生—外科学]