低强度脉冲超声通过初级纤毛调控软骨细胞  

Low-intensity pulsed ultrasound regulates chondrocytes through primary cilia

作　　者：武沙 周海琪 凌慧贤 雒紫寓 孙雨嫣 Ngo Thai Nam Anh 张长杰[1] 孔瑛[1] WU Sha;ZHOU Haiqi;LING Huixian;LUO Ziyu;SUN Yuyan;Ngo Thai Nam Anh;ZHANG Changjie;KONG Ying(Department of Rehabiltation,the Second Xiangya Hospital,Central South University,Changsha,Hunan 40000,P.R.China)

机构地区：[1]中南大学湘雅二医院康复科,长沙410000

出　　处：《华西医学》2023年第6期835-842,共8页West China Medical Journal

基　　金：国家自然科学基金(82060418);湖南省自然科学基金(2021JJ30933)。

摘　　要：目的 探讨低强度脉冲超声(low-intensity pulsed ultrasound, LIPUS)对白细胞介素(interleukin,IL)-1β干预后的C57小鼠软骨细胞的合成代谢、细胞凋亡、纤毛内转运蛋白88(intraflagellar transport 88, IFT88)表达的影响,以及LIPUS修复软骨与初级纤毛的相关性。方法 分别对小鼠软骨细胞进行IL-1β干预、LIPUS干预、携带IFT88短发卡RNA的慢病毒转染,分组包括正常软骨细胞组(N组)、IL-1β干预软骨细胞组(OA组)、IL-1β干预软骨细胞+LIPUS组(OA+U组)、慢病毒沉默IFT88+IL-1β干预软骨细胞组(KO+OA组)、慢病毒沉默IFT88+LIPUS+IL-1β处理软骨细胞组(KO+OA+U组)。利用实时聚合酶链反应和免疫荧光实验检测各组软骨合成基质Ⅱ型胶原、蛋白聚糖、初级纤毛的表达,利用流式细胞术检测细胞凋亡。所有实验数据采用GraphPad Prism 9.5软件进行统计分析。结果 OA+U组较OA组的小鼠软骨细胞Ⅱ型胶原、蛋白聚糖表达上升,凋亡降低,初级纤毛发生率上升,差异有统计学意义(P<0.05);而KO+OA+U组较OA+U组的小鼠软骨细胞Ⅱ型胶原、蛋白聚糖表达下降,凋亡上升,差异有统计学意义(P<0.05)。结论 LIPUS可减少IL-1β干预后C57小鼠软骨细胞的凋亡,增加软骨细胞基质中Ⅱ型胶原、蛋白聚糖的表达,且该作用与初级纤毛有关。Objective To explore the effects of low-intensity pulsed ultrasound(LIPUS)on anabolism,apoptosis and intraflagellar transport 88(IFT88)expression in mouse chondrocytes after interleukin(IL)-1βintervention,and the correlation of cartilage repairment by LIPUS with primary cilia.MethodsIL-1βintervention,LIPUS intervention and lentiviral carrying IFT88-specifific short hairpin RNA(sh-IFT88)transfection were performed on mouse chondrocytes,respectively.The groups included:normal chondrocyte group(N group),chondrocyte after IL-1βintervention group(OA group),chondrocyte after IL-1βintervention+LIPUS group(OA+U group),sh-IFT88+IL-1βintervention chondrocyte group(KO+OA group),and sh-IFT88+LIPUS+IL-1βtreated chondrocyte group(KO+OA+U group).Real-time polymerase chain reaction and immunofluorescence were used to determine the expression of collagen Ⅱ,aggrecan,and primary cilia,and apoptosis was measured by flow cytometry.All experimental data were statistically analyzed using the GraphPad Prism 9.5 software.Results The expression of collagen Ⅱ and aggrecan increased,the apoptosis decreased,and the incidence of primary cilia in chondrocytes of mice increased in the OA+U group compared with those in the OA group(P<0.05).The collagen I and aggrecan expression decreased and the apoptosis increased in the KO+OA+U group compared with those in the OA+U group(P<0.05).Conclusion LIPUS can reduce the apoptosis of chondrocytes in C57 mice after IL-1βintervention,and increase the expression of collagen Ⅱ and aggrecan in chondrocyte matrix,and the effect is related to primary cilia.

关 键 词：低强度脉冲超声 初级纤毛 白细胞介素-1Β 软骨细胞 

分 类 号：R684.3[医药卫生—骨科学] R445.1[医药卫生—外科学]