摘要
目的探讨大鼠骨髓间充质干细胞(BMSCs)来源的外泌体对髁突软骨细胞蛋白多糖含量表达的影响,为骨关节病的防治提供基础。方法研究时间为2019年1月至2021年1月(其中动物实验部分2014年做)。实验组使用含有骨髓间充质干细胞来源的外泌体进行细胞培养,对照组使用低糖培养基进行细胞培养;收取培养第3、7、14天的细胞,酶标仪下检测髁突软骨细胞氨基聚糖(GAG)含量的表达,荧光酶标仪检测髁突软骨细胞DNA含量,比较单位DNA中氨基聚糖含量的变化(GAG/DNA);反转录聚合酶联反应分析第7、14天两组细胞GAG基因表达情况。两组比较采用独立样本t检验,进行统计学分析。结果酶标仪检测结果示髁突软骨GAG含量在第3天实验组高于对照组[(0.95±0.03)μg/ml比(0.58±0.02)μg/ml],差异有统计学意义(t=10.36,P<0.001);第7天实验组高于对照组[(2.68±0.06)μg/ml比(1.63±0.04)μg/ml],差异有统计学意义(t=12.01,P<0.001);第14天实验组高于对照组[(3.59±0.07)μg/ml比(1.86±0.07)μg/ml],差异有统计学意义(t=10.61,P<0.001)。荧光酶标仪显示第3天实验组DNA含量显著高于对照组[(30.14±9.59)μg/ml比(23.45±3.33)μg/ml],差异有统计学意义(t=3.625,P=0.037)。GAG/DNA比值在第3天实验组高于对照组[(0.0320±0.0071)比(0.0230±0.0035)](t=4.921,P=0.018);第7天实验组高于对照组[(0.0630±0.0065)比(0.0390±0.0067)](t=17.78,P<0.001);第14天实验组高于对照组[(0.0630±0.0073)比(0.0320±0.0058)](t=15.59,P<0.001)。RT-PCR结果显示GAG mRNA在第7天实验组高于对照组[(2.610±0.018)比(1.000±0.150)](t=12.31,P<0.001),第14天实验组高于对照组[(5.560±0.018)比(2.550±0.090)](t=14.78,P<0.001)。结论BMSCs来源的外泌体促进髁突软骨细胞蛋白多糖含量的表达。
Objective To provide evidences for the prevention and treatment of osteoarthrosis by studying the effect of exosomes derived from rat bone marrow mesenchymal stem cells on the expression of glycosaminoglycan in condyle chondrocytes.Methods The study was from January 2019 to January 2021(the rat experiment was did in 2014).In the experimental group,the exosomes(Exos)extracted from rat bone mesenchymal stem cells(BMSCs)were used to culture rat condyle chondrocytes.In the control group,the L-DMEM was used to culture the cells.The cells were collected on day 3,7,and 14 of the culture.The expression of glycosaminoglycan(GAG)was tested by the enzyme label analyzer,and the DNA contents of the condylar chondrocytes by the fluorescein microplate.The GAG/DNA were analyzed and compared between the two groups.The gene expressions of GAG on day 7 and 14 of the two groups were analyzed by reverse transcription-polymerase chain reaction(RT-PCR).Comparisons between the two groups were performed by independent-sample t test for statistical analysis.Results The results showed that the expressions of GAG in the experimental group were higher than those in the control group on day 3,7,and 14[(0.95±0.03)μg/ml vs.(0.58±0.02)μg/ml,(2.68±0.06)μg/ml vs.(1.63±0.04)μg/ml,and(3.59±0.07)μg/ml vs.(1.86±0.07)μg/ml],with statistical differences(t=10.36,P<0.001;t=12.01,P<0.001;and t=12.01,P<0.001).The DNA content of the condylar chondrocytes in the experimental group was higher than that in the control group on day 3[(30.14±9.59)μg/ml vs.(23.45±3.33)μg/ml],with a statistical difference(t=3.625,P=0.037).The GAG/DNA in the experimental group were higher than those in the control group on day 3,7,and 14[(0.0320±0.0071)vs.(0.0230±0.0035),t=4.921,P=0.018;(0.0630±0.0065)vs.(0.0390±0.0067),t=17.78,P<0.001;(0.0630±0.0073)vs.(0.0320±0.0058),t=15.59,P<0.001].The results of RT-PCR indicated that the mRNA expressions of GAG in the experimental group were higher than those in the control group on day 7 and 14[(2.610±0.018)vs.(1.000±0.150),t=12.31,P<0.001;(5.560±0.018)vs.(2.550±0.090),t=14.78,P<0.001].Conclusion Exosomes derived from rat bone marrow mesenchymal stem cells can promote the expression of proteoglycan in condyle chondrocytes.
作者
邢超
徐灵巧
叶钟泰
张志光
Xing Chao;Xu Lingqiao;Ye Zhongtai;Zhang Zhiguang(Department of Stomatology,People's Hospital of Bao'an District,Shenzhen 518101,China;Department of Oral and Maxillofacial Surgery,Hospital of Stomatology,Guanghua School of Stomatology,Sun Yat-sen University,Guangzhou 510055,China)
出处
《国际医药卫生导报》
2021年第23期3678-3683,共6页
International Medicine and Health Guidance News
基金
深圳市宝安区医疗卫生基础研究项目(2020JD351)。
