摘要
目的:用雷公藤甲素(Triptolide,TPL)诱导破骨前体细胞(Osteoclast precursor cells,OCPs)凋亡,评价凋亡抑制蛋白1/2 (cellular inhibitors of apoptosis proteins 1/2,c-IAP1/2)在OCPs凋亡中的参与性。方法:从具有人类类风湿关节炎(Rheumatoid Arthritis,RA)临床症状的肿瘤坏死因子(Tumor necrosis factor,TNF)转基因RA小鼠骨髓中分离、诱导OCPs,分为OCPs组、RANKL/OCPs组、TNF/OCPs组、TPL/RANKL/OCPs组和TPL/TNF/OCPs组。TPL作用于核因子受体激活因子(Receptor activator of nuclear factor,RANKL)或TNF诱导的OCPs,用流式细胞法检测早期细胞凋亡,用Western blot法检测cIAP1/2表达,以及用Luminescence法检测天冬氨酸特异性半胱氨酸蛋白酶3/7 (Aspartate specific cysteine protease 3/7,Caspase3/7)表达。结果:与OCPs组比较,RANKL/OCPs组和TNF/OCPs组中凋亡细胞、Caspase3/7水平显著增加(P <0.05),RANKL/OCPs组中c-IAP1/2蛋白水平显著升高(P <0.05)。与RANKL/OCPs组比较,TPL/RANKL/OCPs组凋亡细胞数、Caspase3/7水平显著升高,c-IAP1/2蛋白水平显著降低(P <0.05)。与TNF/OCPs组比较,TPL/TNF/OCPs组凋亡细胞、Caspase3/7水平显著增加、c-IAP1/2蛋白水平显著降低(P <0.05)。与TPL/RANKL/OCPs组比较,TPL/TNF/OCPs组凋亡细胞显著增加,Caspase3/7水平显著降低,差异有统计学意义(P <0.05)。结论:TPL在OCPs分化过程中可诱导OCPs发生早期凋亡,而c-IAP1/2参与其中,本研究为RA治疗靶点提供实验资料。
Objective:To evaluate the involvement of cellular inhibitors of apoptosis proteins 1/2(c-IAP1/2)in apoptosis of osteoclast precursor cells(OCPs)by using triptolide(TPL)to induce the apoptosis of OCPs.Methods:OCPs were induced after being separated from the bone marrow in tumor necrosis factor(TNF)transgenic mice with clinical symptoms of human rheumatoid arthritis(RA).Divided them into the OCPs group,the RANKL/OCPs group,the TNF/OCPs group,the TPL/RANKL/OCPs group and the TPL/TNF/OCPs group respectively.After TPL acted on receptor activator of nuclear factor(RANKL)or TNF to induce OCPs,detected early apoptosis via flow cytometry,detected the expression of cellular inhibitors of apoptosis protein(cIAP1/2)via Western blot,and detected the expression of aspartate specific cysteine protease 3/7(caspase3/7)via luminescence method.Results:Comparing with the OCPs group,the apoptotic cells and levels of caspase3/7 in the RANKL/OCPs group and the TNF/OCPs group were significantly increased(P<0.05),and levels of c-IAP1/2 in the RANKL/OCPs group were significantly higher(P<0.05).Comparing with the RANKL/OCPs group,the apoptotic cellsand levels of caspase3/7 in the TPL/RANKL/OCPs group were significantly increased,and levels of c-IAP1/2 were significantly decreased(P<0.05).Comparing with the TNF/OCPs group,the apoptotic cells and levels of caspase3/7 in the TPL/TNF/OCPs group were significantly increased,and levels of c-IAP1/2 were significantly decreased(P<0.05).Comparing with the TPL/RANKL/OCPs group,the apoptotic cells in the TPL/TNF/OCPs group were significantly increased,and the levels of caspase3/7 were significantly decreased,difference being significant(P<0.05).Conclusion:TPL can induce the early apoptosis of OCPs during differentiation,with the involvement of c-IAP1/2.This study provides experimental data for therapeutic target of RA.
作者
蔡灯塔
王新春
王景春
李宝印
CAI Dengta;WANG Xinchun;WANG Jingchun;LI Baoyin
出处
《新中医》
CAS
2019年第1期1-5,共5页
New Chinese Medicine
基金
国家自然科学基金项目(81471558)
