氨基胍对大鼠体外全骨髓诱导破骨细胞分化的影响

发布时间：2018-03-30 04:20

本文选题：氨基胍　切入点：破骨细胞　出处：《山西医科大学》2017年硕士论文

【摘要】：目的:研究不同浓度氨基胍对大鼠体外全骨髓诱导破骨细胞分化的影响,为骨质疏松性疾病药物治疗提供实验依据。方法:选用全骨髓诱导法培养破骨细胞(Osteoclast,OC),在培养当天各组分别加入浓度为0mM、0.1mM、1mM、2.5mM的氨基胍(Aminoguanidine,AG),于第10天行抗酒石酸酸性磷酸酶(Tartrate-resistant acid phosphatase,TRAP)染色,观察TRAP阳性细胞数的差异;于培养第10天进行扫描电镜检测骨陷窝发生的情况;于培养第10天收集细胞行RT-PCR检测,测定OC特异性标志物核因子-κB受体活化因子(Receptor activator of nuclear,RANK)和组织蛋白酶K(CathepsinK,CK)mRNA的表达量;于培养第10天收集细胞行Western Blot检测OC活化信号通道蛋白P38和P65的表达量,及其测晚期糖基化终末产物(Advanced glycation end products,AGEs)和晚期糖基化终末产物受体(Receptor for advanced glycation end products,AGER)的表达量。结果:TRAP染色观察示:不同浓度AG干预后,能够起到促进OC生成的作用,随AG浓度增加,出现红染细胞数逐渐增多,各组间差异有统计学意义,P0.05;骨吸收实验示:不同浓度AG干预后,能够促进骨陷窝生成;RT-PCR检测结果示:不同浓度AG干预后,能够促进OC形成,随AG浓度的增加,OC特异标志物RANK和CathepsinK的mRNA表达量逐渐增加,各组间差异有统计学意义;Western Blot检测结果示:不同浓度AG干预后,能够促进OC形成,随AG浓度的增加,P38和P65的表达量逐渐增加,同时检测AGEs及其受体AGER的表达量逐渐降低,差异有统计学意义。结论:1.AG可以降低体外全骨髓诱导OC中AGEs和AGER的含量,在本实验中起到了抑制AGEs及其受体AGER的作用,呈剂量依赖性。2.AG能够增加体外全骨髓诱导OC的形成,在本实验浓度范围内促进了OC分化,呈剂量依赖性。
[Abstract]:Objective: to study the effects of different concentrations of aminoguanidine on osteoclast differentiation induced by whole bone marrow of rats in vitro. Methods: osteoclasts of osteoclasts were cultured by whole bone marrow induction method. Aminoguanidine Aminoguanidine (Aminoguanidine) Aminoguanidine (Aminoguanidine) Aminoguanidine (Aminoguanidine) Aminoguanidine (Aminoguanidine) Aminoguanidine (Aminoguanidine) Aminoguanidine (Aminoguanidine) Aminoguanidine (Aminoguanidine) Aminoguanidine (Aminoguanidine) Amin@@. The number of TRAP positive cells was observed, the incidence of bone lacuna was detected by scanning electron microscope on the 10th day of culture, and the RT-PCR was detected by RT-PCR on the 10th day of culture. The expression of nuclear factor 魏 B receptor activator activator of nuclear receptor activator activator of nuclear receptor (RANKK) and cathepsin Knk mRNA were measured, and the expression of OC-activated signal channel proteins P38 and p65 were detected by Western Blot on the 10th day of culture, and the expression of OC-activated signal channel protein p38 and p65 were detected by Western Blot. The expression of advanced glycation end products (ages) and late glycosylation end product receptor (Receptor for advanced glycation end products agers) were measured. The number of red stained cells increased gradually, and the difference was statistically significant (P 0.05). The results of bone absorption test showed that AG at different concentrations could promote the formation of bone lacunae by RT-PCR. With the increase of AG concentration, the mRNA expression of RANK and CathepsinK, the specific markers of OC, increased gradually. The results of Western Blot analysis showed that the different concentrations of AG could promote the formation of OC. With the increase of AG concentration, the expression of P38 and p65 increased, and the expression of AGEs and its receptor AGER decreased gradually. Conclusion: 1. AG can reduce the content of AGEs and AGER in bone marrow induced OC in vitro. In this experiment, AGEs and its receptor AGER were inhibited in a dose-dependent manner. 2. AG could increase the formation of OC induced by whole bone marrow in vitro, and promote the differentiation of OC in a dose-dependent manner.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R580

【参考文献】