GC induced osteoporosis
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克老素抑制地塞米松诱导的MC3T3-E1成骨细胞凋亡黄倩;李宝善;梁霄;刘丹;马厚勋【摘要】目的探究体外转染克老素(Klotho,KL)基因对地塞米松(dexamethason,DEX)诱导的MC3T3-E1成骨细胞凋亡的影响.方法用携带KL基因的重组腺病毒(Ad-KL)及携带绿色荧光蛋白基因的重组腺病毒(Ad-GFP)感染细胞,并建立DEX诱导凋亡的细胞模型.运用荧光倒置显微镜观察重组腺病毒的转染效果,qPCR 与 Western blot 法分析 KL mRNA和蛋白表达情况;CCK-8法检测不同浓度DEX作用于MC3T3-E1细胞后的存活率及各研究组细胞的生存情况;流式细胞仪分析各研究组细胞的凋亡率;qPCR 与 Western blot分析凋亡标志物的mRNA、蛋白表达;免疫荧光分析caspase-9的荧光蛋白表达情况.结果重组腺病毒成功感染MC3T3-E1成骨细胞,KL组和KL+DEX组的KL mRNA与蛋白表达量较非转染组明显升高.CCK-8法检测出DEX的适宜干预浓度为2.0 mmol·L-1.DEX 组、GFP+DEX组较其余组细胞存活率明显降低、凋亡率明显增加,Bax、Bcl-2的mRNA与蛋白表达分别表现出增加和降低趋势;KL组细胞存活率、凋亡率,以及Bcl-2、Bax mRNA 与蛋白表达较DEX组及KL+DEX组均明显改善.结论大剂量DEX的运用能够诱发MC3T3-E1成骨细胞凋亡,上调KL表达具有抵抗DEX诱导成骨细胞凋亡的作用,提示上调KL表达可改善糖皮质激素诱导性骨质疏松,为大剂量使用糖皮质激素所致医源性骨质疏松的治疗提供新的靶点.%Aim To explore the effect of Klotho (KL) gene transfection on the apoptosis of MC3T3-E1 osteo-blasts induced by dexamethasone(DEX). Methods MC3T3-E1 osteoblasts were transfected by recombinant adenovirus containing KL gene(Ad-KL) and recombi-nant adenovirus containing green fluorescent protein (GFP) gene(Ad-GFP). The apoptosis model was con-structed. Thetransfection efficiency of Ad-KL and Ad-GFP in cells were observed using inverted fluorescent microscope, and the level of KL mRNA and protein was detected by qPCR and Western blot,respectively. The cell viability after different concentrations of DEX acting on the cells and the viability of every research group were determined by cell counting kit-8 (CCK-8) assay. The apoptotic rate was evaluated by flow cytom-etry. The level of mRNA and protein was analyzed by qPCR and Western blot, respectively. The level of caspase-9 protein was detected by immunofluorens-cence assay. Results Cells were transfected by Ad-KL and Ad-GFP successfully. KL group and KL +DEX group had higher level of KL mRNA and protein than that in other groups. The optimum concentration of DEX was 2.0 mmol·L-1. When DEX acting on the cells, the cells viability decreased and apoptotic rate increased obviously in DEX group and GFP + DEX group. The level of Bax mRNA and protein presented a upward trend in DEX group and GFP +DEX group, while the level of Bcl-2 mRNA and protein was oppo-site. But after KL transfecting MC3T3-E1 osteoblasts, the markers described above in KL group had more dramatic improvement than in DEX group and KL +DEX group. Conclusions High-dosage DEX can in-duce the apoptosis ofMC3T3-E1 osteoblasts, and the pro-apoptosis effect of high-dosage DEXin MC3T3-E1 osteoblasts can be suppressed by up-regulating KL gene expression level, suggesting that the glucocorticoid-in-duced osteoporosis might be improved by up-regulating KL gene expression level, and it may be a new target for the treatment of latrogenic osteoporosis induced by high-dosage glucocorticoid in clinic.【期刊名称】《中国药理学通报》【年(卷),期】2018(034)004【总页数】7页(P570-576)【关键词】克老素;地塞米松;MC3T3-E1成骨细胞;凋亡;Bcl-2;Bax;caspase-9【作者】黄倩;李宝善;梁霄;刘丹;马厚勋【作者单位】重庆医科大学附属第一医院老年病科,重庆 400016;重庆市急救医疗中心老年病科,重庆 400014;重庆医科大学附属第一医院老年病科,重庆 400016;重庆医科大学附属第一医院老年病科,重庆 400016;重庆医科大学附属第一医院老年病科,重庆 400016【正文语种】中文【中图分类】R329.25;R373.9;R394.2;R681;R977.11;R977.6地塞米松(dexamethasone, DEX)等糖皮质激素作为临床上极为常用的抗炎药、免疫抑制药,广泛用于诸如类风湿性关节炎、炎症性肠病、慢性阻塞性肺疾病等疾病的治疗。
骨质疏松症的药物治疗冯颖瑜【摘要】骨质疏松症是一种骨退行性病变,其发生主要与骨吸收和骨重建失衡有关.根据骨质疏松症的发病机制的不同,临床治疗药物主要分为基本补充剂、抗骨吸收药物和促骨形成药物.钙剂和维生素D是药物治疗的基础;抗骨吸收药物包括双磷酸盐、雌激素及其受体调节剂、降钙素;甲状旁腺激素为促骨形成药物;锶盐、维生素K2等兼有抗骨吸收和促骨形成的作用.特殊人群以及不同类型的骨质疏松症治疗要注意药物的适用性和针对性.【期刊名称】《医学综述》【年(卷),期】2014(020)001【总页数】5页(P105-109)【关键词】骨质疏松症;治疗;药物;抗骨吸收剂;促骨形成剂【作者】冯颖瑜【作者单位】中山大学附属第一医院,内分泌科,广州,510080【正文语种】中文【中图分类】R589.9;R681骨质疏松症是一种骨退行性病变,以老年人和绝经后妇女多见。
随着人类寿命的延长和社会老龄化的发展,骨质疏松所导致的骨折在大大增加老年人病残率和病死率的同时,明显加重了社会公共卫生的经济负担。
临床上,骨质疏松症作为一种常见合并症,已成为各专科医师时常需要面对的、不可忽视的问题。
本文就骨质疏松症的药物治疗包括常见用药、联合用药及特殊人群用药等方面进行综述。
1 骨质疏松症的基本治疗措施针对骨质疏松症和骨折危险因素的基本治疗措施包括:① 调节生活方式(如进食富含钙、蛋白质的饮食,适当户外运动和日照,戒烟、酒,慎用影响骨代谢的药物,加强自身和环境保护,采取各种预防跌倒的措施);② 补充骨健康基本需要剂(适量的钙剂和维生素D)。
一旦基本措施不能很好的改善骨质疏松症症状或患者存在高骨折风险,需要考虑进行抗骨质疏松药物治疗。
2 抗骨质疏松临床药物分类2.1 基本补充剂2.1.1 钙剂钙剂是保证骨健康的基本补充剂,适量的钙可减缓骨丢失,改善骨矿化。
钙剂补充治疗已被证明对绝经后和老年性骨质疏松症有益。
单独钙剂补充治疗可提高绝经后妇女的骨密度,轻微降低骨折发生风险,与维生素D联合应用效果更明显[1]。