摘要
背景:成骨细胞数量减少或凋亡进程加速,使骨形成少于骨吸收可导致骨质疏松的发生。肿瘤坏死因子α在去势大鼠或绝经后骨质疏松患者骨组织中呈高表达,在体外可刺激成骨细胞凋亡。健骨颗粒可降低骨质疏松模型鼠血清肿瘤坏死因子α水平。目的:验证健骨颗粒含药血清对肿瘤坏死因子α诱导的鼠成骨细胞凋亡的影响。设计、时间及地点:随机对照动物实验,于2004-07/2006-12在福建中医学院骨伤系分子生物学实验室完成。材料:3月龄SD雌性大鼠40只用于制备含药血清,24hSD乳鼠5只用于培养成骨细胞。健骨颗粒由煅狗骨、淫羊藿、山茱萸、党参等药味组成,原药材由福建省药材公司提供,福建中医药研究院中试车间加工制备,每克颗粒含原生药2.9g。方法:用酶消化法培养成骨细胞。随机将40只SD大鼠分为高、中、低剂量含药血清组和生理盐水对照组,每组10只,分别将健骨颗粒按4,2,1g/(kg·d)的药量灌服,对照组每天灌服2mL生理盐水。连续给药7d后腹主动脉取血得到健骨颗粒含药血清,将含药血清作用于成骨细胞48h后,再用肿瘤坏死因子α诱导24h。运用TUNEL技术、流式细胞术、免疫细胞化学结合图像分析方法,检测成骨细胞凋亡情况。主要观察指标:①成骨细胞凋亡指数。②成骨细胞凋亡峰。③成骨细胞Bcl-2、Bax表达。结果:用肿瘤坏死因子α诱导的各组成骨细胞均出现凋亡现象,其中中、高剂量含药血清组的细胞凋亡指数和凋亡率明显低于其他组,而Bcl-2/Bax灰度比值却较其他组高(P<0.05),Bcl-2/Bax灰度比值与凋亡指数呈负相关(r=-0.757,P<0.01)。结论:健骨颗粒对肿瘤坏死因子α诱导的成骨细胞凋亡有一定的保护作用,可能通过提高Bcl-2的表达来实现。
BACKGROUND： Reducing the number of osteoblast or accelerating the apoptosis will lead to bone formation less than resorption, ultimately result in ostecporosis. Tumor necrosis factor-α （TNF-α） can stimulate the osteoblast apoptosis in vitro, which was highly expressed in castrated rats or postmenopause women with osteoporosis. However, the Jiangu granula can decrease the TNF-o level of rats with osteoporosis. OBJECTIVE： To investigate the effect of Jiangu granula on the apoptosis of rat osteoblast induced by TNF-α. DESIGN, TIME AND SETTING： A randomized controlled experiment of animal was performed at the Laboratory of Molecular Biology, Department of Orthopedics ＆ Grammatology, Fujian College of Traditional Chinese Medicine from July 2004 to December 2006. MATERIALS： A total of 40 SD rats with 3-month-old were prepared for medicated serum. Five neonatal rats （24 hours） were cultured osteoblasts. Jiangu granula （contains： calcined dog bone, epimedium, Fructus Corni and codonopsis pilosula） were provided by the Fujian Provincial Medicinal Materials Company followed by prepared by Fujian Academy of Traditional Chinese Medicine. There was 2.9 g crude drug in per gramme of granula. METHODS： Primary cells were isolated by enzymatic digestion method. Forty rats were randomly divided into the medicated serum with high, medium and low dose, and control groups, with 10 animals in each group. Rats in the medicated serum groups were drenched Jiangu granula with 4, 2, 1 g/（kg.d）, respectively. Rats in the control group were drenched 2 mL normal saline per day. After 7 days, blood from abdominal aorta was obtained, prepared for medicated serum and cultured with osteoblasts for 46 hours, followed by 24 hours TNF-α induction. TUNEL technology, flow cytometry and immunochemical staining combined with imaging analysis process were used to analysis apoptotic status. MAIN OUTCOME MEASURES： The apoptotic index, apoptotic peak, and the Bcl-2, Bax expression of osteoblasts were observed. RESULTS：
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第7期1227-1231,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
国家自然科学基金项目(30572402)
陈可冀中西医结合发展基金项目(CKJ2008004)
福建省中西医结合老年性疾病重点实验室开放课题(2008J1004-26)~~
