摘要
背景:抗骨增生胶囊在治疗骨关节疾病方面有很好的疗效,基础研究多集中于颈椎病等骨关节疾病上。目的:拟观察抗骨增生胶囊含药血清对白细胞介素1β引起的软骨细胞增殖凋亡及基质金属蛋白酶分泌的影响。设计、时间及地点:分组对照体外细胞学实验,于2007-03/09在河北医科大学解剖教研室细胞培养中心完成。材料:新生1周SD大鼠,用于培养软骨细胞;体质量200g左右SD大鼠10只,以中药水溶液灌胃后定时取血制备含药血清。方法:采用胶原酶NB4消化法从大鼠关节软骨中分离出软骨细胞并进行原代和传代培养,实验分为3组:空白血清组:90%L-DMEM+10%空白血清,每日给予同等量的生理盐水;白细胞介素1β组(90%DMEM+10%空白血清+白细胞介素1β);中药血清+白细胞介素1β组(90%DMEM+10%中药血清+白细胞介素1β)。白细胞介素1β加入量为5μg/L,培养72h。将实验用细胞消化传代后制成单细胞悬液,以1×108L-1的密度接种于96孔培养板,100μL/孔,每组6孔。主要观察指标:倒置显微镜下观察中药含药血清对软骨细胞增殖的影响;四甲基偶氮唑盐比色法检测其各孔吸光度变化;流式细胞仪分析凋亡细胞百分率;基质金属蛋白酶13Elisa试剂盒检测不同组基质金属蛋白酶13分泌情况。结果:在正常培养基条件下,细胞形态均匀,死亡脱落细胞较少。空白血清组软骨细胞的增殖速度缓慢,细胞较稀疏。白细胞介素1β组细胞明显稀疏,部分细胞胞浆中空泡增多,细胞核形态不规则。在中药血清培养基作用下,软骨细胞增殖的速度较白细胞介素1β组快,胞体显著增大,细胞核形态规则。白细胞介素1β组细胞增殖数量减少,凋亡细胞百分率增加,其培养的细胞上清液中基质金属蛋白酶13含量升高,抗骨增生胶囊含药血清可对抗白细胞介素1β对软骨细胞的影响。结论:体外条件下,抗骨增生胶囊含药血清能够抑制白细胞介�
BACKGROUND： Kanggu Zengsheng capsules exhibit efficacy in treating joint disease. There are many basic studies focusing on cervical syndrome or other joint diseases. OBJECTIVE： To observe the seropharmacological effects of Kanggu Zengsheng capsules on the proliferation, apoptosis and secretion of matrix metalloproteinase 13 （MMP-13） of chondrocytes induced byinterleukin-1β （IL-1β ）. DESIGN, TIME AND SETTING： The in vitro cytology trial was performed at the Cell Laboratory of Department of Anatomy of Hebei Medical University from March to September 2007. MATERIALS： One 1-week-old SD rat was prepared for chondrocyte culture; the serum of ten SD rats weighing 200 g was obtained after intragastric administration with medicine water solution. METHODS： Articular chondrocytes were obtained from the cartilage of 1-week SD rat using digestion method by collagenase NB4, followed by primary culture and subculture of chondrocytes. It was divided into 3 groups： control group, 90%L-DMEM＋ 10% serum, and normal saline; IL-1β group, 90% DMEM＋ 10% serum＋5 μ g/L IL-1 β ; and Kanggu Zengsheng capsule-conditioned serum ＋IL-1 β group, 90%DMEM＋ 10% medicine-conditioned serum＋5 μ g/L IL-1β ）. After 72 hours incubation, the chondrocytes were digested into single cell suspension. Then they were incubated in the 96-well culture plate with the density of 1×10^8 L^-1 , 100 μL per well and six wells for per group. MAIN OUTCOME MEASURES： The seropharmacological effects of Kanggu Zengsheng capsules on the proliferation of chondrocytes were observed by inverted microscope. The absorbance was measured by methyl thiazolyl tetrazolinm method （MTT）. The apoptosis ratio was analyzed by flow cytometry. The concentration of MMP- 13 on the superoatants was measured by Elisa kit. RESULTS： In normal condition, the chondrocytes aligned closely with uniform shape, and few died cells were seen. In control group, the proliferation was slow, and the chondrocytes were dispersed. In IL-1 β group, the cells ali
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第24期4642-4646,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
河北省科学技术研究与发展指导计划项目(052761617)~~
