摘要
目的 观察不同浓度人参皂苷Rg1在体外培养条件下对正常人乳腺脂肪来源干细胞（HBASCs）增殖和成骨分化的影响.方法 体外分离培养HBASCs传至第3代后,按2×104个/孔接种至96孔板,分别加入含10-8、10-7、10-6、10-5和10-4 mol/L Rg1的成骨诱导培养基中进行培养,分别记为B、C、D、E和F组;另设立对照组,仅加入等量标准成骨诱导培养基,记为A组.采用细胞计数试剂盒（CCK-8）检测Rg1促进HBASCs的增殖作用并绘制细胞生长增殖曲线.通过碱性磷酸酶（ALP）试剂盒测定细胞ALP活性,放射免疫法检测骨钙素（OCN）和骨桥蛋白（OPN）的含量,茜素红染色观察矿化钙化结节形成能力.结果 10-8 mol/L Rg1可有效促进HBASCs增殖;随着Rg1浓度的增加,促细胞增殖活性逐渐增强,Rg1浓度为10-6 mol/L时细胞增殖活性最强,当Rg1浓度增加至10-4 mol/L,则表现为明显的抑制细胞增殖作用.Rg1呈剂量依赖性促进HBASCs中ALP活性、OCN和OPN的表达.培养7d后,A组的ALP活性的表达分别为0.13 ±0.02,其余5组（B、C、D、E和F组）依次为0.32±0.03、0.58±0.06、0.82±0.09、1.04±0.10、1.12±0.11,各实验组与对照组比较,P均＜0.05,培养14d后活性进一步提高,A组为0.27±0.03,其余5组依次为0.51±0.05、0.69±0.07、0.95±0.11、1.13±0.13、1.18±0.14,各实验组与对照组比较,P均＜0.05.A组在第14天时的OCN及OPN表达量分别为（0.16±0.02） ng/ml和（0.21 ±0.03） ng/ml,其余5组依次为（0.25±0.03） ng/ml及（0.35±0.04） ng/ml、（0.55±0.06） ng/ml及（0.59±0.06） ng/ml、（0.71±0.08） ng/ml及（0.84±0.09） ng/ml、（0.94±0.10） ng/ml及（1.16±0.12） ng/ml、（0.98±0.11） ng/ml及（1.19±0.12） ng/ml;在第21天时A组OCN及OPN表达量分别为（0.21 ±0.03） ng/ml和（0.33±0.04） ng/ml,其余5组依次为（0.44±0.04） ng/ml及（0.57±0.06） ng/ml、（0.73±0.08） ng/ml及（0.76±0.08） ng/ml、（0.92±0.10） ng/ml及（0.97±0.11） ng/ml、（1.05
Objective To explore the effects of different concentrations of ginsenoside Rg1 on the proliferation and osteogenic differentiation of human breast adipose-derived stem cells （HBASCs） in vitro.Methods HBASCs were isolated and pass to third generation,then seeded HBASCs （2 × 104 cells/well） to 96-well plates cultured with 10-8,10-7,10-6,10-5 and 10-4 mol/L ginsenoside Rg1 in osteogenic induction medium.recorded as B,C,D,E and F group,A group for control group.Using CCK-8 method to detect the proliferation and osteogeuic differentiation of HBASCs a promoted by ginsenoside Rg1 and draw the growth curve of cell proliferation.Alkaline phosphatase activity of the cells was measured by alkaline phosphatase kit,the concentrations of osteocalcin and osteopont were detected by radioimmunoassay,and mineralization calcified nodules were observed by alizarin red staining.Results 10-8 mol/L ginsenoside Rg1 can promote the proliferation HBASCs,the ability of cell proliferation increase as the rising concentrations of ginsenosides Rg1 and reach its peak when co-culture with 10-8 mol/L ginsenoside Rg1,while inhibited in concentration of ginsenosides Rg1 to 10-4 mol/L.Ginsenoside Rg1 dose promote osteogenesis differentiation of HBASCs by raising the,expression of osteocalcin （OCN） and osteopontin （OPN）.7 d after co-culture,theactivity of alkaline phosphatasein group A was 0.13 ± 0.02,While the group B-F were 0.32 ±0.03,0.58 ±0.06,0.82 ±0.09,1.04 ±0.10,1.12 ±0.11,which significant greater than group A,P 〈 0.05.At 14d,the activity of alkaline phosphatase in group A was 0.27 ± 0.03,While the group B-F were 0.51 ±0.05,0.69 ±0.07,0.95 ±0.11,1.13 ±0.13,1.18 ±0.14,which significant greater than group A,P〈 0.05.Theconcentration of OCN and OPN in B-F group after 14 d were （0.25 ±0.03） and （0.35 ±0.04） ng/ml,（0.55 ±0.06） ng/ml and （0.59 ±0.06） ng/ml,（0.71 ±0.08） ng/ml and （0.84±0.09） ng/ml,（0.94±0.10） ng/ml and （1.16±0.12） ng/ml,（0.98±0.11） ng/ml and （1.19 ± 0.
作者
黄敏红
梁至洁
梁一丹
彭契六
邓国雄
黄海
池刚毅
徐房添
黎洪棉
Huang Minhong Liang Zhijie Liang Yidan Peng Qiliu Deng Guoxiong Huang Hai Chi Gangyi Xu Fangtian Li Hongmian(Department of Hepatobiliary and Gland Surgery, the Fifth Affiliated Hospital of Guangxi Medical University ＆ Nanning First People＇s Hospital, Nanning 530022, Chin Department of Plastic Surgery, the Fifth Affiliated Hospital of Guangxi Medical University ＆ Nanning First People＇ s Hospital, Nanning 530022, China Medical Inspection Center, the Fifth Affiliated Hospital of Guangxi Medical University ＆ Nanning First People＇ s Hospital, Nanning 530022, China Department of Cardiology, the Fifth Affiliated Hospital of Guangxi Medical University ＆ Nanning First People＇s Hospital, Nanning 530022, Chin Department of Orthopedics, the First Affiliated Hospital of Gannan Medical University, Ganzhou 341000, Chin)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第11期2463-2468,共6页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金（81560316、81560358）
广西自然科学基金重点项目（2016GXNSFDA380016）
广西科学研究与技术开发计划资助项目（桂科攻1598012-1）
南宁市科学研究与技术开发计划资助项目（20153089、zc20153002、zc20153004）
关键词
人参皂苷
乳腺脂肪来源干细胞
细胞增殖
成骨分化
组织工程
Breast adipose derived stem cells
Cell proliferation
Osteogenetic differentiation
Tissue engineering
