文章摘要
黄鉴栎,史 凡,章非敏,王长青.姜黄素促进大鼠骨髓间充质干细胞的体外增殖及成骨分化[J].南京医科大学学报,2020,(12):1868~1873
姜黄素促进大鼠骨髓间充质干细胞的体外增殖及成骨分化
Curcumin promoted proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells in vitro
投稿时间:2020-11-03  
DOI:10.7655/NYDXBNS20201226
中文关键词: 姜黄素  干细胞  骨组织工程  成骨分化
英文关键词: curcumin  stem cells  bone tissue engineering  osteogenesis differentiation
基金项目:国家重点研发计划纳米科技重点专项(2016YFA0201704);江苏省高校优势学科建设工资资助项目(2018?87)
作者单位
黄鉴栎 南京医科大学口腔疾病研究江苏省重点实验室南京医科大学附属口腔医院修复科江苏 南京 210029 
史 凡 南京医科大学口腔疾病研究江苏省重点实验室南京医科大学附属口腔医院修复科江苏 南京 210029 
章非敏 南京医科大学口腔疾病研究江苏省重点实验室南京医科大学附属口腔医院修复科江苏 南京 210029 
王长青 南京医科大学江苏 南京 211166 
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中文摘要:
      目的:研究姜黄素对大鼠骨髓间充质干细胞(rat bone marrow mesenchymal stem cell,rBMSC)生物学行为的影响。方法:将含不同浓度梯度姜黄素溶液的完全培养基与rBMSC共培养,通过CCK?8法检测细胞增殖情况,确定姜黄素溶液的最适浓度;将rBMSC与4 μg/mL姜黄素共孵育,0 μg/mL姜黄素处理组作为对照,通过碱性磷酸酶(alkaline phosphatase,ALP)染色和活性检测评估其早期成骨分化水平;使用茜素红染色评估成骨分化晚期细胞外基质矿化情况;利用实时荧光定量逆转录PCR检测细胞成骨诱导7、14 d时成骨指标骨形态发生蛋白(bone morphogenetic protein?2,Bmp2)、核心结合因子α1(core binding factor alphal α1,Runx2)、骨钙素(osteocalcin,Ocn)、骨桥蛋白(osteopontin,Opn)、成骨细胞特异基因(osterix,Osx)的表达。结果:CCK?8结果显示含不同浓度姜黄素溶液的培养基中的rBMSC均能持续增殖,4 μg/mL姜黄素组对rBMSC增殖的促进作用最为显著(与对照组相比,P < 0.001);ALP活性检测及染色结果显示姜黄素组细胞的ALP活性高于对照组(P < 0.001);茜素红染色结果显示姜黄素组的细胞外基质矿化水平高于对照组;实时荧光定量逆转录PCR结果显示姜黄素组细胞的相关成骨基因Bmp2、Runx2、Ocn、Opn、Osx的表达均高于对照组(P < 0.05)。结论:姜黄素能够促进rBMSC的体外增殖及成骨分化。
英文摘要:
      Objective:This study aims to observe the effect of curcumin on the biological behavior of rat bone marrow mesenchymal stem cells(rBMSC). Methods:Complete medium containing curcumin with different concentration gradient was co?cultured with rBMSC. Cell proliferation was detected by CCK?8 method to determine the optimal concentration of curcumin solution. rBMSC were co?incubated with 4 μg/mL curcumin and treated with 0 μg/mL curcumin as control group. The early osteogenic differentiation of rBMSC was evaluated by ALP staining and activity detection. Alizarin red staining was used to evaluate the mineralization of extracellular matrix in late osteoblast differentiation. Real?time fluorescence quantitative reverse transcription PCR was used to detect the expression of bone morphogenetic protein?2(Bmp2),core binding factor alphal 1(Runx2),osteocalcin(Ocn),osteopontin(Opn),and osteo?specific gene(Osterix,Osx)after 7 and 14 days of osteogenesis induction. Results:CCK?8 results showed that rBMSC in culture medium containing curcumin of different concentrations could proliferate continuously,and the promotion effect of 4 μg/mL curcumin on rBMSC was the most significant(compared with the control group,P < 0.001). ALP activity in curcumin group was higher than that in control group(P < 0.001). The results of alizarin red staining showed that the extracellular matrix level of curcumin group was higher than that of the control group. The expression of Bmp2,Runx2,Ocn,Opn and Osx in the curcumin group was higher than that in the control group(P < 0.05). Conclusion:Curcumin can promote the proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells in vitro.
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