荜茇提取物对大鼠骨髓间质干细胞MSC的增殖作用及与化学官能团的关系

目的:观察荜茇挥发油和水溶性部分对大鼠骨髓间质干细胞(mesenchymal stem cells,MSC)体外增殖的影响,并进一步探讨了与分子中化学官能团的关系.方法:使用密度梯度法分离大鼠骨髓间质干细胞,体外培养,经Brdu标记和CD44染色及两者双重染色鉴定后,观察在培养液中添加不同浓度荜茇挥发油及其水溶性部分的条件下MSC的生长变化.应用形态学、MTT法和5-溴脱氧尿嘧啶(5-Bromodeoxyuridine,Brdu)、增殖细胞核抗原(Proliferation cellnuclear antigen,PCNA)免疫细胞化学方法测定细胞增殖活性.结果:荜茇挥发油以浓度依赖方式促进MSC增殖活力和增加Brdu、PCNA阳性细胞数,与对照组比较有显著性(P＜0.05).经GC-MS检测,荜茇挥发油含C=C(50.66%)、-OH(27.02%)、多种官能团(5.05%)等;而荜茇水提物对MSC无增殖作用.结论:荜茇挥发油明显促进MSC的增殖,这种作用可能与分子中C=C、-OH等官能团有关.

Study on Proliferation Effect of extracts of Piper longum on Mesenchymal Stem Cells of Rat Bone Marrow and the Relationship to Chemical Functional Groups

Objective: To observe the effect of volatile oil and aqueous soluable part of Piper longum on proliferation of rat mesenchymal stem cell(MSC)and the chemical functional groups. Methods: Mesenchymal stem cells were dissociated from rat bone marrow and marked by Brdu, and the expression of CD44, CD54 and double label of Brdu and CD44. The growth of rat mesenchymal stem cell under volatile oil and aqueous soluable part of Piper longum was observed by means of cell viability measurement(MTT)and morphological observation and Brdu, PCNA immunohistochemical methods. Results: Volatile oil of Piper longum could promote the cell viability of MSC and the number of Brdu, PCNA positive cell in dose-dependant. There was significant difference in comparision with control groups, C=C(50.66%),-OH(27.02%)and other functional groups in volatile oil of Piper longum were determined by GC-MS, but the aqueous soluable part of Piper longum could not promote the proliferation of MSC. Conclusions: Volatile oil of Piper longum which consists of C=C, -OH, and other functional groups has strong effect on enhancing proliferation of MSC.