姜黄素脂质体对人黄韧带细胞Ⅰ、Ⅲ型胶原mRNA表达的影响

AFFECT OF CURCUMIN LIPOSOMES ON mRNA EXPRESSION OF COLLAGENS TYPE ⅠAND TYPE Ⅲ IN HUMAN LIGAMENTUM FLAVUM CELLS

目的观察姜黄素(Cur)对人黄韧带细胞(LFCs)Ⅰ、Ⅲ型胶原表达的影响,探讨其抗黄韧带纤维化的机制。方法采用组织块加酶预消化法培养LFCs作为研究模型,予以不同浓度姜黄素0、10、50、100、200 ng/ml刺激人LFCs 12 h,采用四甲基偶氮唑盐(MTT)比色法检测姜黄素对人LFCs的抑制效应,采用Real-time PCR法检测姜黄素对人LFCs的Ⅰ、Ⅲ型胶原mRNA表达的影响。结果 MTT检测结果显示:10ng/ml姜黄素作用于人LFCs,表现为增殖倾向,其光吸收值(OD值)显著地高于正常对照组。而在50～100ng/ml的浓度范围内,随着姜黄素药物浓度的增加,其抑制作用逐渐增强,100 ng/ml和200 ng/ml姜黄素能显著抑制人LFCs增殖(p≤0.05);Real-time PCR结果显示200 ng/ml姜黄素组能够显著抑制人LFCsⅠ、Ⅲ型胶原合成(p≤0.05)。结论较高浓度的姜黄素能有效抑制人LFCs增殖和Ⅰ、Ⅲ型胶原合成,对人黄韧带纤维化可能有治疗作用。

Objective To study the affect of curcumin liposomes on expression of the type Ⅰ and Ⅲ collagens of human hgamentum fiavum ceils （LFCs）probing into the mechanism of its anti -fibrosis function. Methods Ligamentum flavum cells were primary cultured with tissue block enzymolytic method. LFCs were exposed to 0,10, 50,100,200 ng/ml curcumin for 12 h. 5 -di -phenytetrazoliumromide （MTT）method was used to analyze the cell proliferation by measuring the optical density（OD） values. Expressions of collagen Ⅰ and Ⅲ mRNA in LFCs were detected by Real - time PCR. Results MTr revealed that curcumin in 10ng/ml showed a LFCs proliferation enhancing trend, and its absorbance value was significantly higher than that in the normal group, but the effect turned to inhibition when concentration increased to over 50 - 200 ng/ml, and became significant inhibition at concentration of 100 and 200 ng/ml（p≤0. 05）. Besides, curcumin also showed markedly inhibition on collagen type Ⅰ and Ⅲ synthesis in LFCs（p≤0. 05）. Conclusion High concentration curcumin can inhibit effectively proliferation and collagen Ⅰ and Ⅲ expression in LFCs, therefore, may has therapeutic effect on the fibrosis of ligamentum flavum in human being.