LMK-235对牙周膜细胞成骨和成牙本质分化的影响

Effects of LMK-235 on osteoblast/odontoblast differentiation in h PDLCs

目的探讨Ⅱa类组蛋白去乙酰化酶抑制剂LMK-235对人牙周膜细胞(human periodontal ligamentcells,h PDLCs)早期成骨和成牙本质分化的影响。方法通过酶消化法获得h PDLCs,分别用浓度为0、50、100、250、500 nmol/L的LMK-235处理第三代h PDLCs 3 d。MTT法检测h PDLCs的增殖,同时q RT-PCR检测成骨及成牙本质相关因子Runx2、ALP及DMP-1 m RNA的表达水平。结果 MTT结果显示100 nmol/L的LMK-235对h PDLCs增殖具有促进作用。q RT-PCR结果表明100 nmol/L处理组Runx2 m RNA的表达水平为对照组的1.77倍(P<0.05);而ALP m RNA的表达水平在实验组均高于对照组(P<0.05),同时100 nmol/L处理组表达量最高;DMP-1 m RNA的表达水平在50 nmol/L及100 nmol/L组较对照组升高(P<0.05)。结论浓度为100 nmol/L的Ⅱa类组蛋白去乙酰化酶抑制剂LMK-235促进h PDLCs增殖,并通过上调Runx2、ALP、DMP-1等成骨及成牙本质相关因子m RNA表达来促进h PDLCs早期成骨及成牙本质分化。

Objective To investigate the effects of type Ⅱa histone deacetylase inhibitor LMK-235 during early os-teoblast/odontoblast differentiation in h PDLCs. Methods h PDLCs were obtained by the collagenase digestion method.h PDLCs at the 3rdpassage were treated with medium containing 10% fetal bovine serum mixed with different concentra-tions of LMK-235（0, 50, 100, 250, 500 nmol/L）, respectively. Proliferative capability of h PDLCs was tested by MTT andq RT-PCR was used to detect m RNA expression levels of Runx2, ALP and DMP-1 3 d later. Results MTT assay showedthat cell proliferation in h PDLCs treated with 100 nmol/L LMK-235 was increased significantly compared with the con-trol group（P〈0.05）. The expression of Runx2 m RNA in the 100 nmol/L group was 1.77 times of the control groups（P〈0.05）. The expressions of ALP m RNA in all the experimental groups were significantly higher than that in control groups（P〈0.05）, and the expression in the 100 nmol/L groups was the highest. The expressions of DMP-1 m RNA in the 50 and100 nmol/L groups were higher than the control groups（P〈0.05）. Conclusion Type Ⅱa histone deacetylase inhibitorLMK-235 could accelerate cell proliferation in h PDLCs at the concentration of 100 nmol/L, and regulate early osteoblast/odontoblast differentiation by upregulating the m RNA expressions of Runx2, ALP and DMP-1.