LMK-235对牙周膜细胞成骨和成牙本质分化的影响

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 ligament cells，hPDLCs）早期成骨和成牙本质分化的影响。方法通过酶消化法获得hPDLCs，分别用浓度为0、50、100、250、500 nmol/L的LMK?235处理第三代hPDLCs 3 d。MTT法检测hPDLCs的增殖，同时qRT?PCR检测成骨及成牙本质相关因子Runx2、ALP及DMP?1 mRNA的表达水平。结果 MTT结果显示100 nmol/L的LMK?235对hPDLCs增殖具有促进作用。qRT?PCR结果表明100 nmol/L处理组Runx2 mRNA的表达水平为对照组的1.77倍（P＜0.05）；而ALP mRNA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 mRNA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hPDLCs增殖，并通过上调Runx2、ALP、DMP?1等成骨及成牙本质相关因子mRNA表达来促进hPDLCs早期成骨及成牙本质分化。

Effects of LMK-235 on osteoblast/odontoblast differentiation in hPDLCs

Objective To investigate the effects of typeⅡa histone deacetylase inhibitor LMK?235 during early os?teoblast/odontoblast differentiation in hPDLCs. Methods hPDLCs were obtained by the collagenase digestion method. hPDLCs at the 3rd passage 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 hPDLCs was tested by MTT and qRT?PCR was used to detect mRNA expression levels of Runx2, ALP and DMP?1 3 d later. Results MTT assay showed that cell proliferation in hPDLCs treated with 100 nmol/L LMK?235 was increased significantly compared with the con?trol group (P<0.05). The expression of Runx2 mRNA in the 100 nmol/L group was 1.77 times of the control groups (P<0.05). The expressions of ALP mRNA 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 mRNA in the 50 and 100 nmol/L groups were higher than the control groups (P<0.05). Conclusion TypeⅡa histone deacetylase inhibitor LMK?235 could accelerate cell proliferation in hPDLCs at the concentration of 100 nmol/L, and regulate early osteoblast/odontoblast differentiation by upregulating the mRNA expressions of Runx2, ALP and DMP?1.