柚皮苷对人牙周膜细胞功能调节的作用

Experimental study on the functional regulation of naringin in human periodontal ligament cells

目的:通过研究中药有效成分柚皮苷对人牙周膜细胞(hPDLCs)增殖、矿化成骨及骨保护素(OPG)表达的影响,探讨柚皮苷对人牙周膜细胞增殖及成骨功能的调节作用。方法:采用酶消化结合组织块法,体外原代培养hPDLCs,通过四甲基偶氮唑蓝(MTT)比色法、酶联免疫吸附法和半定量反转录聚合酶链反应(RT-PCR)等方法,测定不同质量浓度(100、10、1.0、0.1、0.01mg/L)柚皮苷作用后,hPDLCs增殖、碱性磷酸酶(ALP)活性、Ⅰ型胶原蛋白、OPG表达的变化,采用SPSS16.0统计包对数据进行统计学处理。结果:原代培养的hPDLCs形态良好,1.0mg/L浓度组的柚皮苷对hPDLCs增殖、ALP活性和Ⅰ型胶原蛋白表达的促进作用最强,此浓度柚皮苷对细胞OPG mRNA的调节作用在测定时段内呈时间依赖性。结论:柚皮苷有促进hPDLCs增殖及向成骨方向转化的作用。

PURPOSE：To detect the effect of naringin on human periodontal ligament cells＇（hPDLCs） proliferation,bone formation and OPG mRNA expression.METHODS： hPDLCs were primarily cultured and identified in vitro through enzyme digestion combined tissue culture method.With 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT）method,enzyme linked immunosorbent assay and RT-PCR,the hPDLCs＇ proliferation,alkaline phosphatase（ALP）activity,expression of collagen protein-Ⅰand OPG mRNA were observed after treatment with different concentrations（100,10,1.0,0.1,0.01mg/L）of naringin at different times.SPSS16.0 software package was used for statistical analysis.RESULTS： Primary cultured hPDLCs had good shape;Significant promotion of proliferation,ALP activity and collagen protein-Ⅰexpression of hPDLCs with naringin was found at the dose of 1.0 mg/L;and 1.0mg/L naringin regulated the expression of OPG mRNA in time-dependent manner.CONCLUSION： Naringin might significantly promote the proliferation of hPDLCs and conversion into the osteoblast.Supported by National Natural Science Foundation of China（30940092） and ＂13115＂ Major Scientific and Technological Innovation Special Projects of Shanxi Province（2009ZDKG-77）.