黄瓜籽正丁醇提取物对人牙髓干细胞增殖及成牙向分化的影响

Effects of N-butanol Extract of Cucumissativus Seeds on Proliferation and Odontogenic Differentiation of Human Dental Pulp Stem Cells

目的:探讨黄瓜籽正丁醇提取物(n-butanol extract of cucumis sativus seeds,CSB)对人牙髓干细胞(human dental pulp stem cells,hDPSCs)增殖及成牙向分化能力的影响。方法:体外扩增培养hDPSCs,采用细胞计数法(cell counting kit-8,CCK-8)法检测CSB对hDPSCs增殖能力的影响;通过碱性磷酸酶(ALP)染色及活性测定、Western blot检测牙本质涎磷蛋白(dentin sialophosphoprotein,DSPP)和Ⅰ型胶原(collagenⅠ,COLⅠ)等矿化相关蛋白的表达水平观察细胞成牙向分化能力的变化。结果:CCK-8结果显示,与对照组相比,100mg/L CSB可显著促进hDPSCs的增殖(P<0.05),500mg/L组则明显抑制hDPSCs的增殖(P<0.05);经矿化诱导后,100mg/L CSB组ALP染色明显加深,活性显著升高(P<0.05);Western blot结果显示100mg/L CSB组DSPP和COLⅠ的表达均高于对照组(P<0.05)。结论:适宜浓度的CSB可促进hDPSCs的增殖及成牙向分化。

Objective：To investigate the effects of n-butanol extract of Cucumis sativus seeds（CSB）on proliferation and odontogenic differentiation of human dental pulp stem cells（hDPSCs）.Methods：HDPSCs were isolated and exposed to different concentrations of CSB solutions.The proliferation ability was evaluated by Cell Counting Kit-8（CCK-8）assay.The odontogentic potential of hDPSCs was assessed by alkaline phosphatase（ALP）staining and activity assay.In addition,Western blot analysis was conducted to identify the expressions of dentin sialophosphoprotein（DSPP）and Collagen I（COL I）which were related proteins on odontogenic differentiation of hDPSCs.Results：CCK-8 assay showed that CSB significantly enhanced the proliferation of hDPSCs at a concentration of 100 mg/L,while CSB remarkably suppressed the growth of hDPSCs at a higher concentration of 500 mg/L.The hDPSCs cultured with 100 mg/L CSB obviously induced positive ALP staining and had higher ALP activity compared with the control group.Furthermore,Western blot analysis demonstrated that the expression levels of odontogenic proteins DSPP and COL I were significantly up-regulated in 100 mg/L CSB-treated hDPSCs compared with the control group.Conclusion：The results indicate that an appropriate concentration of CSB could enhance the proliferation and odontogenic differentiation of hDPSCs.