AFMK对大鼠牙乳头细胞增殖和分化的影响  

The effects of AFMK on proliferation and differentiation of rat dental papilla cells

作　　者：卢艳红[1] 付深利[1] 何依帆[1] 周红玉[1] 黄芳[1] 刘永亮[1] 高志雄[1] 

机构地区：[1]中山大学光华口腔医学院·附属口腔医院,广东省口腔医学重点实验室,广州510055

出　　处：《中华口腔医学研究杂志（电子版）》2013年第2期16-20,共5页Chinese Journal of Stomatological Research(Electronic Edition)

基　　金：广东省科技计划(2008B030301102);2012年中山大学实验室开放基金(KF201221)

摘　　要：目的探讨N1-乙酰基-N2-甲酰基-5-甲氧基犬尿氨酸(AFMK)对大鼠牙乳头细胞(RDPCs)增殖和分化的影响。方法选用新生SD大鼠,体外分离培养RDPCs。取第4代RDPCs,分别在普通培养条件和矿化诱导条件下,采用MTT比色法检测不同浓度AFMK对RDPCs增殖的影响;矿化诱导条件下,AFMK刺激RDPCs后,PNPP偶氮法、茜素红染色法和免疫组化法分别检测RDPCs的碱性磷酸酶(ALP)活性、矿化基质形成和牙本质涎蛋白的表达。结果在普通培养条件下,加药第2、3、4天后,AFMK低、高浓度组的吸光度(A)值均低于对照组(P<0.05)。在矿化诱导条件下:(1)加药第2、3、4天后,AFMK低、高浓度组的A值均高于对照组(P<0.05);(2)AFMK高浓度组ALP活性高于对照组(P<0.05);(3)AFMK高浓度组茜素红染色A值高于对照组(P<0.05);(4)牙本质涎蛋白染色在对照组呈弱阳性,在AFMK高浓度组呈强阳性。结论普通培养条件下,AFMK抑制RDPCs的增殖;矿化诱导条件下,AFMK则促进其增殖和分化。Objective To investigate the effects of N1-acetyl-N2-formyl-5-methoxy-kynuramine （AFMK） on the proliferation and differentiation of rat dental papilla cells （RDPCs）. Methods Dental papilla cells of neonatal rats were isolated and cultured in vitro. Different concentrations of AFMK were added into the common and osteogenic medium, while the proliferation of RDPCs was examined by MTT assay. ALP activity was measured by analyzing the rate of PNPP hydrolysis. Osteogenie differentiation capacity was determined by alizarin red staining, the expression of dentin sialoprotein （DSP） in RDPCs was detected by immunocytochemistry. Results In common medium, the result of MTr assay showed that the absorbance （A） value of the AFMK groups with various concentration were lower than the control group on day 2, 3 and 4 （P 〈 0.05）. In osteogenic culture induction condition, （1）The result of MTT assay showed that the A value of the AFMK groups with various concentration were higher than the control group on day 2, 3 and 4 （P 〈 0.05）; （2）The ALP aetivaty of high concentration AFMK group was higher than that of the control group （P 〈 0.05）; （3）The alizarin red staining showed that the high concentration AFMK group formed more mineralized matrix than control group （P〈 0.05）; （4）The immunocytochemistry staining showed that positive staining of DSP was found from the high concentration AFMK group compared with control group. Conclusion AFMK inhibitded the proliferation of RDPCs in normal culture condition, whereas promoted proliferation and differentiation in osteogenic culture condition.

关 键 词：N1-乙酰基-N2-甲酰基-5-甲氧基犬尿氨酸 牙乳头细胞 增殖 分化 

分 类 号：R78[医药卫生—口腔医学]