摘要
目的探讨c AMP信号通路对根尖乳头干细胞(SCAP)增殖、迁移的影响。方法酶消化法培养细胞;矿化诱导液和成脂诱导液诱导细胞后,茜素红和油红O染色鉴定细胞多分化潜能;分别用不同浓度的c AMP信号通路激活剂Forskolin和抑制剂H-89处理细胞;MTT和划痕实验检测各组细胞的增殖能力及迁移能力。结果矿化和成脂诱导液分别诱导SCAP后,茜素红和油红O染色显示有钙盐沉积及脂滴形成。低浓度的Forskolin(2.5μmol·L^(-1))促进细胞增殖,高浓度的Forskolin(10μmol·L^(-1),20μmol·L^(-1))则发挥相反作用。用低浓度H-89(5μmol·L^(-1),10μmol·L^(-1))抑制c AMP信号后,SCAP的增殖能力增加,而高浓度H-89(20μmol·L^(-1))在MTT第3天时促进细胞增殖,第5天和第7天时抑制该细胞增殖。迁移结果显示高浓度Forskolin(10μmol·L^(-1),20μmol·L^(-1))抑制SCAP的迁移,而H-89(5μmol·L^(-1),10μmol·L^(-1))促进该细胞的迁移。结论 c AMP信号通路对根尖乳头干细胞增殖、迁移有一定的调节作用。
Objective This study was designed to investigate the influence of the c AMP signaling on proliferation and migration of SCAP. Methods SCAP was cultured by Enzyme digestion method. After mineralization induction and adipogenic induction,the multiple differentiation potential of SCAP was identified by alizarin red staining and oil red O staining. SCAP was exposed to various concentrations of Forskolin and H-89 respectively. Cell proliferation ability and migration ability were checked by MTT assay and wound healing test. Results After mineralization induction and adipogenic induction,calcium deposition and lipid droplet were identified in SCAP by alizarin red staining and oil red O staining. Low concentration of Forskolin( 2. 5 μmol·L^(-1)) promoted proliferation of SCAP,while high concentrations of Forskolin( 10 μmol·L^(-1),20 μmol·L^(-1)) inhibited proliferation of the cell. After inhibiting the signaling by H-89( 5 μmol·L^(-1),10 μmol·L^(-1)),the proliferation of SCAP was increased sharply. 20 μmol·L^(-1)H-89 promoted the proliferation of SCAP at the third day. However the proliferation of SCAP was inhibited at the fifth day and the seventh day by 20 μmol·L^(-1)H-89. The SCAP stimulated by Forskolin( 10 μmol / l,20 μmol·L^(-1)) presented a lower migration capacity. While the SCAP stimulated by H-89( 5 μmol·L^(-1),10 μmol·L^(-1)) exhibited an improved migration capacity. Conclusion The c AMP signaling can influence the proliferation and migration of SCAP.
出处
《安徽医药》
CAS
2016年第5期874-878,共5页
Anhui Medical and Pharmaceutical Journal
基金
安徽省自然科学基金资助项目(No 1408085QH178)
安徽医科大学校科研基金资助项目(No 0401059101)
