【作者】 毛家奇； 李淑娟； 胡永青； 杨冬茹；

【Author】 MAO Jiaqi;LI Shujuan;HU Yongqing;YANG Dongru;Stomatological Hospital of Hebei Medical University;

【机构】 河北医科大学口腔医院；

【摘要】 目的 :研究炎症对人牙周膜干细胞成骨分化3、7和14天自噬相关蛋白和mRNA表达的影响。方法 :培养原代人牙周膜干细胞,在正常和炎症来源的牙周膜干细胞成骨分化3、7和14天,Realtime PCR检测ALP、LC3B、Beclin1mRNA的表达,Western Blot检测ALP、LC3B、Beclin1的蛋白表达水平,采用SPSS22.0软件包对数据进行统计学分析。结果 :1、成骨诱导分化3、7和14天、炎症来源的牙周膜干细胞ALP mRNA的表达水平均明显低于正常来源的牙周膜干细胞(P<0.05),炎症来源的牙周膜干细胞ALP蛋白表达水平明显低于正常来源的牙周膜干细胞(P<0.05)。2、RT-PCR结果显示,成骨诱导分化3天,炎症来源的牙周膜干细胞LC3B、Beclin1 mRNA的表达水平均低于正常来源的牙周膜干细胞,但无显著统计学差异(P>0.05),成骨诱导分化7和14天,炎症来源的牙周膜干细胞LC3B、Beclin1 mRNA的表达水平明显低于正常来源的牙周膜干细胞(P<0.05)。3、Western Blot结果显示,成骨诱导分化3天,炎症来源的牙周膜干细胞LC3B、Beclin1蛋白表达水平均低于正常来源的牙周膜干细胞,但无显著统计学差异(P>0.05),成骨诱导分化7和14天,炎症来源的牙周膜干细胞LC3B、Beclin蛋白表达水平明显低于正常来源的牙周膜干细胞(P<0.05)。结论 :炎症对牙周膜干细胞成骨分化3天自噬表达水平无明显影响,炎症明显降低了牙周膜干细胞成骨分化7和14天自噬表达水平。更多还原

【Abstract】 Objective: To study the effect of inflammation on the expression of autophagy related proteins and mRNA in human periodontal ligament stem cells at 3, 7 and 14 days. Methods: primary human periodontal ligament stem cells were cultured and identified. On the 3rd, 7th and 14th day of osteogenic differentiation of normal and inflammatory periodontal ligament stem cells, real time PCR was used to detect the expression of ALP, lc3b, Beclin1 mRNA, Western blot was used to detect the protein expression of ALP, lc3b, Beclin1, and spss22.0 was used to analyze the data. Results: 1. The expression levels of ALP mRNA in inflammatory periodontal ligament stem cells were significantly lower than those in normal periodontal ligament stem cells(P < 0.05), and the expression levels of ALP protein in inflammatory periodontal ligament stem cells were significantly lower than those in normal periodontal ligament stem cells(P < 0.05). 2. The results of RT-PCR showed that the expression levels of lc3b, Beclin 1 mRNA in inflammatory periodontal ligament stem cells were lower than those in normal periodontal ligament stem cells, but there was no significant difference(P > 0.05) Peripheral membrane stem cells(P < 0.05). 3. Western blot showed that the expression levels of lc3b, Beclin 1 protein in inflammatory periodontal ligament stem cells were lower than those in normal periodontal ligament stem cells, but there was no significant difference(P > 0.05). The expression levels of lc3b, Beclin 1 protein in inflammatory periodontal ligament stem cells were lower than those in normal periodontal ligament stem cells at 7 and 14 days after osteogenesis Periodontal ligament stem cells(P < 0.05). Conclusions: Inflammation had no significant effect on the autophagy expression level of periodontal ligament stem cells on the 3rd day, but decreased the autophagy expression level of periodontal ligament stem cells on the 7th and 14th day.更多还原