松花粉抗成纤维细胞复制性衰老的机制.docVIP

松花粉抗成纤维细胞复制性衰老的机制 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：松花粉抗成纤维细胞复制性衰老的机制 1 1 材料与方法 3 2 结 果 5 3 讨 论 6 文2：辐射诱导细胞衰老的机制研究进展 9 1 p53-p21信号通路 10 2 p16-RB信号通路 11 3 miRNAs参与辐射诱导细胞衰老的调控 12 4 总结与展望 13 参考文摘引言： 14 原创性声明（模板） 15 文章致谢（模板） 16 正文 松花粉抗成纤维细胞复制性衰老的机制 文1：松花粉抗成纤维细胞复制性衰老的机制 【Abstract】 Objective To observe the effect of pine pollen on the surface area of single cell, the positive rate of senescenceassociated (SA)βgalactosidase stain, the expressio of p16INK4A and p21CIP1 and the phase of cell cycle in 2BS cells. Methods The model of aging cell were made by 2BS cells. The surface area of single cell in each groups were analyzed by the method of biological stereology, the positive rate of SAβgalactosidase stain was determined by immunohistochemistry，the phase of cell cycle was measured by flow cytometry, and the expressio of p16INK4A,p21CIP1mRNA were detected by RTPCR. Results The 56PD cell showed the typical shape changes of aging, the positive rate of SAβgal and the rate of the phase of G1 were both greatly increased (P). After the treatment of pine pollen, the surface area of single cell, positive rate of SAβgal and the rate of the phase of G1 were significantly decreased (P), the expressio of p16INK4A and p21CIP1mRNA were both significantly downregulated (P). Conclusio Pine pollen has the antiaging effect on aging 2BS cells, and its probable molecular mechanism may be related with downregulating the expressio of p16INK4A and p21CIP1mRNA and then improving the G1 phase blocking. 【Key words】 Cell senescence;Pine pollen;Biological stereology;Cell cycle;p16;p21 诱发细胞衰老的主要刺激信号分为端粒和非端粒信号。细胞衰老受多组基因(如p19,p18,p14,MAPK,MEK，Ras,Raf等)调控。细胞衰老的信号通路因细胞种类而异，在人胚肺二倍体成纤维(2BS)细胞中这两大类刺激信号主要诱导p53p21pRb和p16pRb途径。其中，p16与p21是两条信号途径中的核心调控位点，也是目前的研究热点。松花粉能提高衰老小鼠体内抗氧化酶的活性，抑制自由基的氧化损伤〔1〕;降低线粒体mtDNA4977在衰老细胞的突变率;提高衰老细胞端粒酶活性〔2〕。本实验以衰老细胞的单细胞面积、β半乳糖苷酶染色(SAβgal)阳性率及细胞周期等指标对衰老细胞模型进行检测，选取p16及p21作为细胞衰老的分子检测指标，探讨松花粉通过作用于细胞内在基因调控机制抗衰老的可能性。 1 材料与方法 细