胸腺五肽延缓小鼠皮肤成纤维细胞衰老的研究

发布时间：2018-08-05 19:55

【摘要】： 【目的】从细胞水平探讨胸腺五肽延缓衰老的作用和初步机制。【方法】首先对小鼠皮肤成纤维细胞培养方法进行研究,然后通过台盼蓝染色法检测细胞活力、细胞生长曲线测定细胞增殖能力、HE染色法测定细胞形态、染色体分析鉴定其鼠源性、免疫组化染色对成纤维细胞进行特异性鉴定,最后以小鼠皮肤成纤维细胞作为给药载体,以细胞的增殖、传代数、存活天数、细胞内超氧化岐化酶(SOD酶)活力、丙二醛(MDA)含量研究胸腺五肽的抗衰老效应。【结果】在原代小鼠皮肤成纤维细胞培养上,得出一种较常规做法更为快捷、有效的胰蛋白酶消化法,和有利于细胞纯化、生长、传代、冻存的方法,为后续细胞实验奠定基础。通过台盼蓝染色法检测细胞活力,显示原代细胞存活率大于65%、传代细胞存活率大于95%;显示其细胞增殖能力符合一般细胞生长趋势,生长状态稳定;细胞形态测定表明所培养细胞为典型成纤维细胞结构;染色体核型为二倍体n=40,确定了细胞的鼠源性;波形蛋白免疫组化染色成阳性说明其为成纤维细胞。与阴性对照组比较的结果表明,胸腺五肽能明显促进细胞的增殖(P0.001),增加细胞传代数(两代),增加细胞寿命(细胞存活时间从13天延长到了36天),并能促进细胞内总SOD酶活力(P0.001),减少MDA的产生(P0.001)。【结论】胸腺五肽可以促进小鼠皮肤成纤维细胞增殖,增加其传代数,延长其存活时间,并提高细胞内SOD酶活性,减少MDA含量,因此胸腺五肽具有抗衰老作用,同时小鼠皮肤成纤维细胞模型有可能成为一种高效率的抗衰老细胞模型。
[Abstract]:[objective] to investigate the effect and mechanism of thymopentapine on delaying senescence from cell level. [methods] the culture method of mouse skin fibroblasts was studied firstly, and then the viability of mouse skin fibroblasts was detected by trypan blue staining. Cell growth curve and HE staining were used to determine the cell morphology, the mouse origin was identified by chromosome analysis, and the fibroblasts were identified by immunohistochemical staining. Finally, mouse skin fibroblasts were used as drug delivery carriers. The antiaging effects of thymosin on primary mouse skin fibroblasts were studied by cell proliferation, transmission algebra, survival days, intracellular superoxide dismutase (SOD) activity and malondialdehyde (MDA) content. A method of trypsin digestion, which is faster and more effective than the conventional method, is obtained, which is beneficial to cell purification, growth, passage and cryopreservation, and lays a foundation for subsequent cell experiments. The viability of the cells was detected by trypan blue staining, which showed that the survival rate of the primary cells was greater than 65 and the survival rate of the passage cells was greater than 95%, which showed that the proliferation ability of the cells was consistent with the general cell growth trend and the growth state was stable. The morphological analysis showed that the cultured cells were typical fibroblast structure, the chromosome karyotype was diploid nong40, which confirmed the murine origin of the cells, and the positive staining of vimentin showed that the cells were fibroblasts. Compared with the negative control group, the results showed that, Thymopeptide could significantly promote cell proliferation (P0.001), increase cell transmission algebra (two generations), increase cell life (cell survival time from 13 days to 36 days), promote the total intracellular SOD enzyme activity (P0.001), and reduce the production of MDA (P0.001). [ Conclusion Thymosin can promote the proliferation of mouse skin fibroblasts. The thymic pentapeptide has anti-aging effect because it increases its transmission algebra, prolongs its survival time, increases the activity of SOD enzyme in cells and reduces the content of MDA. At the same time, the mouse skin fibroblast model may become a high-efficiency anti-aging cell model.
【学位授予单位】：天津大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R392

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