| 聂爱婷,马 波,胡利平,张秀峰,饶 旼,聂胜洁.衰老程度评价生物学指标在人胚肺二倍体细胞的筛选研究[J].,2017,17(17):3255-3258 |
| 衰老程度评价生物学指标在人胚肺二倍体细胞的筛选研究 |
| The Screening Study of Biological Indicators for Evaluation of the Senescence in Human Embryonic Lung Diploid Cells |
| 投稿时间：2016-07-19 修订日期：2016-08-15 |
| DOI：10.13241/j.cnki.pmb.2017.17.013 |
| 中文关键词: 细胞衰老 代龄 细胞形态 β-半乳糖苷酶 衰老评价 |
| 英文关键词: Cellular senescence Generation aging Cell morphology β-galactosidase enzyme Senescence evaluation |
| 基金项目:国家科技部863计划项目（2014AA021002） |
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| 中文摘要: |
| 摘要 目的：通过在微观结构水平和分子水平检测不同代次人胚肺二倍体细胞衰老相关的指标值，筛选出能够系统性评价人胚肺二倍体细胞衰老程度的生物学指标。方法：将Walvax-2细胞分成16个代次，在倒置显微镜下观察不同代次Walvax-2细胞生长情况及形态学变化；在透射电镜下观察不同代次Walvax-2细胞生长情况和衰老情况；通过检测SA-β-gal染色阳性率，分析不同代次Walvax-2细胞β-半乳糖苷酶的表达水平。结果：细胞可传58代。倒置显微镜下：随细胞代次的增加，可见颜色逐渐加深，色素积累；透射电子显微镜观察到：P20到P43细胞正常且骨架结构明显，细胞核完整，具有较高的核质比,P44和P45开始出现衰老现象，细胞质色素积聚加深，核膜不同程度内折，P50和P55细胞衰老更加明显；β-半乳糖苷酶的阳性染色率和细胞代龄之间呈显著相关(P <0.01)。结论：细胞形态学变化，β-半乳糖苷酶阳性率变化与Walvax-2细胞代龄增长显著相关,可选择作为Walvax-2细胞衰老程度评价的生物学指标。 |
| 英文摘要: |
| ABSTRACT Objective: To collect the biological indicators of state closely related to cell growth and cell aging, through the mi- crostructure level and molecular level testing parameter values which aging related to human embryo lung diploid cells of different gener- ation times, Screening the biological indicators which can systemic evaluation of aging degree of human embryo lung diploid cell. Methods: The Walvax-2 cells were split into 16 generations. The growth and morphological of Walvax - 2 cells from different generations were ob- served under inverted microscope. Cells from different generations were observed by transmission electron microscope. To analysis the expression level of β-galactosidase enzyme of different generations Walvax - 2 cells by detecting the rate of SA -β- gal staining positive cells. Results: Cells can grow to 58 generations, with the increase of cell generation time, visible color deepened gradually, pigment accu- mulation by the inverted microscope. The transmission electron microscope observed: cell morphology frame structure, and nucleus are normal and intact from P20 to P43, has the high nuclear mass ratio. Senescence cells P44 and P45 start to appear aging phenomenon. The consequences of senescence cells include irregularly shaped nucleus, nuclear membrane in different level fold, nucleolus decrescent, con- tents increased, endoplasm retinal cavity expansion, the decrease in the number of ribosomes, cytoplasmic pigment deepen and free bub- ble formation, cell autophagy in cell. Senescence cells more evident in P50 and P55. β-galactosidase enzyme staining positive rate and cell generation is significant correlation (P<0.01). Conclusion: Cell morphology and the positive activity of β-galactosidase enzyme were significantly associated with generation growth, it can act as a biological indicator to evaluate the degree of senescence of Walvax-2 cell. |
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