本文選題：牙齦 + 細胞衰老　； 參考：《上海交通大學學報(醫(yī)學版)》2017年05期

【摘要】：目的·構(gòu)建人牙齦成纖維細胞(hGFs)復制性衰老模型,探討衰老相關生物學改變,為牙周疾病衰老相關研究提供有效的細胞模型。方法·取健康牙齦組織,利用組織塊法體外培養(yǎng)hGFs,免疫熒光染色驗證組織來源;連續(xù)傳代培養(yǎng),計算累積細胞群體倍增指數(shù)(CPD)并繪制曲線;取CPD值為10.82、20.65、29.52、42.22、60.79、70.03的hGFs,CCK-8試劑盒檢測細胞增殖能力改變,real-time PCR檢測衰老相關基因p16~(INK4a)、p21~(Cip1)的表達變化。結(jié)果·連續(xù)傳代培養(yǎng)后,CPD值逐漸增大,達70.03時進入平臺期;隨著CPD值的增大,hGFs細胞形態(tài)呈扁平狀,細胞突起增多;細胞增殖能力減弱;p16~(INK4a)、p21~(Cip1) mRNA表達水平顯著升高(P=0.000)。結(jié)論·連續(xù)傳代培養(yǎng)可構(gòu)建hGFs復制性衰老模型,CPD曲線可反映細胞衰老狀態(tài)。
[Abstract]:Objective to establish a replicating aging model of human gingival fibroblasts (hGFs) and explore the biological changes associated with senescence in order to provide an effective cell model for the study of aging of periodontal diseases.Methods healthy gingival tissue was cultured in vitro with tissue mass method, and the tissue origin was verified by immunofluorescence staining, then the cumulative cell population doubling index (CPD) was calculated and the curves were drawn.The expression of the senescence related gene p16NK4a was detected by real-time PCR with a CPD value of 10.82n 20.655.52 (42.2260.709 / 70.03) and a CCK-8 kit of hGFsfen CCK-8 was used to detect the change of cell proliferative ability and the expression of the senescence related gene p16NK4a (p21 ~ (1)) was detected by real-time PCR.Results after continuous passage culture, the CPD value increased gradually, reaching 70.03 to the plateau stage, with the increase of CPD value, the morphology of hGFs cells was flat, the cell processes increased, and the cell proliferation ability decreased. The expression level of P0.000 mRNA was significantly increased.Conclusion continuous passage culture can construct hGFs replicative senescence model and the curve can reflect the state of cell senescence.
【作者單位】： 上海交通大學醫(yī)學院附屬第九人民醫(yī)院牙周病科上海市口腔醫(yī)學重點實驗室;
【基金】：國家自然科學基金(81570977) 上海市教育委員會高峰高原學科建設計劃~~
【分類號】：R781.4
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本文編號：1731365