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高通量测序解析驴骨胶原蛋白促进伤口愈合的潜在分子机制
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作者 廖峰 樊雨梅 +2 位作者 帖航 赵海晴 苏宁 《食品工业科技》 CAS 北大核心 2022年第6期358-364,共7页
探讨驴骨胶原蛋白对成纤维细胞(NHDFs细胞)增殖、迁移和Ⅰ型胶原蛋白分泌的影响以及可能的作用通路。通过八肽胆囊收缩素(CCK-8)法、5-溴脱氧尿嘧啶核苷(BrdU)法和划痕实验,研究驴骨胶蛋白对成纤维细胞增殖和迁移的影响,并利用转录组测... 探讨驴骨胶原蛋白对成纤维细胞(NHDFs细胞)增殖、迁移和Ⅰ型胶原蛋白分泌的影响以及可能的作用通路。通过八肽胆囊收缩素(CCK-8)法、5-溴脱氧尿嘧啶核苷(BrdU)法和划痕实验,研究驴骨胶蛋白对成纤维细胞增殖和迁移的影响,并利用转录组测序技术(RNA-seq)分析驴骨胶原蛋白作用于成纤维细胞的差异表达基因。结果表明,驴骨胶原蛋白对NHDFs细胞无毒性,且在0.16~5.0 g/mL内对NHDFs细胞有一定的促生长作用。0.75~1.50 g/mL的驴骨胶原蛋白能够促进NHDFs细胞增殖,且驴骨胶原蛋白对NHDFs细胞增殖的影响呈现剂量依赖关系。1.50 g/mL的驴骨胶原蛋白可显著促进NHDFs细胞迁移和Ⅰ胶原蛋白的分泌。驴骨胶原蛋白作用于成纤维细胞后,共筛选出384个差异表达基因,其中172个基因上调,212个基因下调。GO功能富集分析结果表明,差异表达基因与中性粒细胞趋化性、中性粒细胞迁移、粒细胞趋化性等有关;KEGG通路富集分析表明,差异基因参与细胞因子-细胞因子受体相互作用、化学致癌、IL-17和TNF等重要生物学通路。推测,驴骨胶原蛋白可能通过IL-17和TNF-α介导CXCL趋化因子发挥促进皮肤伤口愈合的作用。本文明确了驴骨胶原蛋白促进伤口愈合的效果,填补驴骨胶原蛋白促进伤口愈合的研究空白,为驴骨胶原蛋白高值转化提供了思路。 展开更多
关键词 驴骨胶原蛋白 伤口愈合 nhdfs细胞 Ⅰ型胶原蛋白 RNA-seq技术
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Induction of Apoptosis in the Human Prostate Cancer Cell Line DU-145 by a Novel Micronutrient Formulation
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作者 M. Waheed Roomi Neha Shanker +1 位作者 Aleksandra Niedzwiecki Matthias Rath 《Open Journal of Apoptosis》 2015年第1期11-21,共11页
Prostate cancer, the most frequently diagnosed cancer in men, primarily affects males aged 55 and older and is more common in African Americans than Caucasians. Once the cancer has metastasized, current treatments are... Prostate cancer, the most frequently diagnosed cancer in men, primarily affects males aged 55 and older and is more common in African Americans than Caucasians. Once the cancer has metastasized, current treatments are generally ineffective. We have identified a novel anti-neoplastic agent, a specifically designed nutrient mixture (NM), containing ascorbic acid, lysine, proline and green tea extract that demonstrates a broad spectrum of anti-tumor activity against a number of human cancer cell lines. In a previous study NM significantly inhibited prostate tumor in nude mice. In this study, we tested whether the formulation exerts its anti-tumor effects through induction of apoptosis on prostate cancer cell line DU-145. The effect of the nutrient mixture (NM) on cell growth inhibition in DU-145 cells was examined by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Morphological changes and caspase activation associated with apoptosis induction was checked by H&E staining and Live Green Caspase assay, respectively. The NM was found to be slightly toxic to DU-145 cells at 100 μg/ml, but significantly toxic at 500 μg/ml and 1000 μg/ml. Percentage of cells undergoing apoptosis also increased from 6% at 100 μg/ml to 49% at 500 μg/ml and 83% at 1000 μg/ml, with greater number of cells showing morphological changes such as condensed nuclei and an acidophilic cytoplasm at higher concentrations. For the purpose of comparison, NM was also tested on a normal human dermal fibroblast (NHDF) cell line which exhibited far less apoptosis induction as compared to DU-145 cells. The percentage of cells undergoing apoptosis in case of NHDF cells was 7% at 100 μg/ml, 25.6% at 500 μg/ml and 76.5% at 1000 μg/ml. Our results demonstrate that the NM is effective in inhibiting cancer cell viability and inducing apoptosis in prostate cancer DU-145 cells and can thus be used as an effective treatment for prostate cancer. 展开更多
关键词 PROSTATE Cancer DU-145 NHDF APOPTOSIS CYTOTOXICITY CASPASE MTT
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