| 黄敬文,王 景,安丽凤,段剑飞,代 宇,高宏伟.地龙蛋白对人增生性瘢痕细胞增殖抑制作用机制研究[J].,2017,17(35):6828-6832 |
| 地龙蛋白对人增生性瘢痕细胞增殖抑制作用机制研究 |
| Earthworm Protein of Hyperplastic Scar Cell Proliferation Inhibition Mechanism Research |
| 投稿时间：2017-08-16 修订日期：2017-09-10 |
| DOI：10.13241/j.cnki.pmb.2017.35.007 |
| 中文关键词: 地龙 HSF细胞 Cyclin D1 Caspase-3 TGF-β1 Bcl-2 Mcl-2 |
| 英文关键词: Dragon HSF cells Cyclin D1 Caspase-3 TGF-β1 Bcl-2 Mcl-2 |
| 基金项目:黑龙江省自然科学基金面上项目（H2015023）；黑龙江省博士后管理站课题（LBH-Z13198）；黑龙江中医药大学博士创新基金项目（2013bs03） |
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| 中文摘要: |
| 摘要 目的：研究地龙蛋白对人增生性瘢痕细胞增殖抑制作用机制。方法：培养人增生性瘢痕成纤维细胞(HSF)，将传代好的HSF细胞分为四组，分别加入地龙样品浓度为0 mg/mL（对照组）、0.3125 mg/mL（低剂量组）、0.625 mg/mL（中剂量组）、1.25 mg/mL（高剂量组）。应用MTS法检测细胞活力，流式细胞仪检测细胞凋亡率，RT-PCR法和Western -blot法测定Cyclin D1、Caspase-3、TGF-β1的表达，Western -blot法测定Bcl-2、Mcl-2的表达。结果：地龙蛋白对HSF细胞的增殖活力有抑制作用。浓度0.625 mg/mL、1.25 mg/mL样品组作用细胞48 h后凋亡率为分别为31.5 %和69.2 %，（P<0.01）；RT-PCR显示，浓度0.625 mg/mL、1.25 mg/mL样品组能够增加Caspase-3的基因表达，降低CyclinD1的基因表达，降低TGF-β1的基因表达，（P<0.01）；浓度0.3125 mg/mL样品组与空白对照组比较有显著性差异（P<0.05）。W-B方法显示，各样品组能够增加Caspase-3的蛋白表达量，降低Cy- clinD 1的蛋白表达量，降低TGF-β1的蛋白表达量，（P<0.01）；浓度0.625 mg/mL、1.25 mg/mL样品组能够降低Bcl-2蛋白表达量，（P<0.01）；浓度0.3125 mg/mL样品组Bcl-2的蛋白表达量与空白对照组比较有显著性差异（P<0.05）。结论：地龙蛋白诱导细胞死亡，作用机制与下调Cyclin D1的基因和蛋白表达，上调Caspase-3的基因和蛋白表达，下调TGF-β1的基因和蛋白表达，下调Bcl-2蛋白表达量有关。 |
| 英文摘要: |
| ABSTRACT Objective: The mechanism of the proliferation inhibition of hypertrophic scar cells was studied. Methods: Cultured hy- perplastic scar fibroblasts(HSF), HSF cells were divided into four groups after generation, add earthworm sample separately, the concen- tration is 0 mg/mL(The control group), 0.3125 mg/mL(Low dose group), 0.625 mg/mL (Middle dose group), 1.25 mg/mL (High dose group). Cell vitality was detected by MTS method. Cell apoptosis rate was detected by flow cytometry. The expression of Cyclin D1, Caspase - 3 and TGF-β1 was determined by Rt-pcr methodand Western blot method. The expression of bcl-2 and McL-2 was deter- mined by Western - blot method. Results: Earthworm proteinhas inhibitory effect on the proliferation activity of HSF cells. Concentration of 0.625 mg/mL, 1.25 mg/mL sample group effect on the cellseparately after 48 hours, the apoptosis rate of the cell was 31.5% and 69.2%. method showed that concentration of 0.625 mg/mL, 1.25 mg/mL sample group was able to increase gene expression of Cas- pase-3, reduce gene expression of CyclinD1 and TGF -β1（P<0.01). There was a significant difference between the 0.3125 mg/mL sample group and the blank control group (P<0.05). Western - blot method showed that Each sample group could increase the protein expression of Caspase-3, reduce the protein expression of CyclinD1, and reduce the protein expression of TGF-beta 1 (P<0.01). The concentration of 0.625 mg/mL and 1.25 mg/mL samples was able to reduce the expression of bcl-2 protein (P<0.01). The protein expression of bcl-2 in the concentration of 0.3125 mg/mL sample group was significantly different than that in the blank control group (P<0.05). Conclusion: The mechanism of earthworm protein inducing cell death was related to that earthworm protein can cut Cyclin D1 gene and protein expres- sion, raise Caspase 3 gene and protein expression, cut TGF-β1 gene and protein expression, cut the Bcl-2 protein expression quantity. |
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