饰胶蛋白聚糖对翼状胬肉成纤维细胞增殖的影响

发布时间：2019-01-07 22:22

【摘要】： [目的]观察核心蛋白聚糖DCN对体外培养人翼状胬肉成纤维细胞(HPF)增殖的影响,并对照丝裂霉素对HPF的影响,寻找辅助治疗和预防翼状胬肉复发的新途径。 [方法] 1.取患者的翼状胬肉体部组织,采用组织块贴壁培养法原代培养人翼状胬肉成纤维细胞。 2.用0.01、0.1、1、5、10mg/L的DCN及MMC分别作用于体外培养的HPF，24h、48h、72h后观察两种药物对HPF细胞形态的改变,两种药物不同浓度分别作用12h、24h、48h后MTT法比较两种药物的作用效果,不同浓度的DCN作用48小时后免疫组织化学染色增生细胞核抗原(PCNA)法检测细胞生长活性,流式细胞仪测定细胞周期时相变化。 [结果]MTT法检测10 mg/L的DCN,1mg/L的MMC在12h后均能显著抑制HPF的增殖(P0.05),呈剂量和时间依赖性。DCN作用48h后,流式细胞仪检测结果发现,1-10mg/L的DCN G0/G1期细胞百分比上升(P0.05),5-10mg/LS期细胞百分比及增值率(G2/M%+S%)显著下降(P0.05),实验组和对照组均未检测到终末期凋亡细胞。当DCN的浓度在1-10 mg/L范围内能浓度依赖性地抑制细胞表达PCNA(P0.05)。 [结论]饰胶蛋白聚糖能抑制HPF的增殖,流式细胞仪检测显示DCN阻滞细胞在DNA合成前期。
[Abstract]:[objective] to observe the effect of core proteoglycan DCN on the proliferation of human pterygium fibroblasts (HPF) in vitro, and to find a new way to treat and prevent the recurrence of pterygium by comparing the effect of mitomycin on HPF. [methods] 1. Human pterygium fibroblasts were cultured by tissue mass adherent culture. 2. 10 mg / L DCN and MMC were used to observe the morphological changes of HPF cells in vitro after treated with 0. 01 mg / L DCN and 10 mg / L MMC. The two drugs were exposed to different concentrations for 12 h or 24 h, respectively. After 48 hours, the effects of the two drugs were compared by MTT method. The cell growth activity was detected by immunohistochemical staining of proliferating cell nuclear antigen (PCNA) after 48 hours of exposure to different concentrations of DCN, and the phase changes of cell cycle were measured by flow cytometry. [results] the MMC of 10 mg/L DCN,1mg/L detected by MTT could significantly inhibit the proliferation of HPF (P0.05) in a dose-and time-dependent manner after 12 h. The results of flow cytometry showed that after 48 h of DCN treatment, the proliferation of HPF was significantly inhibited. The percentage of DCN G0/G1 phase cells in 1-10mg/L increased (P0.05), while the percentage of 5-10mg/LS phase cells and the proliferation rate (G2 / M% S%) decreased significantly (P0.05). No end-stage apoptotic cells were detected in the experimental group and the control group. When the concentration of DCN was in the range of 1-10 mg/L, the expression of PCNA was inhibited in a concentration-dependent manner (P0.05). [conclusion] Glucoglycan can inhibit the proliferation of HPF. Flow cytometry showed that DCN blocked the cells in the early stage of DNA synthesis.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R777.33

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