核心蛋白聚糖对翼状胬肉成纤维细胞增殖的影响

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,并对照观察丝裂霉素C(MMC)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HPF。(2)用0.01、0.1、1、5、10 mg/L DCN及 MMC分别作用于体外培养的HPF, 24 h、48 h、72 h后观察2种药物对HPF形态的改变,2种药物不同浓度分别作用12 h、24 h、48 h后 MTT法比较2种药物的作用效果,不同浓度的DCN作用48 h后免疫组织化学染色增殖细胞核抗原(PCNA)法检测细胞生长活性,流式细胞术测定细胞周期时相变化。结果: 10 mg/L DCN或1 mg/LMMC在12 h后均能显著抑制HPF的增殖(P<0.05),呈剂量和时间依赖性。1~10 mg/L DCN 作用48 h使G₀/G₁期细胞百分比上升(P<0.05),5~10 mg/LDCN 作用48 h使G₂/M期和S期百分比(G₂/M%+S%)显著下降(P<0.05),实验组和对照组均未检测到终末期凋亡细胞。1~10 mg/L DCN能浓度依赖性地抑制细胞表达PCNA(P<0.05)。结论: 核心蛋白聚糖能抑制HPF的增殖,阻滞细胞在DNA合成前期。

Inhibitory effect of decorin on human pterygium fibroblasts in vitro

AIM: To investigate the effect of decorin(DCN) on the proliferation and apoptosis of human pterygium fibroblasts(HPF) in vitro,and to compare the effect of mitomycin C(MMC) in order to search for a new method to prevent the recurrence after pterygium surgery.METHODS: Human pterygium fiborblasts were isolated from the caudomedial part of pterygium tissues in pterygium patients and then cultured in vitro using tissue inoculation method.The cells were treated with DCN and MMC at concentrations of 0.01,0.1,1,5 and 10 mg/L.The morphological alterations of HPF were observed after 24 h,48 h or 72 h of treatment.MTT method was used to assay the effects of the 2 drugs at different doses after 12 h,24 h and 48 h on the proliferation of the cells.The expression of proliferating cell nuclear antigen(PCNA) in each group treated with different doses of DCN was detected by the method of immunohistochemistry after 48 h.The cell cycle distribution was determined by flow cytometry analysis.RESULTS: After administration of 10 mg/L DCN or 1 mg/L MMC for 12 h,the proliferation of HPF was significantly inhibited by both drugs in a dose-and time-dependent manner(P<0.05).After treated with 1～10 mg/L DCN for 48 h,the percentage of HPF in G0/G1 phase was increased,while the percentage of HPF in G2/M phase and S phase(G2/M%+S%) was decreased after treated with 5～10mg/L DCN for 48 h(P<0.05).The late-apoptotic cells were not found in DCN group and MMC group.DCN dose-dependently inhibited the expression of PCNA in HPF(P<0.05).CONCLUSION: Decorin significantly inhibits the proliferation of HPF,and blocks the cells in G1 phase.