槟榔碱对口腔黏膜成纤维细胞病变的影响

Effect of arecoline on lesion of oral mucosa fibroblasts

目的分析槟榔碱对口腔粘膜成纤维细胞(OMFb)微丝骨架及胶原吞噬的影响。方法取正常人口腔黏膜,行组织块法培养,取第6代细胞行不同浓度槟榔碱(0μg/mL、10μg/mL、20μg/mL、30μg/mL、40μg/mL)干预,检测12h、24h、48h后细胞增殖情况;分析24h后胶原吞噬率变化情况;观察24h后细胞微丝骨架变化情况。结果槟榔碱干预12h后,不同槟榔碱浓度OMFb增殖情况差异无统计学意义(P>0.05);24h后,10μg/mL、20μg/mL浓度组OMFb增殖率上升,且20μg/mL浓度组增殖率>10μg/mL浓度组;30μg/mL、40μg/mL浓度组OMFb无增殖;48h后,各浓度槟榔碱均对OMFb增殖存在抑制效用。10μg/mL、20μg/mL浓度组OMFb胶原吞噬能力存在明显改善,而30μg/mL、40μg/mL浓度组胶原吞噬能力被抑制。此外,10μg/mL、20μg/mL浓度组其OMFb细胞微丝骨架表现为粗大束状排列,出现张力纤维,细胞极性增强,且10μg/mL浓度组变化情况高于20μg/mL浓度组;而40μg/mL浓度组其OMFb细胞微丝骨架明显减少,30μg/mL浓度组次之,细胞皮层出现大量染色聚集物质。结论低浓度槟榔碱在增强OMFb活性、胶原吞噬能力等方面具有显著促进作用,但高浓度则具有抑制作用。此外,低浓度的槟榔碱还可增强OMFb细胞微丝骨架聚合,而高浓度槟榔碱则可以抑制OMFb细胞微丝骨架聚合,这可能是口腔黏膜下纤维性变的主要发病机制之一。

Objective To analyze the effect of arecoline on mierofilament skeleton and phagocytosis of collagen of oral mucous fibroblasts （OMFb）. Methods Normal oral mucosa was taken and cultured, and the sixth generation of cell lines were intervened with arecoline at various concentrations （0, 10 μg/mL, 20 μg/mL, 30 μg/mL, 40 μg/mL）, cell proliferation was determined 12 h, 24 h, 48 h after intervention; The change of collagen phagoctosis rate and cell microfilament cytoskeleton 24 hours after were observed. Results Cell proliferation rate was stable 12 h after intervention with arecoline at various concentrations （P〉 0.05）; the cell proliferation 24 hours after intervention in 10 μg/mL group, and 20 μg/mL OMFb was increased, while no proliferation in 30μg/mL and 40μg/ml group OMFb was noticed; After 48 h, arecoline showed inhibition effect on proliferation of OMFb in 10 μg,/mL groups and the collagen phagocytosis ability of OMPh in 20 μg,/mL group OMFb was obviously improved, While collagen phagocytosis ability of OMPb in groups with concentration of 30 μg/mL and 40 μg/mL was suppressed. In addition, microfilament skeleton of OMFb in 10 μg/mL and 20 μg/mL group was characterized by thick beam pattern, tension of fiber, strengthened cell polarity. The number of microfilament cytoskeleton of OMFb in 40 μg/mL group was obviously reduced, followed by 30 μg/mL concentration group. A lot of dyeing elements gathered on cortex layer of OMFb. Conclusion Arecoline at low concentration was capable of enhancing the activity and collagen phagocytosis ability of OMFb, but arecoline exhibited inhibition capacity at high concentration. In addition, arecoline at low concentration can promote microfilament cytoskeleton aggregation of OMFb, while the microfilament cytoskeleton aggregation of OMFb was inhibited by arecoline at high concentration , this might be one of the main pathogenic mechanisms of lesions in oral sub mucous fibroblasts.