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目的 观察蛇毒制剂对体外培养的兔结膜下成纤维细胞与胶原黏附及细胞移行和增殖的影响。方法 将培养的第 3～ 5代兔结膜下成纤维细胞于不同浓度的蛇毒制剂孵育后 ,接种于涂有鼠尾胶原的 2 4孔板中 ,去除未黏附的细胞 ,用目镜网格器计数法和四甲基偶氮唑盐法记录贴壁细胞量。在第 3代生长融合细胞的培养板上划线 ,造成无细胞的裸露区 ,在不同浓度的蛇毒制剂下孵育 ,用目镜网格器每 6h观察记录移行至裸露区的细胞数。第 3代兔结膜下成纤维细胞接种于 2 4孔板与不同浓度的蛇毒制剂共育 2 4、48h后 ,用四甲基偶氮唑盐法记录各孔细胞数量的吸光度 (A)值。结果蛇毒制剂抑制结膜下成纤维细胞黏附呈剂量依赖性。抑制成纤维细胞黏附的半数有效量 (ID50 )为1 0× 10 -3 U/ml。 5 0× 10 -3 U/ml的蛇毒制剂能够抑制结膜下成纤维细胞的移行。 1 0～ 5 0× 10 -3U/ml蛇毒制剂不能显著地抑制结膜下成纤维细胞增殖。结论 蛇毒制剂浓度 >1 0× 10 -3 U/ml,能够显著地抑制兔结膜下成纤维细胞与胶原的黏附 ,5 0× 10 -3 U/ml的蛇毒制剂能显著抑制细胞移行 ,但不影响细胞自发性增殖
Objective To observe the effect of snake venom preparation on the adhesion and cell migration and proliferation of rabbit subconjunctival fibroblasts cultured in vitro. Methods The cultured 3rd to 5th generation rabbit subconjunctival fibroblasts were incubated in different concentrations of snake venom preparations and inoculated into 24-well plates coated with rat tail collagen to remove non-adherent cells and counted with an eyepiece grid Adherent cells were recorded by the method of MTT assay. The 3rd generation of fused cells were streaked on the plate, resulting in a cell free zone exposed to different concentrations of snake venom preparations. The number of cells migrated to the bare zone was recorded every 6 h with an eyepiece grid. The third generation of rabbit subconjunctival fibroblasts were seeded on 24-well plates with different concentrations of snake venom for 2, 48 hours. The absorbance (A) value of each well was recorded by MTT method. Results The snake venom preparation inhibited the adhesion of subconjunctival fibroblasts in a dose - dependent manner. The median effective dose (ID50) for inhibiting fibroblast adhesion was 10 × 10 -3 U / ml. The snake venom preparation at 5 × 10 -3 U / ml can inhibit the transplantion of subconjunctival fibroblasts. 10 ~ 50 × 10 -3 U / ml snake venom preparation can not significantly inhibit the proliferation of sub-epithelial fibroblasts. Conclusion The concentration of snake venom> 10 × 10 -3 U / ml can significantly inhibit the adhesion of rabbit subconjunctival fibroblasts to collagen. The snake venom preparation with the dosage of 50 × 10 -3 U / ml can significantly inhibit the cell migration but not Affect the spontaneous proliferation of cells