雷公藤多甙对体外培养的人翼状胬肉上皮鳞状化生的影响

Effects of tripterygium wilfordii multiglucoside on squamous metaplasia of human pterygium epithelium cultured in vitro

目的通过体外培养人翼状胬肉组织,探讨雷公藤多甙对人翼状胬肉上皮鳞状化生的影响。方法在体外含100μg·L-1雷公藤多甙(实验组)或不含雷公藤多甙(对照组)培养基浸没条件下体外培养人翼状胬肉组织7d和14d,观察形态学变化,同时应用免疫组织化学、免疫荧光组织化学、Westernblot方法测定胬肉组织上皮Pax6、K14和Ki67的表达变化,以检测翼状胬肉上皮细胞的增殖和分化状态。结果对照组培养7d后,翼状胬肉上皮层数增加,上皮变得不完整,但实验组未见明显变化。培养7d和14d时,两组上皮细胞层数比较差异均有统计学意义(t=1.778、2.393,均为P<0.05)。与培养前正常翼状胬肉组织相比,培养14d对照组Pax6在翼状胬肉上皮细胞核中的表达显著降低,K14未见明显改变,Ki67表达增加;而实验组Pax6在翼状胬肉上皮细胞核中的表达未见明显下降,Ki67、K14表达明显减少。实验组和对照组培养14d后,Pax6、Ki67、K14表达差异均有统计学意义(t=6.328、4.486、0.464,均为P<0.05)。结论体外翼状胬肉上皮鳞状化生的模型可通过培养基浸没法培养完成;雷公藤多甙可部分抑制翼状胬肉上皮的鳞状化生,这将为上皮鳞状化生性疾病的治疗开辟新的途径。

Objective To investigate the effect of tripterygium wilfordii multiglu- coside （TWM） on squamous metaplasia of human pterygium epithelium cultured in vitro. Methods Human pterygium tissues were cultured under submerge manner with （experiment group） or without 100 ~g ~ L-~ TWM （control group） for 7 days and 14 days, the changes of morphology were observed. The changes of Pax5, Ki57 and K14 ex- pression in pterygium epithelium were detected by immunohistochemistry, immunofluo- rescence staining and Western blot to investigate the proliferation and differentiation of pterygium epithelial cells. Results The pterygium epithelial layers increased, and the epithelium became disintact in control group after cultured for 7 days,but there was no obvious change in experimental group. There were statistical differences in the most pterygium epithelial layers after cultured for 7 days and 14 days between two groups （ t = 1. 778,2. 393, both P 〈 0.05 ）. Compared with the normal pterygium tissue, the ex- pression of Pax5 in pterygium epithelial cell nuclei at 14 days in control group after cul- ture was obviously decreased, K14 had no obviously changes, and Ki57 increased; Pax5 in experiment group was not decreased, and Ki57 and K14 obviously changed. There were statistical differences in Pax6, Ki57 and K14 expression at 14 days after culture be- tween control group and experiment group （ t = 5. 328,4. 485,0. 454, both P 〈 0.05 ）. Conclusion The model of squamous metaplasia of pterygium epithelium cultured in vitro can be established by medium submersion. TWM can partly inhibit the squamous metaplasia of pterygium epithelium, which may shed new light on treatment for diseases with epithelial squamous metaplasia.