纹眼线诱导大鼠睑板腺功能障碍模型的建立与评估

Establishment and evaluation of induced model of meibomian gland dysfunction in rats through eyeliner tattoo

摘要:

目的：建立一种通过纹眼线的方式诱导大鼠睑板腺功能障碍的模型以及研究其可能机制。

方法：选取SD大鼠40只，随机取30只大鼠右眼纹眼线后为眼线组建立睑板腺功能障碍模型，余10只不采取任何处理为正常组，分别在建模后1、2、4 wk用裂隙灯观察两组大鼠角膜形态，并计算泪膜破裂时间(BUT)、泪液分泌量(SⅠt)、角膜荧光素钠染色评分和角膜不规则评分，眼表综合分析仪检查角膜Placido环，使用共聚焦显微镜观察两组大鼠角膜组织结构，建模4 wk后完成临床指标记录后，取眼球及上下眼睑组织行病理学检查，HE染色观察睑板腺结构，PAS染色观察结膜杯状细胞情况，ORO染色观察脂滴情况。

结果：裂隙灯检查结果显示，眼线组大鼠眼睑黑色色素在位，眼睑无变形及瘢痕，角膜上皮粗糙，荧光素染色阳性，呈点片状着染，随观察时间延长加重。与正常组大鼠比较，眼线组建模后1、2、4 wk BUT明显缩短、泪液分泌量明显下降，角膜荧光素钠染色评分及角膜不规则评分均明显升高(均P<0.01)。活体共聚焦显微镜检查结果显示，眼线组大鼠角膜上皮细胞减少，出现高亮异常的细胞，基质层可见炎症细胞浸润。ORO染色结果显示眼线组大鼠脂滴减少，随观察时间增加ORO染色呈现下降趋势。HE染色结果显示黑色色素阻塞眼线组大鼠睑板腺开口，随观察时间延长睑板腺腺泡密度呈现下降趋势。PAS染色结果显示眼线组大鼠PAS染色阳性细胞数量呈现下降趋势。

结论：纹眼线可诱导睑板腺功能障碍，所用色素颗粒造成的睑板腺开口堵塞，可能是导致睑板腺功能功能障碍的重要原因。

Abstract:

AIM:To establish a model of meibomian gland dysfunction in rats induced by eyeliner tattoo and investigate its potential mechanisms.

METHODS:A total of 40 SD rats were selected, with 30 randomly chosen to have eyeliner tattoo applied their right eyes and designated as the eyeliner group. The remaining 10 rats were not given any treatment and served as the normal group. The corneal morphology of both groups was observed using a slit lamp at 1, 2, and 4 wk after establishment, and the tear film break-up time(BUT), Schirmer I test(SIt), corneal fluorescein staining score, and corneal irregularity score were calculated. The corneal Placido rings were examined using an ocular surface analyzer, and the corneal tissue structures of both groups were observed under a confocal microscope. After 4 wk and completion of clinical indicator recording, the eyeballs and upper and lower eyelid tissues were taken for pathological examination. The meibomian gland structures were observed through HE staining, the conjunctival goblet cells were observed using PAS staining, and the lipid droplets were observed with ORO staining.

RESULTS:The slit lamp examination results showed that the eyeliner group rats exhibited in situ black pigmentation in the eyelids, with no eyelid deformation or scarring. The corneal epithelium was rough, with positive fluorescein staining, presenting as spotty staining that worsened over time. Compared with the normal group, the BUT was significantly shortened, tear secretion volume was significantly decreased, and the corneal fluorescein staining score and corneal irregularity score were significantly increased at 1, 2, and 4 wk after modeling in the eyeliner group(all P<0.01). The corneal confocal microscopy results showed a decrease in corneal epithelial cells in the eyeliner group, with the appearance of abnormally bright cells, and inflammatory cell infiltration visible in the stromal layer. The ORO staining results revealed a decrease in lipid droplets in the eyeliner group, showing a downward trend with increasing observation time. The HE staining results showed that pigment blocked the meibomian gland openings in the eyeliner group, and the density of meibomian gland acini showed a downward trend over time. The PAS staining results showed a decreasing trend in the number of PAS-positive cells in the eyeliner group.

CONCLUSION:Eyeliner tattoo can induce meibomian gland dysfunction, and the blockage of meibomian gland openings caused by the pigment particles used may be an important cause of meibomian gland dysfunction.