目的：探讨木犀草素(luteolin)对青光眼小梁切除术术后滤过区组织纤维化的影响及其与Wnt3a/β-catenin通路的关系。

方法：构建兔青光眼小梁切除术后纤维化模型，随机分为假手术组、模型组、luteolin-L组、luteolin-H组。动态监测各组术后眼压波动； 借助裂隙灯显微镜观察术后滤过区结膜组织的形态、结膜愈合程度、角膜完整性以及前房炎症反应； HE染色评估滤过区结膜上皮完整性、上皮下纤维组织增生程度及炎性细胞浸润情况； Masson三色染色分析胶原纤维沉积情况进而直观反映组织纤维化的程度； 免疫组织化学法染色观察α-SMA的表达情况。通过RT-qPCR与Western blot技术，分别定量检测术区结膜组织中Wnt3a、Wnt7a、Wnt2b、β-catenin、LC3-Ⅰ、LC3-Ⅱ、p62-SQSTM1的mRNA与蛋白表达水平。

结果：术后动态监测结果显示，与模型组相比，luteolin-L组和luteolin-H组整体眼压控制效果更优，且luteolin-H组显著优于luteolin-L组(均P<0.05)； 裂隙灯观察证实，luteolin可有效改善术后结膜损伤，减轻前房炎症反应，维持滤过区组织结构稳定。HE和Masson染色显示，与模型组相比，luteolin干预后结膜上皮损伤减轻，炎性细胞浸润减少，成纤维细胞增殖、新生血管生成及胶原过度沉积受到明显抑制，滤过泡瘢痕化进程显著延缓，且luteolin-H组影响程度较luteolin-L组更高； 免疫组织化学染色显示，luteolin干预可降低α-SMA的阳性表达率； 分子检测结果显示，luteolin-H组术区Wnt3a、β-catenin、LC3-Ⅱ/LC3-Ⅰ表达程度低于模型组和luteolin-L组，p62-SQSTM1表达程度最高(均P<0.05)，Wnt2b、Wnt7a在各组间表达程度无明显差异，说明luteolin能够特异性调控Wnt3a通路且剂量依赖性抑制青光眼小梁切除术后Wnt3a、β-catenin通路激活，下调LC3-Ⅱ/LC3-Ⅰ表达、上调p62-SQSTM1水平，抑制过度自噬活化。

结论：luteolin能明显抑制小梁切除术术后滤过泡组织纤维化，可能是通过Wnt3a/β-catenin通路来减弱纤维化程度。

METHODS: Rabbits were selected to establish a fibrosis model after glaucoma trabeculectomy, and randomly divided into the sham operation group, model group, luteolin-L group and luteolin-H group. Dynamic monitoring of postoperative intraocular pressure was performed in all groups. Slit-lamp biomicroscopy was used to observe postoperative conjunctival morphological changes, conjunctival healing degree, corneal integrity, and anterior chamber inflammatory responses in the filtering bleb area. HE staining was performed to evaluate conjunctival epithelial integrity, subepithelial fibrous proliferation, and inflammatory cell infiltration in the filtering tissues. Masson's trichrome staining was applied to assess collagen deposition and the degree of tissue fibrosis. The expression of α-SMA was observed by immunohistochemical staining. The mRNA and protein expression levels of Wnt3a, Wnt7a, Wnt2b, β-catenin, LC3-I, LC3-II and p62-SQSTM1 in conjunctival tissues of the operative area were quantitatively detected by RT-

qPCR and Western blot, respectively.

RESULTS:Postoperative dynamic monitoring showed that the luteolin-H group achieved significantly better intraocular pressure control than the luteolin-L group compared with the model group(all P<0.05). Slit lamp observations confirmed that luteolin effectively ameliorated postoperative conjunctival injury, alleviated anterior chamber inflammatory response, and maintained the structural stability of the filtering area. HE and Masson staining revealed that luteolin intervention reduced conjunctival epithelial damage and inflammatory cell infiltration, markedly inhibited fibroblast proliferation, neovascularization and excessive collagen deposition, and significantly delayed the progression of filtering bleb scarring, with stronger effects in the luteolin-H group than luteolin-L group compared with the model group. Immunohistochemical staining demonstrated that luteolin intervention decreased the positive expression rate of α-SMA. Molecular assays indicated that the luteolin-H group had lower expression levels of Wnt3a, β-catenin and LC3-II/LC3-I, and the highest p62-SQSTM1 level in the operative site relative to the model group and luteolin-L group(all PSymbol|@@0.05). No significant intergroup differences were observed in Wnt2b and Wnt7a expression. These findings suggested that luteolin could specifically regulated the Wnt3a pathway and dose-dependently suppressed the activation of the Wnt3a/β-catenin pathway after glaucoma trabeculectomy, downregulated LC3-II/LC3-I expression, upregulated p62-SQSTM1 levels, and inhibited excessive autophagy activation.

CONCLUSION: Luteolin can significantly inhibit filtering bleb fibrosis after trabeculectomy, which may reduce the degree of fibrosis via the Wnt3a/β-catenin pathway.